In JoVE (1)

Other Publications (16)

Articles by William Underwood in JoVE

Other articles by William Underwood on PubMed

Genome-wide Transcriptional Analysis of the Arabidopsis Thaliana Interaction with the Plant Pathogen Pseudomonas Syringae Pv. Tomato DC3000 and the Human Pathogen Escherichia Coli O157:H7

The Plant Journal : for Cell and Molecular Biology. Apr, 2006  |  Pubmed ID: 16553894

Pseudomonas syringae pv. tomato DC3000 (Pst) is a virulent pathogen that causes disease on tomato and Arabidopsis. The type III secretion system (TTSS) plays a key role in pathogenesis by translocating virulence effectors from the bacteria into the plant host cell, while the phytotoxin coronatine (COR) contributes to virulence and disease symptom development. Recent studies suggest that both the TTSS and COR are involved in the suppression of host basal defenses. However, little is known about the interplay between the host gene expression changes associated with basal defenses and the virulence activities of the TTSS and COR during infection. In this study, we used the Affymetrix full genome chip to determine the Arabidopsis transcriptome associated with basal defense to Pst DC3000 hrp mutants and the human pathogenic bacterium Escherichia coli O157:H7. We then used Pst DC3000 virulence mutants to characterize Arabidopsis transcriptional responses to the action of hrp-regulated virulence factors (e.g. TTSS and COR) during bacterial infection. Additionally, we used bacterial fliC mutants to assess the role of the pathogen-associated molecular pattern flagellin in induction of basal defense-associated transcriptional responses. In total, our global gene expression analysis identified 2800 Arabidopsis genes that are reproducibly regulated in response to bacterial pathogen inoculation. Regulation of these genes provides a molecular signature for Arabidopsis basal defense to plant and human pathogenic bacteria, and illustrates both common and distinct global virulence effects of the TTSS, COR, and possibly other hrp-regulated virulence factors during Pst DC3000 infection.

Plant Stomata Function in Innate Immunity Against Bacterial Invasion

Cell. Sep, 2006  |  Pubmed ID: 16959575

Microbial entry into host tissue is a critical first step in causing infection in animals and plants. In plants, it has been assumed that microscopic surface openings, such as stomata, serve as passive ports of bacterial entry during infection. Surprisingly, we found that stomatal closure is part of a plant innate immune response to restrict bacterial invasion. Stomatal guard cells of Arabidopsis perceive bacterial surface molecules, which requires the FLS2 receptor, production of nitric oxide, and the guard-cell-specific OST1 kinase. To circumvent this innate immune response, plant pathogenic bacteria have evolved specific virulence factors to effectively cause stomatal reopening as an important pathogenesis strategy. We provide evidence that supports a model in which stomata, as part of an integral innate immune system, act as a barrier against bacterial infection.

Role of Plant Stomata in Bacterial Invasion

Cellular Microbiology. Jul, 2007  |  Pubmed ID: 17419713

Stomata are microscopic pores in the epidermis of the aerial parts of terrestrial plants. These pores are essential for photosynthesis, as they allow CO(2) to diffuse into the plant. The size of the stomatal pore changes in response to environmental conditions, such as light intensity, air humidity and CO(2) concentrations, as part of the plant's adaptation to maximize photosynthetic efficiency and, at the same time, to minimize water loss. Historically, stomata have been considered as passive portal of entry for plant pathogenic bacteria. However, recent studies suggest that stomata can play an active role in restricting bacterial invasion as part of the plant innate immune system. Some plant pathogens have evolved specific virulence factors to overcome stomata-based defence. Interestingly, many bacterial disease outbreaks require high humidity, rain, or frost damage, which could promote stomatal opening and/or bypass stomatal defence by creating wounds as alternative entry sites. Further studies on microbial and environmental regulation of stomata-based defence should fill gaps in our understanding of bacterial pathogenesis, disease epidemiology and phyllosphere microbiology.

The Pseudomonas Syringae Type III Effector Tyrosine Phosphatase HopAO1 Suppresses Innate Immunity in Arabidopsis Thaliana

The Plant Journal : for Cell and Molecular Biology. Nov, 2007  |  Pubmed ID: 17877704

The bacterial pathogen Pseudomonas syringae pv. tomato (Pst) strain DC3000 infects tomato and Arabidopsis plants, and is a model for studying the molecular basis of bacterial disease. Pst DC3000 secretes a battery of largely uncharacterized effector proteins into host cells via a type-III secretion system (TTSS). Little is currently known about the molecular mechanisms by which individual TTSS effectors promote virulence. The effector HopAO1 has similarity to protein tyrosine phosphatases, including a conserved catalytic site, and suppresses the hypersensitive response (HR) in some non-host plants. Whether HopAO1 has a similar effect in the host Arabidopsis is not clear. Here, we show that transgenic expression of HopAO1 in Arabidopsis suppresses callose deposition elicited by the Pst DC3000 hrpA mutant, and allows the normally non-pathogenic hrpA mutant to multiply within the leaf tissue. HopAO1 also suppresses resistance to Pst DC3000 induced by flg22, a pathogen-associated molecular pattern (PAMP). However, HopAO1 does not suppress the HR triggered by several classical avirulence genes. These results suggest that HopAO1 targets primarily PAMP-induced innate immunity in Arabidopsis. The virulence function of HopAO1 is dependent on an intact phosphatase catalytic site, as transgenic plants expressing a catalytically inactive derivative do not show these effects. Intriguingly, expression of the catalytically inactive HopAO1 has a dominant-negative effect on the function of the wild-type HopAO1. Analysis of mitogen-activated protein kinase (MAPK) activity suggests that HopAO1 targets a step downstream or independent of MAPK activation. Genome-wide expression analysis revealed that expression of several well-known defense genes was suppressed in hrpA mutant-infected HopAO1 transgenic plants.

Role of Stomata in Plant Innate Immunity and Foliar Bacterial Diseases

Annual Review of Phytopathology. 2008  |  Pubmed ID: 18422426

Pathogen entry into host tissue is a critical first step in causing infection. For foliar bacterial plant pathogens, natural surface openings, such as stomata, are important entry sites. Historically, these surface openings have been considered as passive portals of entry for plant pathogenic bacteria. However, recent studies have shown that stomata can play an active role in limiting bacterial invasion as part of the plant innate immune system. As a counter-defense, the plant pathogen Pseudomonas syringae pv. tomato DC3000 uses the virulence factor coronatine to actively open stomata. In nature, many foliar bacterial disease outbreaks require high humidity, rain, or storms, which could favor stomatal opening and/or bypass stomatal defense by creating wounds as alternative entry sites. Further studies on microbial and environmental regulation of stomatal closure and opening could fill gaps in our understanding of bacterial pathogenesis, disease epidemiology, and microbiology of the phyllosphere.

Focal Accumulation of Defences at Sites of Fungal Pathogen Attack

Journal of Experimental Botany. 2008  |  Pubmed ID: 18703493

Plants resist attack by haustorium-forming biotrophic and hemi-biotrophic fungi through fortification of the cell wall to prevent penetration through the wall and the subsequent establishment of haustorial feeding structures by the fungus. While the existence of cell wall-based defences has been known for many years, only recently have the molecular components contributing to such defences been identified. Forward genetic screens identified Arabidopsis mutants impaired in penetration resistance to powdery mildew fungi that were normally halted at the cell wall. Several loci contributing to penetration resistance have been identified and a common feature is the striking focal accumulation of proteins associated with penetration resistance at sites of interaction with fungal appressoria and penetration pegs. The focal accumulation of defence-related proteins and the deposition of cell wall reinforcements at sites of attempted fungal penetration represent an example of cell polarization and raise many questions of relevance, not only to plant pathology but also to general cell biology.

A House is Not a Home: Keeping Patients at the Center of Practice Redesign

Health Affairs (Project Hope). Sep-Oct, 2008  |  Pubmed ID: 18780904

The "patient-centered medical home" has been promoted as an enhanced model of primary care. Based on a literature review and interviews with practicing physicians, we find that medical home advocates and physicians have somewhat different, although not necessarily inconsistent, expectations of what the medical home should accomplish-from greater responsiveness to the needs of all patients to increased focus on care management for patients with chronic conditions. As the medical home concept is further developed, it will be important to not overemphasize redesign of practices at the expense of patient-centered care, which is the hallmark of excellent primary care.

Mitogen-activated Protein Kinases 3 and 6 Are Required for Full Priming of Stress Responses in Arabidopsis Thaliana

The Plant Cell. Mar, 2009  |  Pubmed ID: 19318610

In plants and animals, induced resistance (IR) to biotic and abiotic stress is associated with priming of cells for faster and stronger activation of defense responses. It has been hypothesized that cell priming involves accumulation of latent signaling components that are not used until challenge exposure to stress. However, the identity of such signaling components has remained elusive. Here, we show that during development of chemically induced resistance in Arabidopsis thaliana, priming is associated with accumulation of mRNA and inactive proteins of mitogen-activated protein kinases (MPKs), MPK3 and MPK6. Upon challenge exposure to biotic or abiotic stress, these two enzymes were more strongly activated in primed plants than in nonprimed plants. This elevated activation was linked to enhanced defense gene expression and development of IR. Strong elicitation of stress-induced MPK3 and MPK6 activity is also seen in the constitutive priming mutant edr1, while activity was attenuated in the priming-deficient npr1 mutant. Moreover, priming of defense gene expression and IR were lost or reduced in mpk3 or mpk6 mutants. Our findings argue that prestress deposition of the signaling components MPK3 and MPK6 is a critical step in priming plants for full induction of defense responses during IR.

Sugar Transporters for Intercellular Exchange and Nutrition of Pathogens

Nature. Nov, 2010  |  Pubmed ID: 21107422

Sugar efflux transporters are essential for the maintenance of animal blood glucose levels, plant nectar production, and plant seed and pollen development. Despite broad biological importance, the identity of sugar efflux transporters has remained elusive. Using optical glucose sensors, we identified a new class of sugar transporters, named SWEETs, and show that at least six out of seventeen Arabidopsis, two out of over twenty rice and two out of seven homologues in Caenorhabditis elegans, and the single copy human protein, mediate glucose transport. Arabidopsis SWEET8 is essential for pollen viability, and the rice homologues SWEET11 and SWEET14 are specifically exploited by bacterial pathogens for virulence by means of direct binding of a bacterial effector to the SWEET promoter. Bacterial symbionts and fungal and bacterial pathogens induce the expression of different SWEET genes, indicating that the sugar efflux function of SWEET transporters is probably targeted by pathogens and symbionts for nutritional gain. The metazoan homologues may be involved in sugar efflux from intestinal, liver, epididymis and mammary cells.

A Simple Intervention Promoting Patient Safety Improvements in Small Internal Medicine Practices

Quality in Primary Care. 2010  |  Pubmed ID: 21114911

small primary care practices may face difficulties in staying abreast of patient safety recommendations and implementing them. Some safety issues, however, may be easily and inexpensively addressed, given the necessary information on what is required.

Visualizing Cellular Dynamics in Plant-microbe Interactions Using Fluorescent-tagged Proteins

Methods in Molecular Biology (Clifton, N.J.). 2011  |  Pubmed ID: 21359815

Interactions between plant cells and microbial pathogens involve highly dynamic processes of cellular trafficking and reorganization. Substantial advances in imaging technologies, including the discovery and widespread use of fluorescent proteins as tags as well as advances in laser-based confocal microscopy have provided the first glimpses of the dynamic nature of the processes of defense and pathogenicity. Prior to the development of these techniques, high resolution imaging by electron microscopy gave only a static picture of these dynamic events and live cell imaging was significantly limited in resolution as well as the availability of relevant stains and markers. The incorporation of fluorescent protein fusions and laser-based confocal microscopy into studies of plant-microbe interactions has opened the door to fascinating new questions about the cellular response to attempted infection. Additionally, studies of cellular responses to pathogen infection may lead to new knowledge about fundamental processes in plant cells, such as mechanisms underlying subcellular trafficking and targeting of proteins and other molecules.

Design of a Quality and Performance Improvement Project for Small Primary Care Practices: Reflections on the Center for Practice Innovation

Quality in Primary Care. 2011  |  Pubmed ID: 21703112

Small practices often lack the human, financial and technical resources to make necessary practice improvements and infrastructure investments in order to achieve sustainable change that promotes quality and efficiency.

The Plant Cell Wall: a Dynamic Barrier Against Pathogen Invasion

Frontiers in Plant Science. 2012  |  Pubmed ID: 22639669

Prospective plant pathogens must overcome the physical barrier presented by the plant cell wall. In addition to being a preformed, passive barrier limiting access of pathogens to plant cells, the cell wall is actively remodeled and reinforced specifically at discrete sites of interaction with potentially pathogenic microbes. Active reinforcement of the cell wall through the deposition of cell wall appositions, referred to as papillae, is an early response to perception of numerous categories of pathogens including fungi and bacteria. Rapid deposition of papillae is generally correlated with resistance to fungal pathogens that attempt to penetrate plant cell walls for the establishment of feeding structures. Despite the ubiquity and apparent importance of this early defense response, relatively little is known about the underlying molecular mechanisms and cellular processes involved in the targeting and assembly of papillae. This review summarizes recent advances in our understanding of cell wall-associated defenses induced by pathogen perception as well as the impact of changes in cell wall polymers on interactions with pathogens and highlights significant unanswered questions driving future research in the area.

Induction and Suppression of PEN3 Focal Accumulation During Pseudomonas Syringae Pv. Tomato DC3000 Infection of Arabidopsis

Molecular Plant-microbe Interactions : MPMI. Aug, 2013  |  Pubmed ID: 23815470

The pleiotropic drug resistance (PDR) proteins belong to the super-family of ATP-binding cassette (ABC) transporters. AtPDR8, also called PEN3, is required for penetration resistance of Arabidopsis to nonadapted powdery mildew fungi. During fungal infection, plasma-membrane-localized PEN3 is concentrated at fungal entry sites, as part of the plant's focal immune response. Here, we show that the pen3 mutant is compromised in resistance to the bacterial pathogen Pseudomonas syringae pv. tomato DC3000. P. syringae pv. tomato DC3000 infection or treatment with a flagellin-derived peptide, flg22, induced strong focal accumulation of PEN3-green fluorescent protein. Interestingly, after an initial induction of PEN3 accumulation, P. syringae pv. tomato DC3000 but not the type-III-secretion-deficient mutant hrcC could suppress PEN3 accumulation. Moreover, transgenic overexpression of the P. syringae pv. tomato DC3000 effector AvrPto was sufficient to suppress PEN3 focal accumulation in response to flg22. Analyses of P. syringae pv. tomato DC3000 effector deletion mutants showed that individual effectors, including AvrPto, appear to be insufficient to suppress PEN3 accumulation when delivered by bacteria, suggesting a requirement for a combined action of multiple effectors. Collectively, our results indicate that PEN3 plays a positive role in plant resistance to a bacterial pathogen and show that focal accumulation of PEN3 protein may be a useful cellular response marker for the Arabidopsis-P. syringae interaction.

Perception of Conserved Pathogen Elicitors at the Plasma Membrane Leads to Relocalization of the Arabidopsis PEN3 Transporter

Proceedings of the National Academy of Sciences of the United States of America. Jul, 2013  |  Pubmed ID: 23836668

The Arabidopsis penetration resistance 3 (PEN3) ATP binding cassette transporter participates in nonhost resistance to fungal and oomycete pathogens and is required for full penetration resistance to the barley powdery mildew Blumeria graminis f. sp. hordei. PEN3 resides in the plasma membrane and is recruited to sites of attempted penetration by invading fungal appressoria, where the transporter shows strong focal accumulation. We report that recruitment of PEN3 to sites of pathogen detection is triggered by perception of pathogen-associated molecular patterns, such as flagellin and chitin. PEN3 recruitment requires the corresponding pattern recognition receptors but does not require the BAK1 coreceptor. Pathogen- and pathogen-associated molecular pattern-induced focal accumulation of PEN3 and the penetration resistance 1 (PEN1) syntaxin show differential sensitivity to specific pharmacological inhibitors, indicating distinct mechanisms for recruitment of these defense-associated proteins to the host-pathogen interface. Focal accumulation of PEN3 requires actin but is not affected by inhibitors of microtubule polymerization, secretory trafficking, or protein synthesis, and plasmolysis experiments indicate that accumulation of PEN3 occurs outside of the plasma membrane within papillae. Our results implicate pattern recognition receptors in the recruitment of defense-related proteins to sites of pathogen detection. Additionally, the process through which PEN3 is recruited to the host-pathogen interface is independent of new protein synthesis and BFA-sensitive secretory trafficking events, suggesting that existing PEN3 is redirected through an unknown trafficking pathway to sites of pathogen detection for export into papillae.

Contributions of Host Cellular Trafficking and Organization to the Outcomes of Plant-pathogen Interactions

Seminars in Cell & Developmental Biology. Aug, 2016  |  Pubmed ID: 27216829

In recent years it has become increasingly apparent that dynamic changes in protein localization, membrane trafficking pathways, and cellular organization play a major role in determining the outcome of interactions between plants and pathogenic microorganisms. Plants have evolved sophisticated perception systems to recognize the presence of potentially pathogenic microorganisms via the detection of non-self or modified-self elicitor molecules, pathogen virulence factors, or the activities of such virulence factors. Dynamic changes to host cellular organization and membrane trafficking pathways play pivotal roles in detection and signaling by plant immune receptors and are vital for the execution of spatially targeted defense responses to thwart invasion by potential pathogens. Conversely, from the perspective of the pathogen, the ability to manipulate plant cellular organization and trafficking processes to establish infection structures and deliver virulence factors is a major determinant of pathogen success. This review summarizes selected topics that illustrate how dynamic changes in host cellular trafficking and organization shape the outcomes of diverse plant-pathogen interactions and addresses ways in which our rapidly expanding knowledge of the cell biological processes that contribute to immunity or infection may influence efforts to improve plant disease resistance.

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