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Dna Barcoding Evidence For The North American Presence Of Alfalfa Cyst Nematode, Heterodera Medicaginis, Tom Powers, Andrea Skantar, Timothy Harris, Rebecca Higgins, Peter Mullin, Saad Hafez, Zafar Handoo, Tim Todd, Kirsten S. Powers

Department of Plant Pathology: Faculty Publications

Specimens of Heterodera have been collected from alfalfa fields in Kearny County, Kansas and Carbon County, Montana. DNA barcoding with the COI mitochondrial gene indicate that the species is not Heterodera glycines, soybean cyst nematode, H. schachtii, sugar beet cyst nematode, or H. trifolii, clover cyst nematode. Maximum likelihood phylogenetic trees show that the alfalfa specimens form a sister clade most closely related to H. glycines, with a 4.7% mean pairwise sequence divergence across the 862 nucleotides of the COI marker. Morphological analyses of juveniles and cysts conform to the measurements of H. medicaginis, the alfalfa cyst nematode originally described …

Clarireedia: A New Fungal Genus Comprising Four Pathogenic Species Responsible For Dollar Spot Disease Of Turfgrass, Catalina Salgado-Salazar, Lisa A. Beirn, Adnan Ismaiel, Michael J. Boehm, Ignazio Carbone, Alexander I. Putman, Lane P. Tredway, Bruce B. Clarke, Jo Anne Crouch

Department of Plant Pathology: Faculty Publications

Dollar spot is one of the most destructive and economically important fungal diseases of amenity turfgrasses. The causal agent was first described in 1937 as the ascomycete Sclerotinia homoeocarpa. However, the genus-level taxonomic placement of this fungus has been the subject of an ongoing debate for over 75 y. Existing morphological and rDNA sequence evidence indicates that this organism is more appropriately placed in the family Rutstroemiaceae rather than the Sclerotiniaceae. Here we use DNA sequence data from samples of the dollar spot fungus and other members of the Rutstroemiaceae (e.g. Rutstroemia, Lanzia, Lambertella) collected throughout the world to determine …

Reactive Oxygen Species Metabolism And Plant-Fungal Interactions, Lauren M. Segal, Richard A. Wilson

Department of Plant Pathology: Faculty Publications

Fungal interactions with plants can involve specific morphogenetic developments to access host cells, the suppression of plant defenses, and the establishment of a feeding lifestyle that nourishes the colonizer often—but not always—at the expense of the host. Reactive oxygen species (ROS) metabolism is central to the infection process, and the stage-specific production and/or neutralization of ROS is critical to the success of the colonization process. ROS metabolism during infection is dynamic—sometimes seemingly contradictory—and involves endogenous and exogenous sources. Yet, intriguingly, molecular decision-making involved in the spatio-temporal control of ROS metabolism is largely unknown. When also considering that ROS demands are …

Tomato Spotted Wilt Virus Nss Protein Supports Infection And Systemic Movement Of A Potyvirus And Is A Symptom Determinant, Hernan Garcia-Ruiz, Sergio M. Gabriel Peralta, Patricia A. Harte-Maxwell

Department of Plant Pathology: Faculty Publications

Plant viruses are inducers and targets of antiviral RNA silencing. To condition susceptibility, most plant viruses encode silencing suppressor proteins that interfere with antiviral RNA silencing. The NSs protein is an RNA silencing suppressor in orthotospoviruses, such as the tomato spotted wilt virus (TSWV). The mechanism of RNA silencing suppression by NSs and its role in virus infection and movement are poorly understood. Here, we cloned and tagged TSWV NSs and expressed it from a GFP-tagged turnip mosaic virus (TuMV-GFP) carrying either a wild-type or suppressor-deficient (AS9) helper component proteinase (HC-Pro). When expressed in cis, NSs restored pathogenicity and promoted …

Pushing The Resolution Limit By Correcting The Ewald Sphere Effect In Single-Particle Cryo-Em Reconstructions, Dongjie Zhu, Xiangxi Wang, Qianglin Fang, James L. Van Etten, Michael G. Rossmann, Zihe Rao, Xinzheng Zhang

Department of Plant Pathology: Faculty Publications

The Ewald sphere effect is generally neglected when using the Central Projection Theorem for cryo electron microscopy single-particle reconstructions. This can reduce the resolution of a reconstruction. Here we estimate the attainable resolution and report a “block-based” reconstruction method for extending the resolution limit. We find the Ewald sphere effect limits the resolution of large objects, especially large viruses. After processing two real datasets of large viruses, we show that our procedure can extend the resolution for both datasets and can accommodate the flexibility associated with large protein complexes.

Metagenomic Analysis Of Viruses Associated With Maize Lethal Necrosis In Kenya, Mwathi Jane Wamaitha, Deepti Nigam, Solomon Maina, Francesca Stomeo, Anne Wangai, Joyce Njoki Njuguna, Timothy A. Holton, Bramwel W. Wanjala, Mark Wamalwa, Tanui Lucas, Appolinaire Djikeng, Hernan Garcia Ruiz

Department of Plant Pathology: Faculty Publications

Background: Maize lethal necrosis is caused by a synergistic co-infection of Maize chlorotic mottle virus (MCMV) and a specific member of the Potyviridae, such as Sugarcane mosaic virus (SCMV), Wheat streak mosaic virus (WSMV) or Johnson grass mosaic virus (JGMV). Typical maize lethal necrosis symptoms include severe yellowing and leaf drying from the edges. In Kenya, we detected plants showing typical and atypical symptoms. Both groups of plants often tested negative for SCMV by ELISA. Methods: We used next-generation sequencing to identify viruses associated to maize lethal necrosis in Kenya through a metagenomics analysis. Symptomatic and asymptomatic leaf samples …

Genome-Wide Analysis Of Genes Encoding Core Components Of The Ubiquitin System In Soybean (Glycine Max) Reveals A Potential Role For Ubiquitination In Host Immunity Against Soybean Cyst Nematode, Chunyu Zhang, Li Song, Mani Kant Choudhary, Bangjun Zhou, Guangchao Sun, Kyle C. Broderick, Loren Giesler, Lirong Zeng

Department of Plant Pathology: Faculty Publications

Background: Ubiquitination is a major post-translational protein modification that regulates essentially all cellular and physiological pathways in eukaryotes. The ubiquitination process typically involves three distinct classes of enzymes, ubiquitin-activating enzyme (E1), ubiquitin-conjugating enzyme (E2) and ubiquitin ligase (E3). To date, a comprehensive identification and analysis of core components comprising of the whole soybean (Glycine max) ubiquitin system (UBS) has not been reported.

Results: We performed a systematic, genome-wide analysis of genes that encode core members of the soybean UBS in this study. A total of 1431 genes were identified with high confidence to encode putative soybean UBS components, …

Evaluation Of Bacillus Strains For Plant Growth Promotion And Predictability Of Efficacy By In Vitro Physiological Traits, Rufus J. Akinrinlola, Gary Y. Yuen, Rhae A. Drijber, Anthony O. Adesemoye

Department of Plant Pathology: Faculty Publications

Bacilli are commonly used as plant growth-promoting agents but can be limited in effectiveness to certain crop and soil environments. The objectives of this study were to (1) identify Bacillus strains that can be consistent in promoting the growth of corn, wheat, and soybean and (2) determine whether physiological traits expressed in vitro can be predictive of growth promotion efficacy/consistency and be used for selecting effective strains. Twelve Bacillus strains isolated from wheat rhizospheres were evaluated in greenhouse pot tests with nonsterile soil for their effects on the growth of corn, soybean, and wheat. The strains also were assessed in …

The Tomato U-Box Type E3 Ligase Pub13 Acts With Group Iii Ubiquitin E2 Enzymes To Modulate Fls2-Mediated Immune Signaling, Bangjun Zhou, Lirong Zeng

Department of Plant Pathology: Faculty Publications

In Arabidopsis and rice, the ubiquitin ligase PUB13-mediated protein degradation plays a significant role in plant pattern-triggered immunity (PTI) and flowering time control. The Arabidopsis PUB13 has been shown to attenuate the pattern recognition receptor FLS2-mediated immune signaling by ubiquitinating FLS2 and consequently promoting its degradation by the 26S proteasome. Nevertheless, the cognate ubiquitin-conjugating enzymes (E2) with which PUB13 acts to modulate FLS2-mediated PTI are unknown. To address this question, we investigate here the tomato (Solanum lycopersicum) homolog of PUB13, SlPUB13 by utilizing the recently characterized complete set of tomato E2s. Of the 13 groups of tomato E2s, …

Susceptibility Genes To Plant Viruses, Hernan Garcia Ruiz

Department of Plant Pathology: Faculty Publications

Plant viruses use cellular factors and resources to replicate and move. Plants respond to viral infection by several mechanisms, including innate immunity, autophagy, and gene silencing, that viruses must evade or suppress. Thus, the establishment of infection is genetically determined by the availability of host factors necessary for virus replication and movement and by the balance between plant defense and viral suppression of defense responses. Host factors may have antiviral or proviral activities. Proviral factors condition susceptibility to viruses by participating in processes essential to the virus. Here, we review current advances in the identification and characterization of host factors …

Elucidating The Role Of Highly Homologous Nicotiana Benthamiana Ubiquitin E2 Gene Family Members In Plant Immunity Through An Improved Virus‑Induced Gene Silencing Approach, Bangjun Zhou, Lirong Zeng

Department of Plant Pathology: Faculty Publications

Background: Virus-induced gene silencing (VIGS) has been used in many plant species as an attractive post transcriptional gene silencing (PTGS) method for studying gene function either individually or at large-scale in a high-throughput manner. However, the specificity and efficiency for knocking down members of a highly homologous gene family have remained to date a significant challenge in VIGS due to silencing of off-targets. Results: Here we present an improved method for the selection and evaluation of gene fragments used for VIGS to specifically and efficiently knock down members of a highly homologous gene family. Using this method, we knocked down …

Proteomic Analysis Of Ubiquitinated Proteins In Rice (Oryza Sativa) After Treatment With Pathogen-Associated Molecular Pattern (Pamp) Elicitors, Xiao-Lin Chen, Xin Xie, Liye Wu, Caiyun Liu, Lirong Zeng, Xueping Zhou, Feng Luo, Guo-Liang Wang, Wende Liu

Department of Plant Pathology: Faculty Publications

Reversible protein ubiquitination plays essential roles in regulating cellular processes. Although many reports have described the functions of ubiquitination in plant defense responses, few have focused on global changes in the ubiquitome. To better understand the regulatory roles of ubiquitination in rice pattern-triggered immunity (PTI), we investigated the ubiquitome of rice seedlings after treatment with two pathogen-associated molecular patterns, the fungal-derived chitin or the bacterialderived flg22, using label-free quantitative proteomics. In chitin-treated samples, 144 and 167 lysine-ubiquitination sites in 121 and 162 proteins showed increased and decreased ubiquitination, respectively. In flg22-treated samples, 151 and 179 lysine-ubiquitination sites in 118 and …

Profile Of Genetically Modified Plants Authorized In Mexico, Mayra Teresa Garcia Ruiz, Aaron N. Knapp, Hernan Garcia Ruiz

Department of Plant Pathology: Faculty Publications

Mexico is a center of origin for several economically important plants including maize, cotton, and cocoa. Maize represents more than a food crop, has been declared a biological, cultural, agricultural and economic patrimony, and is linked to the national identity of Mexicans. In this review, we describe the historic and current use of genetically modified plants in Mexico and factors that contributed to the development of the biosafety regulation. We developed a database containing all permit applications received by the government to release genetically modified plants. A temporal and geographical analysis identified the plant species that have been authorized for …

Wheat Streak Mosaic Virus: A Century Old Virus With Rising Importance Worldwide, Khushwant Singh, Stephen N. Wegulo, Anna Skoracka, Jiban Kumar Kundu

Department of Plant Pathology: Faculty Publications

Wheat streak mosaic virus (WSMV) causes wheat streak mosaic, a disease of cereals and grasses that threatens wheat production worldwide. It is a monopartite, positive-sense, single-stranded RNA virus and the type member of the genus Tritimovirus in the family Potyviridae. The only known vector is the wheat curl mite (WCM, Aceria tosichella), recently identified as a species complex of biotypes differing in virus transmission. Low rates of seed transmission have been reported. Infected plants are stunted and have a yellow mosaic of parallel discontinuous streaks on the leaves. In the autumn, WCMs move from WSMV-infected volunteer wheat and other grass …

Determination Of Vegetative Development And Flower Effect Of Transgenic Nicotiana Tabacum, Hasan Pinar, Aydin Uyun, Semih Yilmaz, Nedim Mutlu, Amitava Mitra

Department of Plant Pathology: Faculty Publications

Species and varieties in the Nicotiana genus may respond differently to photoperiodism in terms of flowering time control. These are classified as short day, neutral day and long day plants. One of these, Nicotiana tabacum cv. The Xanthi nc is a genotype which can flowering on a long day. But the genotype Kanamycin resistance can open its flowers in January when the resistance gene is transferred to the plant. On the other hand, the effect of rootstock to scion in plant species is very important. One of the issues raised is whether the transgenic plant in the rootstock is effect …

Characterization Of Neofabraea Actinidiae And N. Brasiliensis As Causal Agents Of Apple Bull’S-Eye Rot In Southern Brazil, Amauri Bogo, Carla C. Comparin, Rosa M. Valdebenito Sanhueza, Patricia Ritschel, Ricardo T. Casa, Fábio N. Silva, Sydney E. Everhart

Department of Plant Pathology: Faculty Publications

The causal agents of apple bull’s-eye rot in southern Brazil have recently been described as Neofabraea actinidiae and N. brasiliensis. Isolates of both species were evaluated for response of mycelial growth index (MGI) to different temperatures, enzyme production, mycelial growth inhibition and effective concentrations (EC₅₀ and EC₁₀₀) of the fungicides triflumizole, pyrimethanil and thiophanate methyl, as well as aggressiveness on fruits of ‘Fuji’ hybrid and ‘Pink Lady’. There was significantly lower mycelium growth in N. brasiliensis compared with N. actinidiae at all temperatures tested. Neither species grew at 3 and 32°C. There were minor differences in …

Intermolecular Rna Recombination Occurs At Different Frequencies In Alternate Forms Of Brome Mosaic Virus Rna Replication Compartments, Hernan Garcia-Ruiz, Arturo Diaz, Paul Ahlquist

Department of Plant Pathology: Faculty Publications

Positive-strand RNA viruses replicate their genomes in membrane-bound replication compartments. Brome mosaic virus (BMV) replicates in vesicular invaginations of the endoplasmic reticulum membrane. BMV has served as a productive model system to study processes like virus-host interactions, RNA replication and recombination. Here we present multiple lines of evidence showing that the structure of the viral RNA replication compartments plays a fundamental role and that recruitment of parental RNAs to a common replication compartment is a limiting step in intermolecular RNA recombination. We show that a previously defined requirement for an RNA recruitment element on both parental RNAs is not to …

Clean Low-Biomass Procedures And Their Application To Ancient Ice Core Microorganisms, Zhi-Ping Zhong, Natalie E. Solonenko, Maria C. Gazitua, Donald V. Kenny, Ellen Mosley-Thompson, Virginia I. Rich, James L. Van Etten, Lonnie G. Thompson, Matthew B. Sullivan

Department of Plant Pathology: Faculty Publications

Microorganisms in glacier ice provide tens to hundreds of thousands of years archive for a changing climate and microbial responses to it. Analyzing ancient ice is impeded by technical issues, including limited ice, low biomass, and contamination. While many approaches have been evaluated and advanced to remove contaminants on ice core surfaces, few studies leverage modern sequencing to establish in silico decontamination protocols for glacier ice. Here we sought to apply such “clean” sampling techniques with in silico decontamination approaches used elsewhere to investigate microorganisms archived in ice at ~41 (D41, ~20,000 years) and ~49 m (D49, ~30,000 years) depth …

Tor-Autophagy Branch Signaling Via Imp1 Dictates Plant-Microbe Biotrophic Interface Longevity, Guangchao Sun, Christian Elowsky, Gang Li, Richard Wilson

Department of Plant Pathology: Faculty Publications

Like other intracellular eukaryotic phytopathogens, the devastating rice blast fungus Magnaporthe (Pyricularia) oryzae first infects living host cells by elaborating invasive hyphae (IH) surrounded by a plant-derived membrane. This forms an extended biotrophic interface enclosing an apoplastic compartment into which fungal effectors can be deployed to evade host detection. M. oryzae also forms a focal, plant membrane-rich structure, the biotrophic interfacial complex (BIC), that accumulates cytoplasmic effectors for translocation into host cells. Molecular decision-making processes integrating fungal growth and metabolism in host cells with interface function and dynamics are unknown. Here, we report unanticipated roles for the M. …

Field Evaluation Of Commercially Available Small Unmanned Aircraft Crop Spray Systems, Wayne Woldt, D. Martin, M. Latheef, Greg Kruger, Robert J. Wright, Justin Mcmechan, Christopher A. Proctor, Tamra Jackson-Ziems

Department of Plant Pathology: Faculty Publications

Agricultural research and development on small unmanned aircraft systems (UAS) has been directed toward UAS enabled sensing to detect features of interest. While compelling, there is an immediate need to increase the breadth and depth of UAS-based research, to move beyond sensing, and explore active intervention in agricultural production systems. This paper is focused on the concept of crop protection through ultra-precise, unmanned aerial application systems, and seeks to initiate research discussion in this important area of opportunity. Toward this end, two different, commercially available, small Unmanned Aerial Application Systems (sUAAS - defined as less than 55 lbs. maximum take-off …