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A Plant Dj-1 Homolog Is Essential For Arabidopsis Thaliana Chloroplast Development, Jiusheng Lin, Tara J. Nazarenus, Jeanine L. Frey, Xinwen Liang, Mark A. Wilson, Julie M. Stone

Center for Plant Science Innovation: Faculty Publications

Protein superfamilies can exhibit considerable diversification of function among their members in various organisms. The DJ-1 superfamily is composed of proteins that are principally involved in stress response and are widely distributed in all kingdoms of life. The model flowering plant Arabidopsis thaliana contains three close homologs of animal DJ-1, all of which are tandem duplications of the DJ-1 domain. Consequently, the plant DJ-1 homologs are likely pseudo-dimeric proteins composed of a single polypeptide chain. We report that one A. thaliana DJ-1 homolog (AtDJ1C) is the first DJ-1 homolog in any organism that is required for viability. Homozygous disruption of …

Two Distinct Roles Of Arabidopsis Homolog Of Trithorax1 (Atx1) At Promoters And Within Transcribed Regions Of Atx1-Regulated Genes, Yong Ding, Zoya Avramova, Michael E. Fromm

Center for Plant Science Innovation: Faculty Publications

The Arabidopsis thaliana trithorax-like protein, ATX1, shares common structural domains, has similar histone methyltransferase (HMT) activity, and belongs in the same phylogenetic subgroup as its animal counterparts. Most of our knowledge of the role of HMTs in trimethylating lysine 4 of histone H3 (H3K4me3) in transcriptional regulation comes from studies of yeast and mammalian homologs. Little is known about the mechanism by which ATX1, or any other HMT of plant origin, affects transcription. Here, we provide insights into how ATX1 influences transcription at regulated genes, playing two distinct roles. At promoters, ATX1 is required for TATA binding protein (TBP) and …

Structure Function Analysis Of An Adp-Ribosyltransferase Type Iii Effector And Its Rna-Binding Target In Plant Immunity, Byeong-Ryool Jeong, Yan Lin, Anna Joe, Ming Guo, Christin Korneli, Huirong Yang, Ping Wang, Min Yu, Ronald Cerny, Dorothee Staiger, James R. Alfano, Yanhui Xu

Center for Plant Science Innovation: Faculty Publications

Background: HopU1 ADP-ribosylates GRP7, suppressing plant immunity.

Results: The HopU1 structure has two novel loops required for GRP7 recognition, and HopU1 ribosylates GRP7 at an arginine in position 49 disrupting its function.

Conclusion: HopU1 targets a conserved arginine in GRP7, disabling its ability to bind immunity-related RNA.

Significance: The mechanistic details of how HopU1 recognizes its substrate reveal how HopU1 contributes to pathogenesis.

Functional Hak/Kup/Kt-Like Potassium Transporter Encoded By Chlorella Viruses, Timo Greiner, José Ramos, Maria C. Alvarez, James Gurnon, Ming Kang, James L. Van Etten, Anna Moroni, Gerhard Thiel

James Van Etten Publications

Chlorella viruses are a source of interesting membrane transport proteins. Here we examine a putative K+ transporter encoded by virus FR483 and related chlorella viruses. The protein shares sequence and structural features with HAK/KUP/KT-like K+ transporters from plants, bacteria and fungi. Yeast complementation assays and Rb+ uptake experiments show that the viral protein, termed HAKCV (high-affinity K+ transporter of chlorella virus), is functional, with transport characteristics that are similar to those of known K+ transporters. Expression studies revealed that the protein is expressed as an early gene during viral replication, and proteomics data indicate that it is not packaged in …

Characterization Of Viral And Host Proteins And Rna Elements In Tombusvirus Replication, Kunj Bihari Pathak

Theses and Dissertations--Plant Pathology

Two thirds of plant viruses are positive-strand RNA viruses including the family Tombusviridae. One of the best-studied members of this family is Tomato bushy stunt virus (TBSV). Like many other viruses, TBSV has much fewer genes when compared to its hosts’ genome. Nevertheless, TBSV utilizes its genome very judiciously. To compensate for a lack of many proteins of its own, it codes for multi-functional replication protein p33 and also co-opts host factors to facilitate its replication.

By using recombinant replication proteins p33 and p92 containing single amino acid changes in protein-protein interaction domains (S1 and S2), I demonstrated that the …

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. …

Giant Viruses, James L. Van Etten

James Van Etten Publications

The common view of viruses, mostly true, is of tiny burglars that sneak into cells, grab the biosynthetic controls and compel the cell to make huge numbers of progeny that break out of the cell and keep the replication cycle going. Viruses are supposed to be diminutive even compared to cells that are just a micrometer (1,000 nanometers) in diameter. They are supposed to travel light, making do with just a few well-adapted genes.

In 1992, a new microorganism was isolated from a power-plant cooling tower in Bradford, England, where Timothy Robotham, a microbiologist at Leeds Public Health Laboratory, was …

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 …

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 …

Comparison Of Plant‐Adapted Rhabdovirus Protein Localization And Interactions, Kathleen Marie Martin

University of Kentucky Doctoral Dissertations

Sonchus yellow net virus (SYNV), Potato yellow dwarf virus (PYDV) and Lettuce Necrotic yellows virus (LNYV) are members of the Rhabdoviridae family that infect plants. SYNV and PYDV are Nucleorhabdoviruses that replicate in the nuclei of infected cells and LNYV is a Cytorhabdovirus that replicates in the cytoplasm. LNYV and SYNV share a similar genome organization with a gene order of Nucleoprotein (N), Phosphoprotein (P), putative movement protein (Mv), Matrix protein (M), Glycoprotein (G) and Polymerase protein (L). PYDV contains an additional predicted gene between N and P, denoted as X, that has an unknown function. In order to gain …

Abundant Degenerate Miniature Inverted-Repeat Transposable Elements In Genomes Of Epichloid Fungal Endophytes Of Grasses, Damien J Fleetwood, Anar K Khan, Richard D Johnson, Carolyn A Young, Shipra Mittal, Ruth E Wrenn, Uljana Hesse, Simon J Foster, Christopher L. Schardl, Barry Scott

Plant Pathology Faculty Publications

Miniature inverted-repeat transposable elements (MITEs) are abundant repeat elements in plant and animal genomes; however, there are few analyses of these elements in fungal genomes. Analysis of the draft genome sequence of the fungal endophyte Epichloë festucae revealed 13 MITE families that make up almost 1% of the E. festucae genome, and relics of putative autonomous parent elements were identified for three families. Sequence and DNA hybridization analyses suggest that at least some of the MITEs identified in the study were active early in the evolution of Epichloë but are not found in closely related genera. Analysis of MITE integration …

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 …

Regalia-Induced Resistance In Powdery Mildew-Infected Grapevine Plants Correlates With The Induction Of Chalcone Synthase, Thomas Crottogini

Horticulture and Crop Science

No abstract provided.

An Nadph-Dependent Genetic Switch Regulates Plant Infection By The Rice Blast Fungus, Richard A. Wilson, Robert P. Gibson, Cristian Quispe, Jennifer A. Littlechild, Nicholas J. Talbot

Fungal Molecular Plant-Microbe Interactions

To cause rice blast disease, the fungus Magnaporthe oryzae breaches the tough outer cuticle of the rice leaf by using specialized infection structures called appressoria. These cells allow the fungus to invade the host plant and proliferate rapidly within leaf tissue. Here, we show that a unique NADPH-dependent genetic switch regulates plant infection in response to the changing nutritional and redox conditions encountered by the pathogen. The biosynthetic enzyme trehalose-6-phosphate synthase (Tps1) integrates control of glucose-6-phosphate metabolism and nitrogen source utilization by regulating the oxidative pentose phosphate pathway, the generation of NADPH, and the activity of nitrate reductase. We report …

Population Variability Of Rotylenchulus Reniformis In Cotton Agroecosystems, Megan Leach

All Dissertations

Rotylenchulus reniformis, reniform nematode, is a highly variable species and an economically important pest in many cotton fields across the southeast. Rotation to resistant or poor host crops is a prescribed method for management of reniform nematode. An increase in the incidence and prevalence of the nematode in the United States has been reported over the last two decades. However it is not clear whether the observed increase is related to the emergence of novel populations that are more aggressive or have a higher fitness or shifts in host availability or susceptibility. The objectives of this research were to determine …

Analysis Of The Response Of Medicago Truncatula Calcium Oxalate Mutants To Abiotic Stress, Wayra Gabriela Navia-Gine

Graduate Theses and Dissertations

Calcium oxalate crystals are found in most plant species. In Medicago truncatula wild-type A17, crystals accumulate in leaves along the secondary veins and the only role attributed to them so far is defense against chewing insects. Calcium oxalate deficient (cod) mutants were isolated in M. truncatula; the cod mutants include cod5, which completely lack crystals in the leaves and cod6 that accumulates fewer and smaller crystals compared to A17. We analyzed gene expression in the cod mutants and A17 using GeneChip® Medicago Genome Arrays and found important differences in transcriptome between the three genotypes. In particular, we found a gene …

Fusarium Head Blight: Winter Wheat Cultivar Responses And Characterization Of Pathogen Isolates, John Fredy Hernandez Nopsa

Department of Agronomy and Horticulture: Dissertations, Theses, and Student Research

Fusarium Head Blight (FHB) is a destructive disease of wheat (Triticum aestivum L). FHB reduces yield and grain quality and causes accumulation of the mycotoxin deoxynivalenol (DON) in grain. Cultivar resistance is one of the most effective management strategies for FHB. Experiments were conducted to 1) identify winter wheat cultivars with resistance to FHB and DON accumulation, 2) determine the effect of winter wheat cultivar on the relationship between FHB and DON concentration, and 3) identify the major species of Fusarium causing FHB in Nebraska and characterize its isolates. Differences (P ≤ 0.05) were detected among cultivars in …

Identification Of Sources Of Rhizoctonia Root Rot Resistance In Common Bean And Mapping A New Source Of Bean Rust Resistance From The Tertiary Gene Pool Of Common Bean, Pamela A. Peña-Perdomo

Department of Agronomy and Horticulture: Dissertations, Theses, and Student Research

An efficient screening method was developed and used to identify bean lines resistant to Rhizoctonia Root Rot caused by Rhizoctonia solani Kuhn. Two sets of 163 and 111 lines previously evaluated for drought tolerance at Mitchell, NE and Isabela, PR were evaluated for Rhizoctonia Root Rot resistance under greenhouse conditions. This root rot data was also correlated with yield under drought stress and non stress conditions. In the first set of lines the rhizoctonia mean score ranged from 1.7 to 3.9; and in the second set the rhizoctonia mean score was between 2.6 and 5.7. There was no significant correlation …

The Combined Effect Of Environmental And Host Factors On The Emergence Of Viral Rna Recombinants, Hannah M. Jaag, Peter D. Nagy

Plant Pathology Faculty Publications

Viruses are masters of evolution due to high frequency mutations and genetic recombination. In spite of the significance of viral RNA recombination that promotes the emergence of drug-resistant virus strains, the role of host and environmental factors in RNA recombination is poorly understood. Here we report that the host Met22p/Hal2p bisphosphate-3'-nucleotidase regulates the frequency of viral RNA recombination and the efficiency of viral replication. Based on Tomato bushy stunt virus (TBSV) and yeast as a model host, we demonstrate that deletion of MET22 in yeast or knockdown of AHL, SAL1 and FRY1 nucleotidases/phosphatases in plants leads to increased TBSV recombination …

Plant Immunity Directly Or Indirectly Restricts The Injection Of Type Iii Effectors By The Pseudomonas Syringae Type Iii Secretion System, Emerson Crabill, Anna Joe, Anna Block, Jennifer M. Van Rooyen, James R. Alfano

Center for Plant Science Innovation: Faculty Publications

Plants perceive microorganisms by recognizing microbial molecules known as pathogen-associated molecular patterns (PAMPs) inducing PAMP-triggered immunity (PTI) or by recognizing pathogen effectors inducing effector-triggered immunity (ETI). The hypersensitive response (HR), a programmed cell death response associated with ETI, is known to be inhibited by PTI. Here, we show that PTI-induced HR inhibition is due to direct or indirect restriction of the type III protein secretion system’s (T3SS) ability to inject type III effectors (T3Es). We found that the Pseudomonas syringae T3SS was restricted in its ability to inject a T3E-adenylate cyclase (CyaA) injection reporter into PTI-induced tobacco (Nicotiana tabacum) cells. …

The Chlorella Variabilis Nc64a Genome Reveals Adaptation To Photosymbiosis, Coevolution With Viruses, And Cryptic Sex, Guillaume Blanc, Garry Duncan, Irina Agarkova, Mark Borodovsky, James Gurnon, Alan Kuo, Erika Lindquist, Susan Lucas, Jasmyn Pangilinan, Juergen Polle, Asaf Salamov, Astrid Terry, Takashi Yamada, David D. Dunigan, Igor V. Grigoriev, Jean-Michel Claverie, James L. Van Etten

Department of Plant Pathology: Faculty Publications

Chlorella variabilis NC64A, a unicellular photosynthetic green alga (Trebouxiophyceae), is an intracellular photobiont of Paramecium bursaria and a model system for studying virus/algal interactions. We sequenced its 46-Mb nuclear genome, revealing an expansion of protein families that could have participated in adaptation to symbiosis. NC64A exhibits variations in GC content across its genome that correlate with global expression level, average intron size, and codon usage bias. Although Chlorella species have been assumed to be asexual and nonmotile, the NC64A genome encodes all the known meiosis-specific proteins and a subset of proteins found in flagella. We hypothesize that Chlorella might have …

Increased Plant Uptake Of Nitrogen From 15N-Depleted Fertilizer Using Plant Growth-Promoting Rhizobacteria, A. O. Adesemoye, H. A. Torbert, J. W. Kloepper

Department of Plant Pathology: Faculty Publications

Harmful environmental effects resulting from fertilizer use have spurred research into integrated nutrient management strategies which can include the use of specific microorganisms to enhance nutrient use efficiency by plants. Some strains of plant growth-promoting rhizobacteria (PGPR) have been reported to enhance nutrient uptake by plants, but no studies with PGPR have used ¹⁵N isotope techniques to prove that the increased N in plant tissues came from the N applied as fertilizer. The current study was conducted to demonstrate that a model PGPR system can enhance plant uptake of fertilizer N applied to the soil using different rates of …