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Sample records for em xylella fastidiosa

  1. Morphological evidence for phages of Xylella fastidiosa

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Through transmission electron microscopy (TEM), phage particles of Xylella fastidiosa strain Temecula-1 grown in PW broth were observed in ultrathin sections of bacterial cell-containing low speed centrifugation pellets and in partially purified preparations from CsCl equilibrium centrifugation dens...

  2. Detection of small RNAs in Xylella fastidiosa

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Non-coding small RNAs (sRNAs) are regarded as ubiquitous regulatory elements in bacteria. For Xylella fastidiosa, a plant pathogen causing many economically important crop diseases, research attention to sRNAs has been limited. With the availability of whole genome sequences and increasing bioinfor...

  3. Characterization of novel secreted proteins from Xylella fastidiosa

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Xylella fastidiosa is a bacterium that causes disease of agriculturally important crops, including Pierce’s disease of grapevine. Little is known about virulence factors that are necessary for X. fastidiosa to grow and cause disease in the xylem vessels of a plant host. Any protein secreted by the b...

  4. Survey of phages in Xylella fastidiosa almond leaf scorch strains

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Xylella fastidiosa causes almond leaf scorch disease (ALSD) in California. The bacterium is nutritionally fastidious and difficult to culture in artificial media. Therefore, studies on the biology of X. fastidiosa are highly challenging. As a result, our knowledge about the pathogen and ALSD is limi...

  5. Identification of novel secreted virulence factors from Xylella fastidiosa

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Xylella fastidiosa is a bacterium that causes Pierce’s disease (PD) of grapevine and other leaf scorch diseases of agriculturally important crops. Little is known about virulence factors that are necessary for X. fastidiosa to grow and cause disease in the xylem vessels of a plant host. Any protein ...

  6. Genomic Characterization of a Phage in Xylella fastidiosa Almond Leaf Scorch Strain

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Xylella fastidiosa causes almond leaf scorch disease (ALSD) in California. The bacterium is nutritionally fastidious. Therefore, studies on the biology of X. fastidiosa are highly challenging. We previously reported the morphological observation of bacteriophages from X. fastidiosa culture. Current ...

  7. Xylella fastidiosa esterase rather than hydroxynitrile lyase.

    PubMed

    Torrelo, Guzman; Ribeiro de Souza, Fayene Zeferino; Carrilho, Emanuel; Hanefeld, Ulf

    2015-03-01

    In 2009, we reported that the product of the gene SCJ21.16 (XFa0032) from Xylella fastidiosa, a xylem-restricted plant pathogen that causes a range of diseases in several important crops, encodes a protein (XfHNL) with putative hydroxynitrile lyase activity. Sequence analysis and activity tests indicated that XfHNL exhibits an α/β-hydrolase fold and could be classified as a member of the family of FAD-independent HNLs. Here we provide a more detailed sequence analysis and new experimental data. Using pure heterologously expressed XfHNL we show that this enzyme cannot catalyse the cleavage/synthesis of mandelonitrile and that this protein is in fact a non-enantioselective esterase. Homology modelling and ligand docking simulations were used to study the active site and support these results. This finding could help elucidate the common ancestor of esterases and hydroxynitrile lyases with an α/β -hydrolase fold. PMID:25684099

  8. Draft genome sequence of Xylella fastidiosa subsp. fastidiosa strain Stag’s Leap

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Xylella fastidiosa subsp. fastidiosa causes Pierce’s disease of grapevine. Presented here is the draft genome sequence of the Stag’s Leap strain, previously used in pathogenicity/virulence assays to evaluate grapevine germplasm bearing Pierce’s disease....

  9. Draft Genome Sequence of Xylella fastidiosa subsp. fastidiosa Strain Stag’s Leap

    PubMed Central

    Wu, F.; Zheng, Z.; Deng, X.; Burbank, L. P.; Stenger, D. C.

    2016-01-01

    Xylella fastidiosa subsp. fastidiosa causes Pierce’s disease of grapevine. Presented here is the draft genome sequence of the Stag’s Leap strain, previously used in pathogenicity/virulence assays to evaluate grapevine germplasm bearing Pierce’s disease resistance and a phenotypic assessment of knockout mutants to determine gene function. PMID:27103713

  10. Fractal analysis of Xylella fastidiosa biofilm formation

    NASA Astrophysics Data System (ADS)

    Moreau, A. L. D.; Lorite, G. S.; Rodrigues, C. M.; Souza, A. A.; Cotta, M. A.

    2009-07-01

    We have investigated the growth process of Xylella fastidiosa biofilms inoculated on a glass. The size and the distance between biofilms were analyzed by optical images; a fractal analysis was carried out using scaling concepts and atomic force microscopy images. We observed that different biofilms show similar fractal characteristics, although morphological variations can be identified for different biofilm stages. Two types of structural patterns are suggested from the observed fractal dimensions Df. In the initial and final stages of biofilm formation, Df is 2.73±0.06 and 2.68±0.06, respectively, while in the maturation stage, Df=2.57±0.08. These values suggest that the biofilm growth can be understood as an Eden model in the former case, while diffusion-limited aggregation (DLA) seems to dominate the maturation stage. Changes in the correlation length parallel to the surface were also observed; these results were correlated with the biofilm matrix formation, which can hinder nutrient diffusion and thus create conditions to drive DLA growth.

  11. Development of a Model System to Elucidate Xylella Fastidiosa Pathogenesis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Pierce’s disease of grapes and almond leaf scorch are devastating diseases caused by the bacterium Xylella fastidiosa (Xf). To date, progress in determining the mechanisms of host plant susceptibility, tolerance or resistance has been slow, due in large part to the long generation time and limited a...

  12. Testing Xylella fastidiosa pathogenesis mutants in Arabidopsis thaliana

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The bacterium Xylella fastidiosa (Xf) causes Pierce's disease and a number of other plant diseases of significant economic impact. To date, progress determining mechanisms of host plant susceptibility, tolerance or resistance has been slow, due in large part to the long generation time and limited ...

  13. Searching for small RNA genes in Xylella fastidiosa genomes

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Bacterial non-coding small RNAs (sRNAs) have attracted considerable attention due to their ubiquitous nature and roles in controlling numerous cellular processes including survival, adaptation and pathogenesis. Xylella fastidiosa is a bacterial pathogen causing many economically important diseases s...

  14. Genomic characterization of a lysogenic phage from Xylella fastidiosa

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Xylella fastidiosa is an important pathogen causing disease of several economically important crops such as grape and almond in Central California. Despite intensive efforts to study this nutritionally fastidious pathogen, many biological traits of the bacterium such as bacteriophages remains poorl...

  15. Proteomic analysis of grapevines in response to Xylella fastidiosa infection

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Xylella fastidiosa (Xf) is the bacterial causal agent of Pierce’s disease (PD) of grapevines, as well as of other economically important diseases in a number of agronomic, horticultural and ornamental plants. The objective of this research was to tentatively identify proteins that are expressed in ...

  16. Extracellular Xylella fastidiosa genomic DNA enhances biofilm formation in vitro

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Xylella fastidiosa (Xf) is a Gram negative, xylem-limited bacterium that causes Pierce’s Disease (PD) of grapevine, as well as other diseases of economically important crops and landscape plants. Many bacteria produce large amounts of extracellular DNA, which may function as a matrix component in b...

  17. Functional identification of Xylella fastidiosa plasmid replication and stability factors

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Xylella fastidiosa (Xf) strain RIV11 harbors a 25 kbp plasmid (pXFRIV11) belonging to the incP1 incompatibility group. Replication and stability factors of pXFRIV11 were identified and used to construct plasmids able to replicate in both Xf and Escherichia coli. Sequences required for replication i...

  18. Selection of single chain variable fragments (scFv) against Xylella fastidiosa subsp. pauca by phage display

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Xylella fastidiosa is a gram-negative member of the gamma proteobacteria. Xylella fastidiosa subsp pauca causes citrus variegated chlorosis in Brazil and enjoys ‘select agent’ status in the United States. Antibody based detection assays are commercially available for Xylella fastidiosa, and are ef...

  19. Effects of grapevine sap phenolics on the in vitro growth of Xylella fastidiosa

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Pierce’s disease, caused by the bacterium Xylella fastidiosa, poses a serious threat to grape production in the United States. Previous work indicated that grapevines infected with Xylella fastidiosa respond by producing greater levels of phenolic compounds in xylem sap and tissues, presumably to l...

  20. Xylella fastidiosa Isolates from Both subsp. multiplex and fastidiosa Cause Disease on Southern Highbush Blueberry (Vaccinium sp.) Under Greenhouse Conditions.

    PubMed

    Oliver, J E; Cobine, P A; De La Fuente, L

    2015-07-01

    Xylella fastidiosa is a xylem-limited gram-negative plant pathogen that affects numerous crop species, including grape, citrus, peach, pecan, and almond. Recently, X. fastidiosa has also been found to be the cause of bacterial leaf scorch on blueberry in the southeastern United States. Thus far, all X. fastidiosa isolates obtained from infected blueberry have been classified as X. fastidiosa subsp. multiplex; however, X. fastidiosa subsp. fastidiosa isolates are also present in the southeastern United States and commonly cause Pierce's disease of grapevines. In this study, seven southeastern U.S. isolates of X. fastidiosa, including three X. fastidiosa subsp. fastidiosa isolates from grape, one X. fastidiosa subsp. fastidiosa isolate from elderberry, and three X. fastidiosa subsp. multiplex isolates from blueberry, were used to infect the southern highbush blueberry 'Rebel'. Following inoculation, all isolates colonized blueberry, and isolates from both X. fastidiosa subsp. multiplex and X. fastidiosa subsp. fastidiosa caused symptoms, including characteristic stem yellowing and leaf scorch symptoms as well as dieback of the stem tips. Two X. fastidiosa subsp. multiplex isolates from blueberry caused more severe symptoms than the other isolates examined, and infection with these two isolates also had a significant impact on host mineral nutrient content in sap and leaves. These findings have potential implications for understanding X. fastidiosa host adaptation and expansion and the development of emerging diseases caused by this bacterium. PMID:25738552

  1. MICROARRAY GENE EXPRESSION ANALYSIS OF GRAPE PLANTS IN RESPONSE TO XYLELLA FASTIDIOSA INFECTION

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Transcriptional profiling using a custom high-density microarray chip of 20,020 Vitis transcripts showed significant variations in responses between the susceptible and resistant genotypes to Xylella fastidiosa (Xf) infection. Differentially expressed transcripts reflecting spatial and temporal resp...

  2. Comparative Genomics- Identifying similarities and differences across three leafhopper vectors of Xylella fastidiosa

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Leafhoppers are the second most important vectors of agricultural diseases, thus we examined the gene expression across three leafhopper leafhoppers, Homalodisca vitripennis, Graphocephala atropunctata, and Oncometopia nigricans, which are vectors of the plant-infecting bacterium, Xylella fastidiosa...

  3. The Xylella fastidiosa PD1063 protein is secreted in association with outer membrane vesicles.

    PubMed

    Pierce, Brittany K; Voegel, Tanja; Kirkpatrick, Bruce C

    2014-01-01

    Xylella fastidiosa is a gram-negative, xylem-limited plant pathogenic bacterium that causes disease in a variety of economically important agricultural crops including Pierce's disease of grapevines. Xylella fastidiosa biofilms formed in the xylem vessels of plants play a key role in early colonization and pathogenicity by providing a protected niche and enhanced cell survival. Here we investigate the role of Xylella fastidiosa PD1063, the predicted ortholog of Xanthomonas oryzae pv. oryzae PXO_03968, which encodes an outer membrane protein. To assess the function of the Xylella fastidiosa ortholog, we created Xylella fastidiosa mutants deleted for PD1063 and then assessed biofilm formation, cell-cell aggregation and cell growth in vitro. We also assessed disease severity and pathogen titers in grapevines mechanically inoculated with the Xylella fastidiosa PD1063 mutant. We found a significant decrease in cell-cell aggregation among PD1063 mutants but no differences in cell growth, biofilm formation, disease severity or titers in planta. Based on the demonstration that Xanthomonas oryzae pv. oryzae PXO_03968 encodes an outer membrane protein, secreted in association with outer membrane vesicles, we predicted that PD1063 would also be secreted in a similar manner. Using anti-PD1063 antibodies, we found PD1063 in the supernatant and secreted in association with outer membrane vesicles. PD1063 purified from the supernatant, outer membrane fractions and outer membrane vesicles was 19.2 kD, corresponding to the predicted size of the processed protein. Our findings suggest Xylella fastidiosa PD1063 is not essential for development of Pierce's disease in Vitis vinifera grapevines although further research is required to determine the function of the PD1063 outer membrane protein in Xylella fastidiosa. PMID:25426629

  4. The Xylella fastidiosa PD1063 Protein Is Secreted in Association with Outer Membrane Vesicles

    PubMed Central

    Pierce, Brittany K.; Voegel, Tanja; Kirkpatrick, Bruce C.

    2014-01-01

    Xylella fastidiosa is a gram-negative, xylem-limited plant pathogenic bacterium that causes disease in a variety of economically important agricultural crops including Pierce's disease of grapevines. Xylella fastidiosa biofilms formed in the xylem vessels of plants play a key role in early colonization and pathogenicity by providing a protected niche and enhanced cell survival. Here we investigate the role of Xylella fastidiosa PD1063, the predicted ortholog of Xanthomonas oryzae pv. oryzae PXO_03968, which encodes an outer membrane protein. To assess the function of the Xylella fastidiosa ortholog, we created Xylella fastidiosa mutants deleted for PD1063 and then assessed biofilm formation, cell-cell aggregation and cell growth in vitro. We also assessed disease severity and pathogen titers in grapevines mechanically inoculated with the Xylella fastidiosa PD1063 mutant. We found a significant decrease in cell-cell aggregation among PD1063 mutants but no differences in cell growth, biofilm formation, disease severity or titers in planta. Based on the demonstration that Xanthomonas oryzae pv. oryzae PXO_03968 encodes an outer membrane protein, secreted in association with outer membrane vesicles, we predicted that PD1063 would also be secreted in a similar manner. Using anti-PD1063 antibodies, we found PD1063 in the supernatant and secreted in association with outer membrane vesicles. PD1063 purified from the supernatant, outer membrane fractions and outer membrane vesicles was 19.2 kD, corresponding to the predicted size of the processed protein. Our findings suggest Xylella fastidiosa PD1063 is not essential for development of Pierce's disease in Vitis vinifera grapevines although further research is required to determine the function of the PD1063 outer membrane protein in Xylella fastidiosa. PMID:25426629

  5. Xylella fastidiosa gene expression analysis by DNA microarrays

    PubMed Central

    2009-01-01

    Xylella fastidiosa genome sequencing has generated valuable data by identifying genes acting either on metabolic pathways or in associated pathogenicity and virulence. Based on available information on these genes, new strategies for studying their expression patterns, such as microarray technology, were employed. A total of 2,600 primer pairs were synthesized and then used to generate fragments using the PCR technique. The arrays were hybridized against cDNAs labeled during reverse transcription reactions and which were obtained from bacteria grown under two different conditions (liquid XDM2 and liquid BCYE). All data were statistically analyzed to verify which genes were differentially expressed. In addition to exploring conditions for X. fastidiosa genome-wide transcriptome analysis, the present work observed the differential expression of several classes of genes (energy, protein, amino acid and nucleotide metabolism, transport, degradation of substances, toxins and hypothetical proteins, among others). The understanding of expressed genes in these two different media will be useful in comprehending the metabolic characteristics of X. fastidiosa, and in evaluating how important certain genes are for the functioning and survival of these bacteria in plants. PMID:21637690

  6. Variations of whole genome sequences of Xylella fastidiosa strains within the same pathotype

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Xylella fastidiosa is a Gram negative and nutritionally fastidious plant pathogenic bacterium that causes almond leaf scorch disease (ALSD) and Pierce's disease (PD) of grapevine. X. fastidiosa strains from almond can be divided into two pathotypes: ALSD-PD, represented by strain M23, and ALSD-only,...

  7. Identification of novel secreted virulence factors from Xylella fastidiosa using a TRV expression system

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Xylella fastidiosa is a bacterium that causes leaf scorch diseases of agriculturally important crops including grapevines and almonds. Little is known about virulence factors that are necessary for X. fastidiosa to grow and cause disease in the xylem vessels of a plant host. Any protein secreted by ...

  8. Origins of the Xylella fastidiosa prophage-like regions and their impact in genome differentiation

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Xylella fastidiosa is a Gram negative plant pathogen causing many economically important diseases, and analyses of completely sequenced X. fastidiosa genome strains allowed the identification of many prophage-like elements and possibly phage remnants, accounting for up to 15% of the genome compositi...

  9. Validation of Small RNAs In Xylella fastidiosa by qRT-PCR

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Xylella fastidiosa causes many economically important crop diseases including almond leaf scorch disease and Pierce’ disease of grapevine. Although non-coding small RNAs (sRNAs) are regarded as ubiquitous regulatory elements in bacteria, research attention to sRNAs in X. fastidiosa has been limited...

  10. Evaluation of the Genetic Structure of Xylella fastidiosa Populations Collected from Almond orchards in California

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Xylella fastidiosa causes many economically important crop diseases including Pierce’s disease of grape and almond leaf scorch disease. A more detailed understanding of the genetic structure of X. fastidiosa populations is likely to aid development of novel management strategies. Current evidence i...

  11. Functional characterization of the role of the PilG in Xylella fastidiosa

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Type IV pili of Xylella fastidiosa are regulated by pilG, a chemotaxis regulator in the Pil-Chp operon involving signal transduction pathways. To elucidate the role of pilG in twitching motility and pathogenicity of X. fastidiosa, phenotypes of wild type, a pilG-mutant, and a complementary strain we...

  12. Evaluation of olive as a host of Xylella fastidiosa and associated sharpshooter vectors

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Olive (Olea europaea L.) trees exhibiting leaf scorch and/or branch dieback symptoms in California were surveyed for the xylem-limited, fastidious bacterium Xylella fastidiosa. Only ~17% of diseased trees tested positive for X. fastidiosa by PCR, and disease symptoms could not be attributed to X. fa...

  13. Characterization of Xylella fastidiosa pear leaf scorch strain in Taiwan through whole genome sequence analyses

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Xylella fastidiosa is a Gram-negative bacterium causing diseases in many economically important crops mostly in the Americas but also in Asia and Europe. A strain of X. fastidiosa was found to cause pear leaf scorch (PLS) disease in Taiwan in 1992. Because of nutritional fastidiousness, characteriza...

  14. Pathogen Special: Vibrio Cholerae, Pseudomonas Aeruginosa and Xylella Fastidiosa

    PubMed Central

    2000-01-01

    One could almost say that it is the latest fashion to sequence a bacterial genome. However, this would belittle the efforts of those working on these important organisms, whose data will greatly help those working on the prevention of disease in the fields of medicine and agriculture. In this feature we present a guided tour of the latest additions to the ‘sequenced microbes’ club. Vibrio cholerae is the causative agent of cholera, which is still a threat in countries with poor sanitation and unsafe drinking water. Pseudomonas aeruginosa is responsible for a large proportion of opportunistic human infections, typically infecting those with compromised immune systems, particularly cystic fibrosis patients, those patients on respirators and burn victims. Xylella fastidiosa is a plant pathogen that attacks citrus fruits by blocking the xylem, resulting in juiceless fruits of no commercial value. PMID:11119308

  15. Development of single chain variable fragment (scFv) antibodies against Xylella fastidiosa subsp. pauca by phage display

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Xylella fastidiosa is a member of the gamma proteobacteria. It is fastidious, insect-vectored and xylem-limited and causes a variety of diseases, some severe, on a wide range of economically important perennial crops, including grape and citrus. Xylella fastidiosa subsp pauca causes citrus variegat...

  16. THE ENDOPHYTE CURTOBACTERIUM FLACCUMFACIENS REDUCES SYMPTOMS CAUSED BY XYLELLA FASTIDIOSA IN CATHARANTHUS ROSEUSAN ENDOPHYTIC BACTERIUM FROM CITRUS

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Citrus variegated chlorosis (CVC) is a disease of sweet orange (Citrus sinensis (L.)) caused by Xylella fastidiosa subsp. pauca, a phytopathogenic bacterium that can infect all sweet orange cultivars. Sweet orange trees are sometimes observed to be infected by Xylella fastidiosa without showing seve...

  17. Diversity Evaluation of Xylella fastidiosa from Infected Olive Trees in Apulia (Southern Italy)

    PubMed Central

    Mang, Stefania M.; Frisullo, Salvatore; Elshafie, Hazem S.; Camele, Ippolito

    2016-01-01

    Olive culture is very important in the Mediterranean Basin. A severe outbreak of Olive Quick Decline Syndrome (OQDS) caused by Xylella fastidiosa infection was first noticed in 2013 on olive trees in the southern part of Apulia region (Lecce province, southern Italy). Studies were carried out for detection and diversity evaluation of the Apulian strain of Xylella fastidiosa. The presence of the pathogen in olive samples was detected by PCR amplifying the 16S rDNA, gyrase B subunit (gyrB) and HL hypothetical protein genes and single nucleotide polymorphisms (SNPs) assessment was performed to genotype X. fastidiosa. Twelve SNPs were recorded over gyrB and six SNPs were found for HL gene. Less variations were detected on 16S rDNA gene. Only gyrB and HL provided sufficient information for dividing the Apulian X. fastidiosa olive strains into subspecies. Using HL nucleotide sequences was possible to separate X. fastidiosa into subspecies pauca and fastidiosa. Whereas, nucleotide variation present on gyrB gene allowed separation of X. fastidiosa subsp. pauca from the other subspecies multiplex and fastidiosa. The X. fastidiosa strain from Apulia region was included into the subspecies pauca based on three genes phylogenetic analyses. PMID:27147930

  18. Diversity Evaluation of Xylella fastidiosa from Infected Olive Trees in Apulia (Southern Italy).

    PubMed

    Mang, Stefania M; Frisullo, Salvatore; Elshafie, Hazem S; Camele, Ippolito

    2016-04-01

    Olive culture is very important in the Mediterranean Basin. A severe outbreak of Olive Quick Decline Syndrome (OQDS) caused by Xylella fastidiosa infection was first noticed in 2013 on olive trees in the southern part of Apulia region (Lecce province, southern Italy). Studies were carried out for detection and diversity evaluation of the Apulian strain of Xylella fastidiosa. The presence of the pathogen in olive samples was detected by PCR amplifying the 16S rDNA, gyrase B subunit (gyrB) and HL hypothetical protein genes and single nucleotide polymorphisms (SNPs) assessment was performed to genotype X. fastidiosa. Twelve SNPs were recorded over gyrB and six SNPs were found for HL gene. Less variations were detected on 16S rDNA gene. Only gyrB and HL provided sufficient information for dividing the Apulian X. fastidiosa olive strains into subspecies. Using HL nucleotide sequences was possible to separate X. fastidiosa into subspecies pauca and fastidiosa. Whereas, nucleotide variation present on gyrB gene allowed separation of X. fastidiosa subsp. pauca from the other subspecies multiplex and fastidiosa. The X. fastidiosa strain from Apulia region was included into the subspecies pauca based on three genes phylogenetic analyses. PMID:27147930

  19. Survey of the auchenorrhyncha in the Salento Peninsula in search of putative vectors of Xylella fastidiosa Pauca Codiro strain

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Xylella fastidiosa was isolated in September 2013 from olive trees symptomatic for Olive Quick Decline Syndrome (OQDS) in Salento (Italy) and was identified as X. fastidiosa pauca CoDiRO strain. X. fastidiosa is comprised of a group of genetically diverse bacteria in the class Gammaproteobacteria th...

  20. Hierarchical Analysis and Diversity Studies of Xylella fastidiosa Populations in California by Multi-locus Simple Sequence Repeat Markers

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Xylella fastidiosa is the causative agent of Pierce’s Disease (PD) in grapevine. Using 18 simple sequence repeat (SSR) markers, we assessed variation within and between populations of X. fastidiosa isolated from grapevine in California. Eighty-three X. fastidiosa isolates from 15 populations presen...

  1. Structural and Biochemical Characterization of Peroxiredoxin Qβ from Xylella fastidiosa

    PubMed Central

    Horta, Bruno Brasil; de Oliveira, Marcos Antonio; Discola, Karen Fulan; Cussiol, José Renato Rosa; Netto, Luis Eduardo Soares

    2010-01-01

    The phytopathogenic bacterium Xylella fastidiosa is the etiological agent of various plant diseases. To survive under oxidative stress imposed by the host, microorganisms express antioxidant proteins, including cysteine-based peroxidases named peroxiredoxins. This work is a comprehensive analysis of the catalysis performed by PrxQ from X. fastidiosa (XfPrxQ) that belongs to a peroxiredoxin class still poorly characterized and previously considered as moderately reactive toward hydroperoxides. Contrary to these assumptions, our competitive kinetics studies have shown that the second-order rate constants of the peroxidase reactions of XfPrxQ with hydrogen peroxide and peroxynitrite are in the order of 107 and 106 m−1 s−1, respectively, which are as fast as the most efficient peroxidases. The XfPrxQ disulfides were only slightly reducible by dithiothreitol; therefore, the identification of a thioredoxin system as the probable biological reductant of XfPrxQ was a relevant finding. We also showed by site-specific mutagenesis and mass spectrometry that an intramolecular disulfide bond between Cys-47 and Cys-83 is generated during the catalytic cycle. Furthermore, we elucidated the crystal structure of XfPrxQ C47S in which Ser-47 and Cys-83 lie ∼12.3 Å apart. Therefore, significant conformational changes are required for disulfide bond formation. In fact, circular dichroism data indicated that there was a significant redox-dependent unfolding of α-helices, which is probably triggered by the peroxidatic cysteine oxidation. Finally, we proposed a model that takes data from this work as well data as from the literature into account. PMID:20335172

  2. Genome Sequence of a Xylella fastidiosa Strain Causing Sycamore Leaf Scorch Disease in Virginia.

    PubMed

    Guan, Wei; Shao, Jonathan; Davis, Robert E; Zhao, Tingchang; Huang, Qi

    2014-01-01

    Xylella fastidiosa causes bacterial leaf scorch in landscape trees including sycamore. We determined the draft genome of X. fastidiosa strain Sy-Va, isolated in Virginia from a sycamore tree displaying leaf scorch symptoms. The Sy-VA genome contains 2,477,829 bp, and has a G+C content of 51.64 mol%. PMID:25146135

  3. Multilocus sequence typing of Xylella fastidiosa causing Pierce's disease and oleander leaf scorch in the United States.

    PubMed

    Yuan, Xiaoli; Morano, Lisa; Bromley, Robin; Spring-Pearson, Senanu; Stouthamer, Richard; Nunney, Leonard

    2010-06-01

    Using a modified multilocus sequence typing (MLST) scheme for the bacterial plant pathogen Xylella fastidiosa based on the same seven housekeeping genes employed in a previously published MLST, we studied the genetic diversity of two subspecies, X. fastidiosa subsp. fastidiosa and X. fastidiosa subsp. sandyi, which cause Pierce's disease and oleander leaf scorch, respectively. Typing of 85 U.S. isolates (plus one from northern Mexico) of X. fastidiosa subsp. fastidiosa from 15 different plant hosts and 21 isolates of X. fastidiosa subsp. sandyi from 4 different hosts in California and Texas supported their subspecific status. Analysis using the MLST genes plus one cell-surface gene showed no significant genetic differentiation based on geography or host plant within either subspecies. Two cases of homologous recombination (with X. fastidiosa subsp. multiplex, the third U.S. subspecies) were detected in X. fastidiosa subsp. fastidiosa. Excluding recombination, MLST site polymorphism in X. fastidiosa subsp. fastidiosa (0.048%) and X. fastidiosa subsp. sandyi (0.000%) was substantially lower than in X. fastidiosa subsp. multiplex (0.240%), consistent with the hypothesis that X. fastidiosa subspp. fastidiosa and sandyi were introduced into the United States (probably just prior to 1880 and 1980, respectively). Using whole-genome analysis, we showed that MLST is more effective at genetic discrimination at the specific and subspecific level than other typing methods applied to X. fastidiosa. Moreover, MLST is the only technique effective in detecting recombination. PMID:20465416

  4. Draft Genome Sequence of the Xylella fastidiosa CoDiRO Strain.

    PubMed

    Giampetruzzi, Annalisa; Chiumenti, Michela; Saponari, Maria; Donvito, Giacinto; Italiano, Alessandro; Loconsole, Giuliana; Boscia, Donato; Cariddi, Corrado; Martelli, Giovanni Paolo; Saldarelli, Pasquale

    2015-01-01

    We determined the draft genome sequence of the Xylella fastidiosa CoDiRO strain, which has been isolated from olive plants in southern Italy (Apulia). It is associated with olive quick decline syndrome (OQDS) and characterized by extensive scorching and desiccation of leaves and twigs. PMID:25676759

  5. Leafhopper comparative genomics - Identifying similarities and differences across Leafhopper vectors of Xylella fastidiosa

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Leafhoppers, (Hemiptera) are considered the second most important group of plant pathogen vectors. Three leafhopper species which transmit Xylella fastidiosa, a plant pathogen of fruit crops, were examined by comparison of the available expressed sequence tags, ~43,400 ESTs (Hunter datasets, Nationa...

  6. High throughput PCR detection of Xylella fastidiosa directly from almond tissues

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Xylella fastidiosa, the causal agent of almond leaf scorch disease (ASLD), is currently re-emerging as a serious disease in California. Pathogen detection is a critical step for ALSD diagnosis because the disease symptoms are often confused with salt toxicity and other environmental stresses. We hav...

  7. Susceptibility of Prunus rootstock seedlings to Xylella fastidiosa strains isolated from almond in California

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Almond leaf scorch disease (ALSD) has a historical presence in California. ALSD is caused by the infection of Xylella fastidiosa, a nutritionally fastidious bacterium. In the past few years, the disease has re-emerged in the Central Valley of California. Like Pierce’s disease (PD) in grapes, ther...

  8. Proteomic analysis of grapevine tissues in response to Xylella fastidiosa infection

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Xylella fastidiosa (Xf) is the bacterial causal agent of Pierce’s disease (PD) as well as other economically important diseases in a number of agronomic, horticultural and ornamental plants. The objective of this research was to identify candidate proteins that are differentially expressed in grape...

  9. A conjugative 38kB plasmid is present in multiple subspecies of Xylella fastidiosa

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A ~38kB plasmid was present in the Riv5 strain of Xylella fastidiosa subsp. multiplex isolated from ornamental plum in southern California. This plasmid, pXF-RIV5, encodes a complete type IV secretion system necessary for conjugation and DNA transfer. pXF-RIV5 is almost identical to pXFAS01 from X. ...

  10. Habitat Effects on Population Density and Movement of Insect Vectors of Xylella fastidiosa in California, USA

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Xylella fastidiosa is a xylem-limited bacterium that causes disease in grapevines, almonds, citrus, pear, alfalfa, and many other economically important plants. In California, USA, the bacteria are transmitted by several species of leafhoppers including the cicadellids Draeculacephala minerva Ball a...

  11. Evaluation of Arabidopsis thaliana as a model host for Xylella fastidiosa

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Pierce’s disease of grapes and almond leaf scorch are diseases caused by the bacterium Xylella fastidiosa. To date, progress determining mechanisms of host plant susceptibility, tolerance or resistance has been slow, due in large part to the long generation time and limited available genetic resourc...

  12. Evaluation of Arabidopsis thaliana as a model host for Xylella fastidiosa

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The bacterium Xylella fastidiosa (Xf) causes a number of devastating plant diseases of significant economic impact. To date, progress determining mechanisms of host plant susceptibility, tolerance or resistance has been slow, due in large part to the long generation time and limited available genet...

  13. Draft genome sequence of Xylella fastidiosa supsp. multiplex strain Griffin-1 from Quercus rubra in Georgia

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The draft genome sequence of Xylella fastidiosa subsp. multiplex Strain Griffin-1 isolated from a red oak tree (Quercus rubra) in Georgia, U.S.A. is reported. The bacterium has a genome size of 2,387,314 bp with 51.7% G+C content and comprises 2,903 predicted open reading frames (ORFs), and 50 RNA g...

  14. Seasonal abundance of Draeculacephala minerva and other Xylella fastidiosa vectors in California almond orchards and vineyards

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Almond leaf scorch (ALS) is caused by the bacterium Xylella fastidiosa which is transmitted by xylem-feeding sharpshooters and spittlebugs. Reports of increased incidence of ALS-affected almonds in California prompted surveys in three almond-growing regions in California, from June 2003 to September...

  15. Whole genome sequence analyses of Xylella fastidiosa PD strains from different geographical regions

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Genome sequences were determined for two Pierce’s disease (PD) causing Xylella fastidiosa (Xf) strains, one from Florida and one from Taiwan. The Florida strain was ATCC 35879, the type of strain used as a standard reference for related taxonomy research. By contrast, the Taiwan strain used was only...

  16. Evidence supporting the Egestion-Salivation Hypothesis for inoculation of Xylella fastidiosa by sharpshooter vectors

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Despite more than 70 years of study, the mechanism of inoculation of semipersistent, foregut-borne plant pathogens by their vectors is still unknown. The best model system for these studies is inoculation of the Pierce’s disease bacterium, Xylella fastidiosa (Xf), by vectors such as the glassy-wing...

  17. Environmental factors affecting twitching motility, biofilm development, and aggregation by Xylella fastidiosa

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The bacterial pathogen Xylella fastidiosa causes many important plant diseases in different crops such as citrus, grapes, almond and coffee. While disease symptoms expressed by this pathogen are not completely understood, it is widely accepted that blockage of xylem vessels by aggregates of the bact...

  18. Specific detection and identification of Xylella fastidiosa strains causing oleander leaf scorch by polymerase chain reaction

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A pair of PCR primers, QH-OLS05/QH-OLS08, was developed that is specific for strains of Xylella fastidiosa causing oleander leaf scorch. The primers were designed based on DNA sequence of a randomly amplified polymorphic DNA (RAPD)-PCR product unique to oleander strains. The PCR assay using primer p...

  19. Identification and characterization of proteomic expression of grapevines in response to Xylella fastidiosa infection.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Xylella fastidiosa (Xf) is the bacterial causal agent of Pierce’s disease (PD) of grapevines, as well as of other economically important diseases in a number of agronomic, horticultural and ornamental plants. In this study, comparative proteomic analyses were carried out to identify proteins differ...

  20. Genomic insights into Xylella fastidiosa interactions with plant and insect hosts

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Utilization of genomic data and widespread availability of genomics tools are still incipient in plant pathology. The first genome of a plant pathogen, the bacterium Xylella fastidiosa, was only completed in 2000. Since then, a large number of bacterial plant pathogens have been sequenced, but much ...

  1. Evidence that explains absence of a latent period for Xylella fastidiosa in its sharpshooter vectors

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The glassy-winged sharpshooter (GWSS), Homalodisca vitripennis (Germar), and other sharpshooter (Cicadelline) leafhoppers transmit Xylella fastidiosa (Xf), the causative agent of Pierce’s disease of grapevine and other scorch diseases. Past research has supported that vectors have virtually no late...

  2. Characterization of Xylella fastidiosa popP gene required for pathogenicity

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Xylella fastidiosa (Xf) possesses a two component regulatory system (TCS) popP-popQ which differentially regulates genes in response to environmental stimuli. To elucidate the role of popP in Pierce’s disease (PD) of grapes, a site-directed deletion method and chromosome-based genetic complementatio...

  3. Seasonal Population Dynamics of Draeculacephala minerva (Hemiptera: Cicadellidae) and Transmission Efficiency of Xylella fastidiosa

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The green sharpshooter, Draeculacephala minerva Ball, is a very common and often abundant grass-feeding leafhopper in California. Its population dynamics and ability to transmit Xylella fastidiosa were monitored over a 2-year period in California’s San Joaquin Valley. Collections of individuals fr...

  4. A stable shuttle vector for Xylella fastidiosa based on an endogenous incP-1 plasmid

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Xylella fastidiosa (Xf) strain RIV11 harbors a 25 kbp plasmid (pXFRIV11) belonging to the incP1 incompatibility group. Replication and stability factors of pXFRIV11 were identified and used to construct plasmids able to propagate in both Xf and Escherichia coli. Sequences required for replication i...

  5. Identification of Proteomic Expression of Grapevines in Response to Xylella fastidiosa

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Xylella fastidiosa (Xf) is the xylem-limited plant pathogen that causes grape Pierce’s disease (PD) as well as other economically important diseases in a number of fruit and ornamental plants. The objective of this research is to identify differentially expressed proteins in grapevines that are inv...

  6. The DinJ/RelE toxin-antitoxin system suppresses virulence in Xylella fastidiosa

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Xylella fastidiosa, the causal agent of a number agriculturally important plant diseases, encodes multiple toxin-antitoxin (TA) systems. TA modules consist of a toxin protein co-expressed with a specific antitoxin, and are often acquired through horizontal gene transfer. Antitoxin molecules (RNA or ...

  7. The role of Xylella fastidiosa cold shock proteins in Pierce’s disease of grapes

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Pierce’s disease of grapevine, caused by the bacterial pathogen Xylella fastidiosa (Xf) is limited to warmer climates, and plant infection can be eliminated by cold winter conditions. Milder winters can increase the likelihood of pathogen persistence from one growing season to the next. Cold adaptat...

  8. Stable plasmid vectors for complementation of Xylella fastidiosa mutants in planta

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Current understanding of the mechanisms of Pierce’s Disease development has been significantly advanced by molecular genetic studies of the causal agent, Xylella fastidiosa (Xf). Plasmid vectors are an essential tool for studies of bacterial genetics and pathogenesis. However, most commonly used pla...

  9. VARIATION OF PSPB (PD1208) LOCUS AMONG CLOSELY RELATED XYLELLA FASTIDIOSA STRAINS

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Xylella fastidiosa is a Gram negative plant pathogenic bacterium causing grape Pierce’s disease (PD) and almond leaf scorch disease (ALSD). Two distinct genotypes are associated with ALSD. A-genotype strains cause ALSD but not PD. G-genotype strains cause both ALSD and PD. However, little is known a...

  10. Evaluation of Arabidopsis thaliana as an experimental host for Xylella fastidiosa

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Pierce’s disease of grapes and almond leaf scorch are diseases caused by the bacterium Xylella fastidiosa. To date, progress determining mechanisms of host plant susceptibility, tolerance or resistance has been slow, due in large part to the long generation time and limited available genetic resourc...

  11. Draft genome sequence of Xylella fastidiosa pear leaf scorch strain in Taiwan

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The draft genome sequence of Xylella fastidiosa pear leaf scorch strain (PLS229) isolated from pear cultivar Hengshan (Pyrus pyrifolia) in Taiwan is reported. The bacterium has a genome size of 2,733,013 bp with a G+C content of 53.1%. The PLS229 strain genome was annotated to have 3,259 open readin...

  12. Evaluation of Arabidopsis thaliana as a model host for Xylella fastidiosa

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Pierce’s disease of grapes and almond leaf scorch are devastating diseases caused by the bacterium Xylella fastidiosa (Xf). To date, progress in determining the mechanisms of host plant succeptibility, tolerance or resistance has been slow, due in large part to the long generation time and limited a...

  13. Induction of phenolic compounds in response to Xylella fastidiosa infection in five different grapevine cultivars

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Previously, Thompson seedless grapevines infected with Xylella fastidiosa (Xf), the causal agent of Pierce’s disease (PD), were observed to possess greater phenolic levels in xylem sap and tissues than non-infected plants shortly after inoculation. Plants often produce greater levels of phenolic com...

  14. Xylella fastidiosa Extracellular Genomic DNA May Play a Role For Enhancing Biofilm Formation In Vitro

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Xylella fastidiosa (Xf) produces extracellular DNA in PD3 liquid medium. This extracellular DNA may play a role in enhancing biofilm formation, a factor that is required by Xf to establish infection in host plants. Amounts of extracellular DNA generated by Xf in vitro were positively correlated with...

  15. Lack of evidence for transmission of Xylella fastidiosa from infected sweet orange seed

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Citrus variegated chlorosis is among the principle diseases that affect sweet orange in Brazil and Argentina, and is viewed as an emerging threat by the U.S. sweet orange industry. The disease is caused by the fastidious bacterium Xylella fastidiosa, and can be transmitted by both leafhopper insect...

  16. Genome Sequence of a Xylella fastidiosa Strain Causing Mulberry Leaf Scorch Disease in Maryland.

    PubMed

    Guan, Wei; Shao, Jonathan; Zhao, Tingchang; Huang, Qi

    2014-01-01

    Xylella fastidiosa causes bacterial leaf scorch in landscape trees, including mulberry. We determined the draft genome of the mulberry strain Mul-MD in order to gain a better understanding of the molecular basis of strain divergence, host specificity, nutrient requirements, and pathogenicity, as well as to develop genome-based specific detection methods. PMID:24604658

  17. Genomic Sequencing of Two Coffee-Infecting Strains of Xylella fastidiosa Isolated from Brazil.

    PubMed

    Alencar, Valquíria C; Barbosa, Deibs; Santos, Daiene S; Oliveira, Ana Cláudia F; de Oliveira, Regina C; Nunes, Luiz R

    2014-01-01

    Here, we describe the draft genome sequences of two Xylella fastidiosa strains: Xf6c and Xf32, which have been obtained from infected coffee plants in Brazil, and are associated with the disease known as coffee leaf scorch (CLS). PMID:24435874

  18. Draft Genome Sequence of CO33, a Coffee-Infecting Isolate of Xylella fastidiosa

    PubMed Central

    Loconsole, Giuliana; Boscia, Donato; Calzolari, Alessandra; Chiumenti, Michela; Martelli, Giovanni P.; Saldarelli, Pasquale; Almeida, Rodrigo P. P.; Saponari, Maria

    2015-01-01

    The draft genome sequence of Xylella fastidiosa CO33 isolate, retrieved from symptomatic leaves of coffee plant intercepted in northern Italy, is reported. The CO33 genome size is 2,681,926 bp with a GC content of 51.7%. PMID:26679584

  19. Glassy-winged sharpshooter can use a mechanical mechanism to inoculate Xylella fastidiosa into grapevines

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Xylem-feeding leafhoppers such as the glassy-winged sharpshooter, Homalodisca vitripennis (Cicadellidae: Cicadellinae), are thought to inoculate the bacterium Xylella fastidiosa (Xf) from colonies bound to cuticle of the sharpshooter’s functional foregut (precibarium and cibarium). The mechanism of ...

  20. Grapevine phenolics in xylem sap and tissues are significantly altered during infection by Xylella fastidiosa

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Pierce’s disease of grapevine (PD), caused by the bacterial pathogen Xylella fastidiosa (X.f.), remains a serious problem for grape production in California and elsewhere. This research examined induction of phenolic compounds in grapevines (cv. Thompson Seedless) infected with X.f. over a six month...

  1. Phenology of Xylella fastidiosa and its Vector, Draeculacephala minerva, in vegetation surrounding California Almond Nurseries

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Management of almond leaf scorch disease requires knowledge of all possible infection pathways. The disease is caused by the xylem-limited bacterium Xylella fastidiosa, which is transmitted by several species of leafhoppers. The objectives of this research were to elucidate the fate of bacteria in p...

  2. Xylella fastidiosa infection of grapevines affects xylem levels of phenolic compounds and pathogenesis-related proteins

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Pierce’s disease (PD), caused by the xylem-dwelling pathogen Xylella fastidiosa (X.f.), is a serious threat to grape production. The effects of X.f. infection six months post-inoculation on defense-associated proteins and phenolic compounds found in xylem sap and tissue were evaluated. Defense-assoc...

  3. Utilization of Genomic Variations Among Xylella fastidiosa Strains for Improved Diagnostic Design

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The Gram-negative, xylem-limited phytopathogenic bacterium Xylella fastidiosa causes economically important diseases in grapevine, citrus and many other plant species. Our recent whole genome comparative analysis of the four sequenced strains has identified genomic variation among these strains. The...

  4. Xylella fastidiosa infection effects on xylem sap and tissue phenolics in different grapevine cultivars

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The bacterium Xylella fastidiosa (Xf) causes Pierce’s disease (PD), a serious disease that threatens the billion-dollar California grape industry. Previous studies observed that Xf-infected grapevines, as part of a defense response, produced greater xylem levels of phenolics, which are compounds tha...

  5. Evaluation of pathogenicity and insect transmission of Xylella fastidiosa strains to olive plants

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Xylella fastidiosa (Xf) is a xylem-limited bacterium that causes disease in a number of economically important crops in California and worldwide. Newly observed scorching symptoms in olive trees may be due to Xf infection. If true, “olive leaf scorch disease” (OLSD) would represent a new threat to...

  6. Xylella fastidiosa plasmid-encoded PemK toxin is an endoribonuclease.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Stable inheritance of pXF-RIV11 in Xylella fastidiosa is conferred by the pemI/pemK plasmid addiction system. PemK serves as a toxin inhibiting bacterial growth; PemI is the corresponding antitoxin that blocks activity of PemK toxin by direct binding. PemK toxin and PemI antitoxin were over-expre...

  7. Genome-wide Profiling and Analysis of the Vitis Transcriptome Responses to Xylella fastidiosa Infection

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The gram-negative bacterium Xylella fastidiosa (Xf), vectored by xylem-feeding sharpshooters, is the causal agent of Pierce’s disease (PD) in grape. This disease causes serious economic losses in California. PD resistant plants produced by conventional or molecular breeding techniques are the ultima...

  8. Extracellular Genomic DNA Mediates Enhancement of Xylella fastidiosa Biofilm Formation in Vitro

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Xylella fastidiosa (Xf) produces extracellular DNA in PD3 liquid medium. This extracellular DNA could enhance biofilm formation, a factor in successful establishment of Xf in planta. The relative amounts of extracellular DNA were positively correlated with planktonic growth and biofilm formation in ...

  9. Grapevines undergo varying shifts in secondary metabolic profiles when infected with Xylella fastidiosa

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Pierce’s disease (PD) is a devastating disease of grapevine caused by the bacterial pathogen Xylella fastidiosa (Xf). Key to the development and optimization of PD-tolerant grape cultivars is improved understanding about how grapevines defend themselves against Xf. This study complements histologica...

  10. The Glassy-Winged Sharpshooter Vector of Xylella fastidiosa Harbors a Phytoreovirus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The glassy-winged sharpshooter (GWSS) vector of Xylella fastidiosa harbors a phytoreovirus species designated as Homalodisca vitripennis reovirus (HoVRV). Double-shelled isometric virus particles purified from GWSS adults resembled those observed in thin sections of GWSS salivary glands by transmis...

  11. Identification of Grapevine Xylem Sap Protein Profiles in Response to Xylella fastidiosa Infection

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Pierce’s Disease (PD) of grapevines is caused by the Gram-negative, xylem-limited bacterium Xylella fastidiosa (Xf). All Vitis vinifera-based cultivars are highly susceptible to Xf infection. However, some grape species from the southern United States (such as V. arizonica, V. Shuttleworthii, V. sim...

  12. Differential Susceptibility of Prunus Germplasm (Subgenus Amygdalus) to a California Strain of Xylella fastidiosa

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Seedling peach and clonal peach-almond hybrids are popular rootstock choices for commercial almond growers in California. In this study, we challenged clonal replicates of rootstock accessions of peach, almond, and a first generation peach-almond hybrid with Xylella fastidiosa biotype M23. Clonal ...

  13. Genotypic and phenotypic characterization of Xylella fastidiosa strains isolated from California, Georgia and Florida

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Xylella fastidiosa is an important plant pathogen causing Pierce’s Disease in grapes and in many other economically important crops. Because of its nutritional fastidiousness, characterization of this bacterium remains highly challenging, thereby hindering future scientific research and disease cont...

  14. Assessment of the role of alfalfa in the spread of Xylella fastidiosa in California

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Grape and almond are planted on a large scale throughout much of California. Both crops are susceptible to diseases caused by Xylella fastidiosa. Within California’s Central Valley, the green sharpshooter (Draeculacephala minerva) is a key vector. This insect is often found in large numbers in cu...

  15. Arabidopsis Thaliana Ecotypes With Differential Susceptibility To The Bacterial Pathogen Xylella fastidiosa

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Pierce’s disease of grapes and almond leaf scorch are devastating diseases caused by the bacterium Xylella fastidiosa (Xf). To date, progress in determining the mechanisms of host plant susceptibility, tolerance or resistance has been slow, due in large part to the long generation time and limited a...

  16. Draft Genome Sequence of CO33, a Coffee-Infecting Isolate of Xylella fastidiosa.

    PubMed

    Giampetruzzi, Annalisa; Loconsole, Giuliana; Boscia, Donato; Calzolari, Alessandra; Chiumenti, Michela; Martelli, Giovanni P; Saldarelli, Pasquale; Almeida, Rodrigo P P; Saponari, Maria

    2015-01-01

    The draft genome sequence of Xylella fastidiosa CO33 isolate, retrieved from symptomatic leaves of coffee plant intercepted in northern Italy, is reported. The CO33 genome size is 2,681,926 bp with a GC content of 51.7%. PMID:26679584

  17. Assessing the potential of forage alfalfa crops to serve as Xylella fastidiosa, primary inoculum sources in the San Joaquin Valley

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The potential for forage alfalfa to serve as a primary inoculum source of Xylella fastidiosa in the San Joaquin Valley of California was evaluated. Laboratory inoculation of fourteen cultivars of alfalfa indicated that all alfalfa cultivars tested were equally suitable hosts for X. fastidiosa. Inci...

  18. Interactions between Grape and Sweet Orange Strains of Xylella Fastidiosa in Colonization and Symptom Production in Periwinkle Plants

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Xylella fastidiosa is one of the most economically important bacterial plant pathogens, causing serious diseases including citrus variegated chlorosis in Brazil and Pierce’s disease of grape in California. The sweet orange strain of X. fastidiosa is a particular a threat to US citrus industry and it...

  19. Direct evidence of egestion and salivation of Xylella fastidiosa suggests sharpshooters can be “flying syringes”

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Despite nearly 70 years of research, the inoculation mechanism of Xylella fastidiosa by its sharpshooter vectors is still unproven. X. fastidiosa is unique among insect-transmitted plant pathogens because it does not circulate in the vector’s hemolymph, yet it is propagative because it adheres to an...

  20. Visualization of twitching motility and characterization of the role of the PilG in Xylella fastidiosa

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Type IV pili of Xylella fastidiosa are regulated by pilG, a chemotaxis regulator in the Pil-Chp operon involving signal transduction pathways. To elucidate the role of pilG in twitching motility and pathogenicity of X. fastidiosa, the phenotypes of wild type, a pilG-deficient mutant (Xf'pilG), and a...

  1. Functional characterization of replication and stability factors of an incP-1 plasmid from Xylella fastidiosa

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Xylella fastidiosa strain riv11 harbors a ~25 kbp plasmid (pXF-RIV11) belonging to the incP-1 incompatibility group. Replication and stability factors of pXF-RIV11 were identified and used to construct plasmids able to propagate in both X. fastidiosa and Escherichia coli. Replication in X. fastidi...

  2. Xylella fastidiosa infection of grapevines affects host secondary metabolite and defense-related protein levels within xylem

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Pierce’s disease of grapevine is a serious threat to grape production and is caused by the xylem-dwelling bacterial pathogen Xylella fastidiosa. Microscopy studies have documented morphological changes to grapevine xylem due to infection by X. fastidiosa. Comparatively, less is known about the bi...

  3. Effects of Insect Origin, Gender, and Age on Transmission of Xylella fastidiosa to Grapevines by Homalodisca vitripennis (Hemiptera: Cicadellidae)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Homalodisca vitripennis (Germar) is an invasive insect vector in California, where it transmits the bacterium Xylella fastidiosa to grapevines and other crops. Transmission efficiency of X. fastidiosa to grapevines by male and female H. vitripennis originating from two geographically separated popul...

  4. Genome-wide comparison and taxonomic relatedness of multiple Xylella fastidiosa strains reveal the occurrence of three subspecies and a new Xylella species.

    PubMed

    Marcelletti, Simone; Scortichini, Marco

    2016-10-01

    A total of 21 Xylella fastidiosa strains were assessed by comparing their genomes to infer their taxonomic relationships. The whole-genome-based average nucleotide identity and tetranucleotide frequency correlation coefficient analyses were performed. In addition, a consensus tree based on comparisons of 956 core gene families, and a genome-wide phylogenetic tree and a Neighbor-net network were constructed with 820,088 nucleotides (i.e., approximately 30-33 % of the entire X. fastidiosa genome). All approaches revealed the occurrence of three well-demarcated genetic clusters that represent X. fastidiosa subspecies fastidiosa, multiplex and pauca, with the latter appeared to diverge. We suggest that the proposed but never formally described subspecies 'sandyi' and 'morus' are instead members of the subspecies fastidiosa. These analyses support the view that the Xylella strain isolated from Pyrus pyrifolia in Taiwan is likely to be a new species. A widely used multilocus sequence typing analysis yielded conflicting results. PMID:27209415

  5. Effect of oxygen on the growth and biofilm formation of Xylella fastidiosa in liquid media.

    PubMed

    Shriner, Anthony D; Andersen, Peter C

    2014-12-01

    Xylella fastidiosa is a xylem-limited bacterial pathogen, and is the causative agent of Pierce's disease of grapevines and scorch diseases of many other plant species. The disease symptoms are putatively due to blocking of the transpiration stream by bacterial-induced biofilm formation and/or by the formation of plant-generated tylosis. Xylella fastidiosa has been classified as an obligate aerobe, which appears unusual given that dissolved O2 levels in the xylem during the growing season are often hypoxic (20-60 μmol L(-1)). We examined the growth and biofilm formation of three strains of X. fastidiosa under variable O2 conditions (21, 2.1, 0.21 and 0 % O2), in comparison to that of Pseudomonas syringae (obligate aerobe) and Erwinia carotovora (facultative anaerobe) under similar conditions. The growth of X. fastidiosa more closely resembled that of the facultative anaerobe, and not the obligate aerobe. Xanthomonas campestris, the closest genetic relative of X. fastidiosa, exhibited no growth in an N2 environment, whereas X. fastidiosa was capable of growing in an N2 environment in PW(+), CHARDS, and XDM2-PR media. The magnitude of growth and biofilm formation in the N2 (0 % O2) treatment was dependent on the specific medium. Additional studies involving the metabolism of X. fastidiosa in response to low O2 are warranted. Whether X. fastidiosa is classified as an obligate aerobe or a facultative anaerobe should be confirmed by gene activation and/or the quantification of the metabolic profiles under hypoxic conditions. PMID:25100224

  6. Comparative genomic characterization of citrus-associated Xylella fastidiosa strains

    PubMed Central

    da Silva, Vivian S; Shida, Cláudio S; Rodrigues, Fabiana B; Ribeiro, Diógenes CD; de Souza, Alessandra A; Coletta-Filho, Helvécio D; Machado, Marcos A; Nunes, Luiz R; de Oliveira, Regina Costa

    2007-01-01

    Background The xylem-inhabiting bacterium Xylella fastidiosa (Xf) is the causal agent of Pierce's disease (PD) in vineyards and citrus variegated chlorosis (CVC) in orange trees. Both of these economically-devastating diseases are caused by distinct strains of this complex group of microorganisms, which has motivated researchers to conduct extensive genomic sequencing projects with Xf strains. This sequence information, along with other molecular tools, have been used to estimate the evolutionary history of the group and provide clues to understand the capacity of Xf to infect different hosts, causing a variety of symptoms. Nonetheless, although significant amounts of information have been generated from Xf strains, a large proportion of these efforts has concentrated on the study of North American strains, limiting our understanding about the genomic composition of South American strains – which is particularly important for CVC-associated strains. Results This paper describes the first genome-wide comparison among South American Xf strains, involving 6 distinct citrus-associated bacteria. Comparative analyses performed through a microarray-based approach allowed identification and characterization of large mobile genetic elements that seem to be exclusive to South American strains. Moreover, a large-scale sequencing effort, based on Suppressive Subtraction Hybridization (SSH), identified 290 new ORFs, distributed in 135 Groups of Orthologous Elements, throughout the genomes of these bacteria. Conclusion Results from microarray-based comparisons provide further evidence concerning activity of horizontally transferred elements, reinforcing their importance as major mediators in the evolution of Xf. Moreover, the microarray-based genomic profiles showed similarity between Xf strains 9a5c and Fb7, which is unexpected, given the geographical and chronological differences associated with the isolation of these microorganisms. The newly identified ORFs, obtained by

  7. The complex biogeography of the plant pathogen Xylella fastidiosa: genetic evidence of introductions and Subspecific introgression in Central America.

    PubMed

    Nunney, Leonard; Ortiz, Beatriz; Russell, Stephanie A; Ruiz Sánchez, Rebeca; Stouthamer, Richard

    2014-01-01

    The bacterium Xylella fastidiosa is a plant pathogen with a history of economically damaging introductions of subspecies to regions where its other subspecies are native. Genetic evidence is presented demonstrating the introduction of two new taxa into Central America and their introgression into the native subspecies, X. fastidiosa subsp. fastidiosa. The data are from 10 genetic outliers detected by multilocus sequence typing (MLST) of isolates from Costa Rica. Six (five from oleander, one from coffee) defined a new sequence type (ST53) that carried alleles at six of the eight loci sequenced (five of the seven MLST loci) diagnostic of the South American subspecies Xylella fastidiosa subsp. pauca which causes two economically damaging plant diseases, citrus variegated chlorosis and coffee leaf scorch. The two remaining loci of ST53 carried alleles from what appears to be a new South American form of X. fastidiosa. Four isolates, classified as X. fastidiosa subsp. fastidiosa, showed a low level of introgression of non-native DNA. One grapevine isolate showed introgression of an allele from X. fastidiosa subsp. pauca while the other three (from citrus and coffee) showed introgression of an allele with similar ancestry to the alleles of unknown origin in ST53. The presence of X. fastidiosa subsp. pauca in Central America is troubling given its disease potential, and establishes another route for the introduction of this economically damaging subspecies into the US or elsewhere, a threat potentially compounded by the presence of a previously unknown form of X. fastidiosa. PMID:25379725

  8. Draft Genome Sequence of Xylella fastidiosa subsp. multiplex Strain Griffin-1 from Quercus rubra in Georgia.

    PubMed

    Chen, Jianchi; Huang, Hong; Chang, Chung-Jan; Stenger, Drake C

    2013-01-01

    The draft genome sequence of Xylella fastidiosa subsp. multiplex strain Griffin-1, isolated from a red oak tree (Quercus rubra) in Georgia, is reported here. The bacterium has a genome size of 2,387,314 bp, with a G+C content of 51.7%. The Griffin-1 strain genome contains 2,903 predicted open reading frames and 50 RNA genes. PMID:24115539

  9. Draft Genome Sequence of Xylella fastidiosa Pear Leaf Scorch Strain in Taiwan.

    PubMed

    Su, C-C; Deng, W-L; Jan, F-J; Chang, C-J; Huang, H; Chen, J

    2014-01-01

    The draft genome sequence of Xylella fastidiosa pear leaf scorch strain PLS229, isolated from the pear cultivar Hengshan (Pyrus pyrifolia) in Taiwan, is reported here. The bacterium has a genome size of 2,733,013 bp, with a G+C content of 53.1%. The PLS229 genome was annotated and has 3,259 open reading frames and 50 RNA genes. PMID:24652975

  10. Promiscuous Diffusible Signal Factor Production and Responsiveness of the Xylella fastidiosa Rpf System

    PubMed Central

    Ionescu, Michael; Yokota, Kenji; Antonova, Elena; Garcia, Angelica; Beaulieu, Ellen; Hayes, Terry; Iavarone, Anthony T.

    2016-01-01

    ABSTRACT Cell density-dependent regulation of gene expression in Xylella fastidiosa that is crucial to its switching between plant hosts and insect vectors is dependent on RpfF and its production of 2-enoic acids known as diffusible signal factor (DSF). We show that X. fastidiosa produces a particularly large variety of similar, relatively long-chain-length 2-enoic acids that are active in modulating gene expression. Both X. fastidiosa itself and a Pantoea agglomerans surrogate host harboring X. fastidiosa RpfF (XfRpfF) is capable of producing a variety of both saturated and unsaturated free fatty acids. However, only 2-cis unsaturated acids were found to be biologically active in X. fastidiosa. X. fastidiosa produces, and is particularly responsive to, a novel DSF species, 2-cis-hexadecanoic acid that we term XfDSF2. It is also responsive to other, even longer 2-enoic acids to which other taxa such as Xanthomonas campestris are unresponsive. The 2-enoic acids that are produced by X. fastidiosa are strongly affected by the cellular growth environment, with XfDSF2 not detected in culture media in which 2-tetradecenoic acid (XfDSF1) had previously been found. X. fastidiosa is responsive to much lower concentrations of XfDSF2 than XfDSF1. Apparently competitive interactions can occur between various saturated and unsaturated fatty acids that block the function of those agonistic 2-enoic fatty acids. By altering the particular 2-enoic acids produced and the relative balance of free enoic and saturated fatty acids, X. fastidiosa might modulate the extent of DSF-mediated quorum sensing. PMID:27435463

  11. Detection and visualization of an exopolysaccharide produced by Xylella fastidiosa in vitro and in planta.

    PubMed

    Roper, M Caroline; Greve, L Carl; Labavitch, John M; Kirkpatrick, Bruce C

    2007-11-01

    Many phytopathogenic bacteria, such as Ralstonia solanacearum, Pantoea stewartii, and Xanthomonas campestris, produce exopolysaccharides (EPSs) that aid in virulence, colonization, and survival. EPS can also contribute to host xylem vessel blockage. The genome of Xylella fastidiosa, the causal agent of Pierce's disease (PD) of grapevine, contains an operon that is strikingly similar to the X. campestris gum operon, which is responsible for the production of xanthan gum. Based on this information, it has been hypothesized that X. fastidiosa is capable of producing an EPS similar in structure and composition to xanthan gum but lacking the terminal mannose residue. In this study, we raised polyclonal antibodies against a modified xanthan gum polymer similar to the predicted X. fastidiosa EPS polymer. We used enzyme-linked immunosorbent assay to quantify production of EPS from X. fastidiosa cells grown in vitro and immunolocalization microscopy to examine the distribution of X. fastidiosa EPS in biofilms formed in vitro and in planta and assessed the contribution of X. fastidiosa EPS to the vascular occlusions seen in PD-infected grapevines. PMID:17827325

  12. The Secreted Protease PrtA Controls Cell Growth, Biofilm Formation and Pathogenicity in Xylella fastidiosa

    PubMed Central

    Gouran, Hossein; Gillespie, Hyrum; Nascimento, Rafael; Chakraborty, Sandeep; Zaini, Paulo A.; Jacobson, Aaron; Phinney, Brett S.; Dolan, David; Durbin-Johnson, Blythe P.; Antonova, Elena S.; Lindow, Steven E.; Mellema, Matthew S.; Goulart, Luiz R.; Dandekar, Abhaya M.

    2016-01-01

    Pierce’s disease (PD) is a deadly disease of grapevines caused by the Gram-negative bacterium Xylella fastidiosa. Though disease symptoms were formerly attributed to bacteria blocking the plant xylem, this hypothesis is at best overly simplistic. Recently, we used a proteomic approach to characterize the secretome of X. fastidiosa, both in vitro and in planta, and identified LesA as one of the pathogenicity factors of X. fastidiosa in grapevines that leads to leaf scorching and chlorosis. Herein, we characterize another such factor encoded by PD0956, designated as an antivirulence secreted protease “PrtA” that displays a central role in controlling in vitro cell proliferation, length, motility, biofilm formation, and in planta virulence. The mutant in X. fastidiosa exhibited reduced cell length, hypermotility (and subsequent lack of biofilm formation) and hypervirulence in grapevines. These findings are supported by transcriptomic and proteomic analyses with corresponding plant infection data. Of particular interest, is the hypervirulent response in grapevines observed when X. fastidiosa is disrupted for production of PrtA, and that PD-model tobacco plants transformed to express PrtA exhibited decreased symptoms after infection by X. fastidiosa. PMID:27492542

  13. The Secreted Protease PrtA Controls Cell Growth, Biofilm Formation and Pathogenicity in Xylella fastidiosa.

    PubMed

    Gouran, Hossein; Gillespie, Hyrum; Nascimento, Rafael; Chakraborty, Sandeep; Zaini, Paulo A; Jacobson, Aaron; Phinney, Brett S; Dolan, David; Durbin-Johnson, Blythe P; Antonova, Elena S; Lindow, Steven E; Mellema, Matthew S; Goulart, Luiz R; Dandekar, Abhaya M

    2016-01-01

    Pierce's disease (PD) is a deadly disease of grapevines caused by the Gram-negative bacterium Xylella fastidiosa. Though disease symptoms were formerly attributed to bacteria blocking the plant xylem, this hypothesis is at best overly simplistic. Recently, we used a proteomic approach to characterize the secretome of X. fastidiosa, both in vitro and in planta, and identified LesA as one of the pathogenicity factors of X. fastidiosa in grapevines that leads to leaf scorching and chlorosis. Herein, we characterize another such factor encoded by PD0956, designated as an antivirulence secreted protease "PrtA" that displays a central role in controlling in vitro cell proliferation, length, motility, biofilm formation, and in planta virulence. The mutant in X. fastidiosa exhibited reduced cell length, hypermotility (and subsequent lack of biofilm formation) and hypervirulence in grapevines. These findings are supported by transcriptomic and proteomic analyses with corresponding plant infection data. Of particular interest, is the hypervirulent response in grapevines observed when X. fastidiosa is disrupted for production of PrtA, and that PD-model tobacco plants transformed to express PrtA exhibited decreased symptoms after infection by X. fastidiosa. PMID:27492542

  14. Direct Evidence of Egestion and Salivation of Xylella fastidiosa Suggests Sharpshooters Can Be "Flying Syringes".

    PubMed

    Backus, Elaine A; Shugart, Holly J; Rogers, Elizabeth E; Morgan, J Kent; Shatters, Robert

    2015-05-01

    Xylella fastidiosa is unique among insect-transmitted plant pathogens because it is propagative but noncirculative, adhering to and multiplying on the cuticular lining of the anterior foregut. Any inoculation mechanism for X. fastidiosa must explain how bacterial cells exit the vector's stylets via the food canal and directly enter the plant. A combined egestion-salivation mechanism has been proposed to explain these unique features. Egestion is the putative outward flow of fluid from the foregut via hypothesized bidirectional pumping of the cibarium. The present study traced green fluorescent protein-expressing X. fastidiosa or fluorescent nanoparticles acquired from artificial diets by glassy-winged sharpshooters, Homalodisca vitripennis, as they were egested into simultaneously secreted saliva. X. fastidiosa or nanoparticles were shown to mix with gelling saliva to form fluorescent deposits and salivary sheaths on artificial diets, providing the first direct, conclusive evidence of egestion by any hemipteran insect. Therefore, the present results strongly support an egestion-salivation mechanism of X. fastidiosa inoculation. Results also support that a column of fluid is transiently held in the foregut without being swallowed. Evidence also supports (but does not definitively prove) that bacteria were suspended in the column of fluid during the vector's transit from diet to diet, and were egested with the held fluid. Thus, we hypothesize that sharpshooters could be true "flying syringes," especially when inoculation occurs very soon after uptake of bacteria, suggesting the new paradigm of a nonpersistent X. fastidiosa transmission mechanism. PMID:26020829

  15. New Coffee Plant-Infecting Xylella fastidiosa Variants Derived via Homologous Recombination.

    PubMed

    Jacques, Marie-Agnès; Denancé, Nicolas; Legendre, Bruno; Morel, Emmanuelle; Briand, Martial; Mississipi, Stelly; Durand, Karine; Olivier, Valérie; Portier, Perrine; Poliakoff, Françoise; Crouzillat, Dominique

    2015-01-01

    Xylella fastidiosa is a xylem-limited phytopathogenic bacterium endemic to the Americas that has recently emerged in Asia and Europe. Although this bacterium is classified as a quarantine organism in the European Union, importation of plant material from contaminated areas and latent infection in asymptomatic plants have engendered its inevitable introduction. In 2012, four coffee plants (Coffea arabica and Coffea canephora) with leaf scorch symptoms growing in a confined greenhouse were detected and intercepted in France. After identification of the causal agent, this outbreak was eradicated. Three X. fastidiosa strains were isolated from these plants, confirming a preliminary identification based on immunology. The strains were characterized by multiplex PCR and by multilocus sequence analysis/typing (MLSA-MLST) based on seven housekeeping genes. One strain, CFBP 8073, isolated from C. canephora imported from Mexico, was assigned to X. fastidiosa subsp. fastidiosa/X. fastidiosa subsp. sandyi. This strain harbors a novel sequence type (ST) with novel alleles at two loci. The two other strains, CFBP 8072 and CFBP 8074, isolated from Coffea arabica imported from Ecuador, were allocated to X. fastidiosa subsp. pauca. These two strains shared a novel ST with novel alleles at two loci. These MLST profiles showed evidence of recombination events. We provide genome sequences for CFBP 8072 and CFBP 8073 strains. Comparative genomic analyses of these two genome sequences with publicly available X. fastidiosa genomes, including the Italian strain CoDiRO, confirmed these phylogenetic positions and provided candidate alleles for coffee plant adaptation. This study demonstrates the global diversity of X. fastidiosa and highlights the diversity of strains isolated from coffee plants. PMID:26712553

  16. Radicinin from Cochliobolus sp. inhibits Xylella fastidiosa, the causal agent of Pierce's Disease of grapevine.

    PubMed

    Aldrich, Thomas J; Rolshausen, Philippe E; Roper, M Caroline; Reader, Jordan M; Steinhaus, Matthew J; Rapicavoli, Jeannette; Vosburg, David A; Maloney, Katherine N

    2015-08-01

    The fastidious phytopathogenic bacterium, Xylella fastidiosa, poses a substantial threat to many economically important crops, causing devastating diseases including Pierce's Disease of grapevine. Grapevines (Vitis vinifera L.) planted in an area under Pierce's Disease pressure often display differences in disease severity and symptom expression, with apparently healthy vines growing alongside the dying ones, despite the fact that all the vines are genetic clones of one another. Under the hypothesis that endophytic microbes might be responsible for this non-genetic resistance to X. fastidiosa, endophytic fungi were isolated from vineyard cvs. 'Chardonnay' and 'Cabernet Sauvignon' grown under high Pierce's Disease pressure. A Cochliobolus sp. isolated from a Cabernet Sauvignon grapevine inhibited the growth of X. fastidiosa in vitro. Bioassay-guided isolation of an organic extract of Cochliobolus sp. yielded the natural product radicinin as the major active compound. Radicinin also inhibited proteases isolated from the culture supernatant of X. fastidiosa. In order to assess structure-activity relationships, three semi-synthetic derivatives of radicinin were prepared and tested for activity against X. fastidiosa in vitro. Assay results of these derivatives are consistent with enzyme inactivation by conjugate addition to carbon-10 of radicinin, as proposed previously. PMID:25892412

  17. Characterization of regulatory pathways in Xylella fastidiosa: genes and phenotypes controlled by gacA.

    PubMed

    Shi, Xiang Yang; Dumenyo, C Korsi; Hernandez-Martinez, Rufina; Azad, Hamid; Cooksey, Donald A

    2009-04-01

    The xylem-limited, insect-transmitted bacterium Xylella fastidiosa causes Pierce's disease in grapes through cell aggregation and vascular clogging. GacA controls various physiological processes and pathogenicity factors in many gram-negative bacteria, including biofilm formation in Pseudomonas syringae pv. tomato DC3000. Cloned gacA of X. fastidiosa was found to restore the hypersensitive response and pathogenicity in gacA mutants of P. syringae pv. tomato DC3000 and Erwinia amylovora. A gacA mutant of X. fastidiosa (DAC1984) had significantly reduced abilities to adhere to a glass surface, form biofilm, and incite disease symptoms on grapevines, compared with the parent (A05). cDNA microarray analysis identified 7 genes that were positively regulated by GacA, including xadA and hsf, predicted to encode outer membrane adhesion proteins, and 20 negatively regulated genes, including gumC and an antibacterial polypeptide toxin gene, cvaC. These results suggest that GacA of X. fastidiosa regulates many factors, which contribute to attachment and biofilm formation, as well as some physiological processes that may enhance the adaptation and tolerance of X. fastidiosa to environmental stresses and the competition within the host xylem. PMID:19218414

  18. Comparative genomic analysis of coffee-infecting Xylella fastidiosa strains isolated from Brazil.

    PubMed

    Barbosa, Deibs; Alencar, Valquíria Campos; Santos, Daiene Souza; de Freitas Oliveira, Ana Cláudia; de Souza, Alessandra A; Coletta-Filho, Helvecio D; de Oliveira, Regina Souza; Nunes, Luiz R

    2015-05-01

    Strains of Xylella fastidiosa constitute a complex group of bacteria that develop within the xylem of many plant hosts, causing diseases of significant economic importance, such as Pierce's disease in North American grapevines and citrus variegated chlorosis in Brazil. X. fastidiosa has also been obtained from other host plants, in direct correlation with the development of diseases, as in the case of coffee leaf scorch (CLS)--a disease with potential to cause severe economic losses to the Brazilian coffee industry. This paper describes a thorough genomic characterization of coffee-infecting X. fastidiosa strains, initially performed through a microarray-based approach, which demonstrated that CLS strains could be subdivided in two phylogenetically distinct subgroups. Whole-genomic sequencing of two of these bacteria (one from each subgroup) allowed identification of ORFs and horizontally transferred elements (HTEs) that were specific to CLS-related X. fastidiosa strains. Such analyses confirmed the size and importance of HTEs as major mediators of chromosomal evolution amongst these bacteria, and allowed identification of differences in gene content, after comparisons were made with previously sequenced X. fastidiosa strains, isolated from alternative hosts. Although direct experimentation still needs to be performed to elucidate the biological consequences associated with such differences, it was interesting to verify that CLS-related bacteria display variations in genes that produce toxins, as well as surface-related factors (such as fimbrial adhesins and LPS) that have been shown to be involved with recognition of specific host factors in different pathogenic bacteria. PMID:25737482

  19. Differentiation of Xylella fastidiosa Strains via Multilocus Sequence Analysis of Environmentally Mediated Genes (MLSA-E)

    PubMed Central

    Parker, Jennifer K.; Havird, Justin C.

    2012-01-01

    Isolates of the plant pathogen Xylella fastidiosa are genetically very similar, but studies on their biological traits have indicated differences in virulence and infection symptomatology. Taxonomic analyses have identified several subspecies, and phylogenetic analyses of housekeeping genes have shown broad host-based genetic differences; however, results are still inconclusive for genetic differentiation of isolates within subspecies. This study employs multilocus sequence analysis of environmentally mediated genes (MLSA-E; genes influenced by environmental factors) to investigate X. fastidiosa relationships and differentiate isolates with low genetic variability. Potential environmentally mediated genes, including host colonization and survival genes related to infection establishment, were identified a priori. The ratio of the rate of nonsynonymous substitutions to the rate of synonymous substitutions (dN/dS) was calculated to select genes that may be under increased positive selection compared to previously studied housekeeping genes. Nine genes were sequenced from 54 X. fastidiosa isolates infecting different host plants across the United States. Results of maximum likelihood (ML) and Bayesian phylogenetic (BP) analyses are in agreement with known X. fastidiosa subspecies clades but show novel within-subspecies differentiation, including geographic differentiation, and provide additional information regarding host-based isolate variation and specificity. dN/dS ratios of environmentally mediated genes, though <1 due to high sequence similarity, are significantly greater than housekeeping gene dN/dS ratios and correlate with increased sequence variability. MLSA-E can more precisely resolve relationships between closely related bacterial strains with low genetic variability, such as X. fastidiosa isolates. Discovering the genetic relationships between X. fastidiosa isolates will provide new insights into the epidemiology of populations of X. fastidiosa, allowing

  20. Multiplexed lateral flow microarray assay for detection of citrus pathogens Xylella fastidiosa and Xanthomonas axonopodis pv citri

    DOEpatents

    Cary; R. Bruce; Stubben, Christopher J.

    2011-03-22

    The invention provides highly sensitive and specific assays for the major citrus pathogens Xylella fastidiosa and Xanthomonas axonopodis, including a field deployable multiplexed assay capable of rapidly assaying for both pathogens simultaneously. The assays are directed at particular gene targets derived from pathogenic strains that specifically cause the major citrus diseases of citrus variegated chlorosis (Xylella fastidiosa 9a5c) and citrus canker (Xanthomonas axonopodis pv citri). The citrus pathogen assays of the invention offer femtomole sensitivity, excellent linear dynamic range, and rapid and specific detection.

  1. Colony aggregation and biofilm formation in xylem chemistry-based media for Xylella fastidiosa.

    PubMed

    Leite, Breno; Andersen, Peter Craig; Ishida, Maria Lucia

    2004-01-30

    Two chemically defined media based on xylem fluid chemistry were developed for Xylella fastidiosa. These media were tested and compared to chemically defined media XDM2, XDM4 and XF-26. New media were evaluated for the Pierce's disease (PD) strain UCLA-PD. Our media either was similar to the concentration of some amino acids found in the xylem fluid of the PD-susceptible Vitis vinifera cv. Chardonnay (medium CHARD2) or incorporated the tripeptide glutathione found in xylem fluid composition (medium 3G10-R). CHARD2 and 3G10-R are among the simplest chemically defined media available. Xylem fluid chemistry-based media supported X. fastidiosa growth and especially stimulated aggregation and biofilm formation. PMID:14757251

  2. Origins of the Xylella fastidiosa Prophage-Like Regions and Their Impact in Genome Differentiation

    PubMed Central

    de Mello Varani, Alessandro; Souza, Rangel Celso; Nakaya, Helder I.; de Lima, Wanessa Cristina; Paula de Almeida, Luiz Gonzaga; Kitajima, Elliot Watanabe; Chen, Jianchi; Civerolo, Edwin; Vasconcelos, Ana Tereza Ribeiro; Van Sluys, Marie-Anne

    2008-01-01

    Xylella fastidiosa is a Gram negative plant pathogen causing many economically important diseases, and analyses of completely sequenced X. fastidiosa genome strains allowed the identification of many prophage-like elements and possibly phage remnants, accounting for up to 15% of the genome composition. To better evaluate the recent evolution of the X. fastidiosa chromosome backbone among distinct pathovars, the number and location of prophage-like regions on two finished genomes (9a5c and Temecula1), and in two candidate molecules (Ann1 and Dixon) were assessed. Based on comparative best bidirectional hit analyses, the majority (51%) of the predicted genes in the X. fastidiosa prophage-like regions are related to structural phage genes belonging to the Siphoviridae family. Electron micrograph reveals the existence of putative viral particles with similar morphology to lambda phages in the bacterial cell in planta. Moreover, analysis of microarray data indicates that 9a5c strain cultivated under stress conditions presents enhanced expression of phage anti-repressor genes, suggesting switches from lysogenic to lytic cycle of phages under stress-induced situations. Furthermore, virulence-associated proteins and toxins are found within these prophage-like elements, thus suggesting an important role in host adaptation. Finally, clustering analyses of phage integrase genes based on multiple alignment patterns reveal they group in five lineages, all possessing a tyrosine recombinase catalytic domain, and phylogenetically close to other integrases found in phages that are genetic mosaics and able to perform generalized and specialized transduction. Integration sites and tRNA association is also evidenced. In summary, we present comparative and experimental evidence supporting the association and contribution of phage activity on the differentiation of Xylella genomes. PMID:19116666

  3. Grapevine rootstock effects on scion sap phenolic levels, resistance to Xylella fastidiosa infection, and progression of Pierce’s disease

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The xylem-limited bacterium Xylella fastidiosa (Xf) causes Pierce’s disease (PD), an important disease of grapevine, Vitis vinifera L. Grapevine rootstocks were developed to provide increased resistance to root disease, but rootstock effects on cane and vine diseases remain unclear. Grapevines that ...

  4. Genetic structure of Xylella fastidiosa within two important grape growing regions in the United States: California and Texas

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Xylella fastidiosa (Xf) causes Pierce’s disease in grapevine. Here, we report on the genetic diversity and population genetic structure of grape Xf strains between two important grape growing regions in the United States, California and Texas. Using multilocus microsatellite markers, high genetic di...

  5. The transmission mechanism of Xylella fastidiosa by sharpshooter vectors and a model for quantification of vector efficiency

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The exact mechanism of transmission (acquisition and inoculation) of Xylella fastidiosa (Xf) by sharpshooter vectors such as the glassy-winged sharpshooter, Homalodisca vitripennis (Germar), has been unknown. This presentation will summarize five years of work to identify the transmission mechanism...

  6. How to be an ideal vector: Four crucial steps in the transmission mechanism of Xylella fastidiosa by sharpshooters

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This paper discusses recent research on how the feeding of glassy-winged sharpshooter, Homalodisca vitripennis controls transmission (acquisition and inoculation) of Xylella fastidiosa. Studies to date support a model of four crucial steps that must all occur simultaneously during H. vitripennis fe...

  7. Sharpshooter feeding behavior in relation to transmission of Xylella fastidiosa: A model for foregut-borne transmission mechanisms

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Xylella fastidiosa is a xylem-limited bacterium that causes several scorch diseases in important crops, such as Pierce’s disease of grape. Bacteria form a dense biofilm on the foregut cuticle of the glassy-winged sharpshooter, H. vitripennis, and other xylem-feeding vectors. Bacteria are inoculate...

  8. Support for the salivation-egestion hypothesis for Xylella fastidiosa inoculation: X-ray studies supporting the existence of egestion.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The mechanism of inoculation of the Pierce’s disease bacterium, Xylella fastidiosa (Xf), by vectors such as the glassy-winged sharpshooter (GWSS) is still unknown, despite nearly 70 years of study. Research in support of the salivation-egestion hypothesis for Xf inoculation is presented. Two impor...

  9. Effects of plant water stress and temperature on glassy-winged sharpshooter feeding in relation to transmission of Xylella fastidiosa

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The glassy-winged sharpshooter (GWSS), Homalodisca vitripennis (Germar), is an economically important pest of grapevine, stone fruits, nursery trees, and ornamental plants in California because it transmits Xylella fastidiosa (Xf). Two related studies examined whether GWSS feeding behaviors that co...

  10. Effects of plant water stress on vector feeding behaviors that control acquisition and inoculation of Xylella fastidiosa

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Xylella fastidiosa (Xf) is an economically important pathogen of grapevine (Pierce’s disease), stone fruits, nursery trees, and ornamental plants (various scorch diseases) in California. The bacterium is transmitted by sharpshooter leafhopper vectors, such as the glassy-winged sharpshooter (GWSS),...

  11. Plasmid vectors for Xylella fastidiosa utilizing a toxin-antitoxin system for plasmid stability in the absence of antibiotic selection

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The phytopathogen Xylella fastidiosa causes disease in a variety of important crop and landscape plants. Functional genetic studies have led to a broader understanding of virulence mechanisms used by this pathogen in the grapevine host. Plasmid shuttle vectors are important tools in studies of bacte...

  12. Support for the salivation-egestion hypothesis for Xylella fastidiosa inoculation: salivary glucanase is injected into xylem during vector feeding.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The salivation-egestion hypothesis for the inoculation mechanism of Xylella fastidiosa (Xf) proposes that saliva secreted into plants is taken up into the vector’s precibarium. There, saliva loosens the Xf bacterial biofilm by enzymatically degrading ß-1, 4 glucans that form the chemical backbone o...

  13. Support for the salivation-egestion hypothesis for Xylella fastidiosa inoculation: bacterial cells can penetrate vector saliva in xylem

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Research is underway to develop varieties of grape that are resistant to Pierce’s disease (PD) caused by Xylella fastidiosa (Xf). PD has become economically important since introduction of an exotic vector, glassy-winged sharpshooter (GWSS). However, achieving resistance to vector acquisition and ...

  14. Characterization of Xylella fastidiosa lipopolysaccharide and its role in key steps of the disease cycle in grapevine

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This project aims to elucidate molecular mechanisms of Xylella fastidiosa (Xf) pathogenicity. Work is focused on the lipopolysaccharide (LPS) component of the outer membrane, which consists of lipid A, core oligosaccharides, and a variable O-antigen moiety. Specifically, the O-antigen portion of LPS...

  15. Characterization of Xylella fastidiosa lipopolysaccharide and its role in key steps of the disease cycle in grapevine

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Xylella fastidiosa (Xf) is the causal agent of Pierce’s Disease of grapevine (PD), and also colonizes and causes disease in other crops such as almond, citrus, and oleander. While all identified Xf isolates belong to the same species, some isolates can cause disease in one host, but not another. T...

  16. Epidemiology of Diseases Caused by Xylella fastidiosa in California: Evaluation of Alfalfa As A Source of Vectors and Inocula

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Pierce’s disease of grape and almond leaf scorch disease have been chronic problems in California. Both diseases are caused by the xylem-limited, bacterial pathogen, Xylella fastidiosa. The pathogen is vectored by xylem feeding insects and within California key vectors vary by region. The potenti...

  17. Xylella fastidiosa in rabbiteye blueberry in Louisiana is genetically similar to a strain found in Southern highbush blueberry in Georgia

    Technology Transfer Automated Retrieval System (TEKTRAN)

    During the past ten years, Xylella fastidiosa has been confirmed as a pathogen of Southern highbush blueberry (Vaccinium corymbosum interspecific hybrids) in Georgia and Florida. Recent work in Louisiana has shown that it is also associated with reduced yield and altered fruit quality in ‘Tifblue’ ...

  18. CHARACTERIZATION AND IDENTIFICATION OF PD RESISTANCE MECHANISMS: ANALYSES OF XYLEM ANATOMICAL STRUCTURES AND MOLECULAR INTERACTIONS OF HOST/XYLELLA FASTIDIOSA

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The xylem-limited bacterium Xylella fastidiosa (Xf) causes Pierce’s disease (PD) with symptoms primarily due to xylem vessel blockage in susceptible grapevines. Virulence may be related to pathogen titer, presence or absence of chemical compounds in xylem sap, and/or anatomical features of the xylem...

  19. Two whole genome sequences of Xylella fastidiosa (strains M12 and M23) causing almond leaf scorch disease in California

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Xylella fastidiosa is a Gram negative, nutritionally fastidious plant pathogenic bacterium that causes many economically important diseases including almond leaf scorch disease (ALSD) and Pierce’s disease of grape in California, as well as citrus variegated chlorosis in South America. Genome inform...

  20. Seasonal fluctuations of sap-feeding insect species infected by Xylella fastidiosa in apulian olive groves of southern Italy

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A study on seasonal abundance and infectivity by Xylella fastidiosa of Auchenorrhyncha species in the Apulia region of Italy was conducted to identify ideal periods for monitoring and adoption of potential control measures against insect vectors. Adult populations of Auchenorrhyncha species were mon...

  1. The Egestion-Salivation Hypothesis: Evidence for the role of vector saliva in the inoculation mechanism of Xylella fastidiosa

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Despite ca. 40 years of study, the mechanism of inoculation of the Pierce’s Disease bacterium, Xylella fastidiosa (Xf), by vectors such as the glassy-winged sharpshooter (GWSS) is still unknown. Research on the Egestion-Salivation Hypothesis for Xf inoculation will be presented. Two important step...

  2. Population structure of Xylella fastidiosa associated with almond leaf scorch disease in the San Joaquin Valley of California

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Xylella fastidiosa (Xf) causes disease in many commercial crops including almond leaf scorch (ALS) disease in susceptible almond (Prunus dulcis). In this study, genetic diversity and population structure of Xf associated with ALS disease were evaluated. Strains isolated from two almond production si...

  3. Seasonal abundance of Draeculacephala minerva and other Xylella fastidiosa vectors in California almond orchards and vineyards.

    PubMed

    Daane, Kent M; Wistrom, Christina M; Shapland, Elaine B; Sisterson, Mark S

    2011-04-01

    Almond leaf scorch (ALS) disease is caused by the bacterium Xylella fastidiosa and transmitted by xylem-feeding insects. Reports of increased incidence of ALS-diseased trees in California prompted surveys in three almond [Prunus dulcis (Mill.) D. A. Webb]-growing regions, from June 2003 to September 2005, to determine insect vector species composition and abundance. For comparison, sampling in and near vineyards in the San Joaquin Valley, California, also was completed. Sampling in or near almond orchards collected >42,000 Cicadomorpha of which 4.8% were xylem feeders, including 1912 grass sharpshooter, Draeculacephala minerva Ball; five Xyphon fulgida Nottingham; and a single spittlebug, Philaenus spumarius L. The most abundant vector was D. minerva. Season-long sampling indicated that D. minerva was a year-round resident in and/or near almonds in the Sacramento Valley, but not in the San Joaquin Valley. Similarly, D. minerca was rare in vineyards in the San Joaquin Valley, but was abundant in irrigated pastures near vineyards. D. minerva was most frequently collected along orchard margins, and peak densities were observed in summer, the period of time when bacterial titers are reported to increase in infected trees. Screening of D. minerva for presence of X.fastidiosa found that 1.1% of insects collected near almond orchards and 4.5% of insects collected from pastures tested positive. The X. fastidiosa subspecies and genotype detected in insects collected from orchards matched those collected from ALS-diseased almond trees in the same orchard. Of the few X. fulgida and P. spumarius collected, none tested positive for X. fastidiosa. Results are discussed with respect to X. fastidiosa vector control and detection methods. PMID:21510181

  4. Three New Pierce's Disease Pathogenicity Effectors Identified Using Xylella fastidiosa Biocontrol Strain EB92-1.

    PubMed

    Zhang, Shujian; Chakrabarty, Pranjib K; Fleites, Laura A; Rayside, Patricia A; Hopkins, Donald L; Gabriel, Dean W

    2015-01-01

    Xylella fastidiosa (X. fastidiosa) infects a wide range of plant hosts and causes economically serious diseases, including Pierce's Disease (PD) of grapevines. X. fastidiosa biocontrol strain EB92-1 was isolated from elderberry and is infectious and persistent in grapevines but causes only very slight symptoms under ideal conditions. The draft genome of EB92-1 revealed that it appeared to be missing genes encoding 10 potential PD pathogenicity effectors found in Temecula1. Subsequent PCR and sequencing analyses confirmed that EB92-1 was missing the following predicted effectors found in Temecula1: two type II secreted enzymes, including a lipase (LipA; PD1703) and a serine protease (PD0956); two identical genes encoding proteins similar to Zonula occludens toxins (Zot; PD0915 and PD0928), and at least one relatively short, hemagglutinin-like protein (PD0986). Leaves of tobacco and citrus inoculated with cell-free, crude protein extracts of E. coli BL21(DE3) overexpressing PD1703 exhibited a hypersensitive response (HR) in less than 24 hours. When cloned into shuttle vector pBBR1MCS-5, PD1703 conferred strong secreted lipase activity to Xanthomonas citri, E. coli and X. fastidiosa EB92-1 in plate assays. EB92-1/PD1703 transformants also showed significantly increased disease symptoms on grapevines, characteristic of PD. Genes predicted to encode PD0928 (Zot) and a PD0986 (hemagglutinin) were also cloned into pBBR1MCS-5 and moved into EB92-1; both transformants also showed significantly increased symptoms on V. vinifera vines, characteristic of PD. Together, these results reveal that PD effectors include at least a lipase, two Zot-like toxins and a possibly redundant hemagglutinin, none of which are necessary for parasitic survival of X. fastidiosa populations in grapevines or elderberry. PMID:26218423

  5. Characterization of Novel Virulent Broad-Host-Range Phages of Xylella fastidiosa and Xanthomonas

    PubMed Central

    Ahern, Stephen J.; Das, Mayukh; Bhowmick, Tushar Suvra; Young, Ry

    2014-01-01

    The xylem-limited bacterium Xylella fastidiosa is the causal agent of several plant diseases, most notably Pierce's disease of grape and citrus variegated chlorosis. We report the isolation and characterization of the first virulent phages for X. fastidiosa, siphophages Sano and Salvo and podophages Prado and Paz, with a host range that includes Xanthomonas spp. Phages propagated on homologous hosts had observed adsorption rate constants of ∼4 × 10−12 ml cell−1 min−1 for X. fastidiosa strain Temecula 1 and ∼5 × 10−10 to 7 × 10−10 ml cell−1 min−1 for Xanthomonas strain EC-12. Sano and Salvo exhibit >80% nucleotide identity to each other in aligned regions and are syntenic to phage BcepNazgul. We propose that phage BcepNazgul is the founding member of a novel phage type, to which Sano and Salvo belong. The lysis genes of the Nazgul-like phage type include a gene that encodes an outer membrane lipoprotein endolysin and also spanin gene families that provide insight into the evolution of the lysis pathway for phages of Gram-negative hosts. Prado and Paz, although exhibiting no significant DNA homology to each other, are new members of the phiKMV-like phage type, based on the position of the single-subunit RNA polymerase gene. The four phages are type IV pilus dependent for infection of both X. fastidiosa and Xanthomonas. The phages may be useful as agents for an effective and environmentally responsible strategy for the control of diseases caused by X. fastidiosa. PMID:24214944

  6. Characterization of novel virulent broad-host-range phages of Xylella fastidiosa and Xanthomonas.

    PubMed

    Ahern, Stephen J; Das, Mayukh; Bhowmick, Tushar Suvra; Young, Ry; Gonzalez, Carlos F

    2014-01-01

    The xylem-limited bacterium Xylella fastidiosa is the causal agent of several plant diseases, most notably Pierce's disease of grape and citrus variegated chlorosis. We report the isolation and characterization of the first virulent phages for X. fastidiosa, siphophages Sano and Salvo and podophages Prado and Paz, with a host range that includes Xanthomonas spp. Phages propagated on homologous hosts had observed adsorption rate constants of ~4 × 10(-12) ml cell(-1) min(-1) for X. fastidiosa strain Temecula 1 and ~5 × 10(-10) to 7 × 10(-10) ml cell(-1) min(-1) for Xanthomonas strain EC-12. Sano and Salvo exhibit >80% nucleotide identity to each other in aligned regions and are syntenic to phage BcepNazgul. We propose that phage BcepNazgul is the founding member of a novel phage type, to which Sano and Salvo belong. The lysis genes of the Nazgul-like phage type include a gene that encodes an outer membrane lipoprotein endolysin and also spanin gene families that provide insight into the evolution of the lysis pathway for phages of Gram-negative hosts. Prado and Paz, although exhibiting no significant DNA homology to each other, are new members of the phiKMV-like phage type, based on the position of the single-subunit RNA polymerase gene. The four phages are type IV pilus dependent for infection of both X. fastidiosa and Xanthomonas. The phages may be useful as agents for an effective and environmentally responsible strategy for the control of diseases caused by X. fastidiosa. PMID:24214944

  7. Three New Pierce's Disease Pathogenicity Effectors Identified Using Xylella fastidiosa Biocontrol Strain EB92-1

    PubMed Central

    Zhang, Shujian; Chakrabarty, Pranjib K.; Fleites, Laura A.; Rayside, Patricia A.; Hopkins, Donald L.; Gabriel, Dean W.

    2015-01-01

    Xylella fastidiosa (X. fastidiosa) infects a wide range of plant hosts and causes economically serious diseases, including Pierce's Disease (PD) of grapevines. X. fastidiosa biocontrol strain EB92-1 was isolated from elderberry and is infectious and persistent in grapevines but causes only very slight symptoms under ideal conditions. The draft genome of EB92-1 revealed that it appeared to be missing genes encoding 10 potential PD pathogenicity effectors found in Temecula1. Subsequent PCR and sequencing analyses confirmed that EB92-1 was missing the following predicted effectors found in Temecula1: two type II secreted enzymes, including a lipase (LipA; PD1703) and a serine protease (PD0956); two identical genes encoding proteins similar to Zonula occludens toxins (Zot; PD0915 and PD0928), and at least one relatively short, hemagglutinin-like protein (PD0986). Leaves of tobacco and citrus inoculated with cell-free, crude protein extracts of E. coli BL21(DE3) overexpressing PD1703 exhibited a hypersensitive response (HR) in less than 24 hours. When cloned into shuttle vector pBBR1MCS-5, PD1703 conferred strong secreted lipase activity to Xanthomonas citri, E. coli and X. fastidiosa EB92-1 in plate assays. EB92-1/PD1703 transformants also showed significantly increased disease symptoms on grapevines, characteristic of PD. Genes predicted to encode PD0928 (Zot) and a PD0986 (hemagglutinin) were also cloned into pBBR1MCS-5 and moved into EB92-1; both transformants also showed significantly increased symptoms on V. vinifera vines, characteristic of PD. Together, these results reveal that PD effectors include at least a lipase, two Zot-like toxins and a possibly redundant hemagglutinin, none of which are necessary for parasitic survival of X. fastidiosa populations in grapevines or elderberry. PMID:26218423

  8. Draft Genome Sequence of Xylella fastidiosa subsp. fastidiosa Strain Stag's Leap.

    PubMed

    Chen, J; Wu, F; Zheng, Z; Deng, X; Burbank, L P; Stenger, D C

    2016-01-01

    ITALIC! Xylella fastidiosasubsp. ITALIC! fastidiosacauses Pierce's disease of grapevine. Presented here is the draft genome sequence of the Stag's Leap strain, previously used in pathogenicity/virulence assays to evaluate grapevine germplasm bearing Pierce's disease resistance and a phenotypic assessment of knockout mutants to determine gene function. PMID:27103713

  9. Water deficit modulates the response of Vitis vinifera to the Pierce's disease pathogen Xylella fastidiosa.

    PubMed

    Choi, Hong-Kyu; Iandolino, Alberto; da Silva, Francisco Goes; Cook, Douglas R

    2013-06-01

    Pierce's disease, caused by the bacterium Xylella fastidiosa, is one of the most devastating diseases of cultivated grape, currently restricted to the Americas. To test the long-standing hypothesis that Pierce's disease results from pathogen-induced drought stress, we used the Affymetrix Vitis GeneChip to compare the transcriptional response of Vitis vinifera to Xylella infection, water deficit, or a combination of the two stresses. The results reveal a redirection of gene transcription involving 822 genes with a minimum twofold change (P < 0.05), including the upregulation of transcripts for phenylpropanoid and flavonoid biosynthesis, pathogenesis-related proteins, abscisic acid- and jasmonic acid-responsive biosynthesis, and downregulation of transcripts related to photosynthesis, growth, and nutrition. Although the transcriptional response of plants to Xylella infection was largely distinct from the response of healthy plants to water stress, we find that 138 of the pathogen-induced genes exhibited a significantly stronger transcriptional response when plants were simultaneously exposed to infection and drought stress, suggesting a strong interaction between disease and water deficit. This interaction between drought stress and disease was mirrored in planta at the physiological level for aspects of water relations and photosynthesis and in terms of the severity of disease symptoms and the extent of pathogen colonization, providing a molecular correlate of the classical concept of the disease triangle in which environment impacts disease severity. PMID:23425100

  10. VapD in Xylella fastidiosa Is a Thermostable Protein with Ribonuclease Activity

    PubMed Central

    Mendes, Juliano S.; Santiago, André da S.; Toledo, Marcelo A. S.; Rosselli-Murai, Luciana K.; Favaro, Marianna T. P.; Santos, Clelton A.; Horta, Maria Augusta C.; Crucello, Aline; Beloti, Lilian L.; Romero, Fabian; Tasic, Ljubica; de Souza, Alessandra A.; de Souza, Anete P.

    2015-01-01

    Xylella fastidiosa strain 9a5c is a gram-negative phytopathogen that is the causal agent of citrus variegated chlorosis (CVC), a disease that is responsible for economic losses in Brazilian agriculture. The most well-known mechanism of pathogenicity for this bacterial pathogen is xylem vessel occlusion, which results from bacterial movement and the formation of biofilms. The molecular mechanisms underlying the virulence caused by biofilm formation are unknown. Here, we provide evidence showing that virulence-associated protein D in X. fastidiosa (Xf-VapD) is a thermostable protein with ribonuclease activity. Moreover, protein expression analyses in two X. fastidiosa strains, including virulent (Xf9a5c) and nonpathogenic (XfJ1a12) strains, showed that Xf-VapD was expressed during all phases of development in both strains and that increased expression was observed in Xf9a5c during biofilm growth. This study is an important step toward characterizing and improving our understanding of the biological significance of Xf-VapD and its potential functions in the CVC pathosystem. PMID:26694028

  11. Population structure of the bacterial pathogen Xylella fastidiosa among street trees in Washington D.C.

    PubMed

    Harris, Jordan Lee; Balci, Yilmaz

    2015-01-01

    Bacterial leaf scorch, associated with the bacterial pathogen Xylella fastidiosa, is a widely established and problematic disease of landscape ornamentals in Washington D.C. A multi-locus sequence typing analysis was performed using 10 housekeeping loci for X. fastidiosa strains in order to better understand the epidemiology of leaf scorch disease in this municipal environment. Samples were collected from 7 different tree species located throughout the District of Columbia, consisting of 101 samples of symptomatic and asymptomatic foliage from 84 different trees. Five strains of the bacteria were identified. Consistent with prior data, these strains were host specific, with only one strain associated with members of the red oak family, one strain associated with American elm, one strain associated with American sycamore, and two strains associated with mulberry. Strains found for asymptomatic foliage were the same as strains from the symptomatic foliage on individual trees. Cross transmission of the strains was not observed at sites with multiple species of infected trees within an approx. 25 m radius of one another. X. fastidiosa strain specificity observed for each genus of tree suggests a highly specialized host-pathogen relationship. PMID:25815838

  12. VapD in Xylella fastidiosa Is a Thermostable Protein with Ribonuclease Activity.

    PubMed

    Mendes, Juliano S; Santiago, André da S; Toledo, Marcelo A S; Rosselli-Murai, Luciana K; Favaro, Marianna T P; Santos, Clelton A; Horta, Maria Augusta C; Crucello, Aline; Beloti, Lilian L; Romero, Fabian; Tasic, Ljubica; de Souza, Alessandra A; de Souza, Anete P

    2015-01-01

    Xylella fastidiosa strain 9a5c is a gram-negative phytopathogen that is the causal agent of citrus variegated chlorosis (CVC), a disease that is responsible for economic losses in Brazilian agriculture. The most well-known mechanism of pathogenicity for this bacterial pathogen is xylem vessel occlusion, which results from bacterial movement and the formation of biofilms. The molecular mechanisms underlying the virulence caused by biofilm formation are unknown. Here, we provide evidence showing that virulence-associated protein D in X. fastidiosa (Xf-VapD) is a thermostable protein with ribonuclease activity. Moreover, protein expression analyses in two X. fastidiosa strains, including virulent (Xf9a5c) and nonpathogenic (XfJ1a12) strains, showed that Xf-VapD was expressed during all phases of development in both strains and that increased expression was observed in Xf9a5c during biofilm growth. This study is an important step toward characterizing and improving our understanding of the biological significance of Xf-VapD and its potential functions in the CVC pathosystem. PMID:26694028

  13. Strain variability in the DNA immigration control region (ICR) of Xylella fastidiosa.

    PubMed

    Picchi, Simone Cristina; Vilas-Boas, Laurival Antonio; Ceresini, Paulo César; de Macedo Lemos, Eliana Gertrudes; Lemos, Manoel Victor Franco

    2006-04-01

    The genome of the bacterium Xylella fastidiosa contains four ORFs (XF2721, XF2725, XF2739 and XF0295) related to the restriction modification type I system, ordinarily named R-M. This system belongs to the DNA immigration control region (ICR). Each ORF is related to different operon structures, which are homologues among themselves and with subunit Hsd R from the endonuclease coding genes. In addition, these ORFs are highly homologous to genes in Pseudomonas aeruginosa, Methylococcus capsulatus str. Bath, Legionella pneumophila, Helicobacter pylori, Xanthomonas oryzae pv. Oryzae and Silicibacter pomeroyi, as well as to genes from X. fastidiosa strains that infect grapevine, almond and oleander plants. This study was carried out on R-M ORFs from forty-three X. fastidiosa strains isolated from citrus, coffee, grapevine, periwinkle, almond and plum trees, in order to assess the genetic diversity of these loci through PCR-RFLP. PCR-RFLP analysis of the four ORFs related to the R-M system from these strains enabled the detection of haplotypes for these loci. When the haplotypes were defined, wide genetic diversity and a large range of similar strains originating from different hosts were observed. This analysis also provided information indicating differences in population genetic structures, which led to detection of different levels of gene transfer among the groups of strains. PMID:16125907

  14. The bacterial pathogen Xylella fastidiosa affects the leaf ionome of plant hosts during infection.

    PubMed

    De La Fuente, Leonardo; Parker, Jennifer K; Oliver, Jonathan E; Granger, Shea; Brannen, Phillip M; van Santen, Edzard; Cobine, Paul A

    2013-01-01

    Xylella fastidiosa is a plant pathogenic bacterium that lives inside the host xylem vessels, where it forms biofilm believed to be responsible for disrupting the passage of water and nutrients. Here, Nicotiana tabacum was infected with X. fastidiosa, and the spatial and temporal changes in the whole-leaf ionome (i.e. the mineral and trace element composition) were measured as the host plant transitioned from healthy to diseased physiological status. The elemental composition of leaves was used as an indicator of the physiological changes in the host at a specific time and relative position during plant development. Bacterial infection was found to cause significant increases in concentrations of calcium prior to the appearance of symptoms and decreases in concentrations of phosphorous after symptoms appeared. Field-collected leaves from multiple varieties of grape, blueberry, and pecan plants grown in different locations over a four-year period in the Southeastern US showed the same alterations in Ca and P. This descriptive ionomics approach characterizes the existence of a mineral element-based response to X. fastidiosa using a model system suitable for further manipulation to uncover additional details of the role of mineral elements during plant-pathogen interactions. This is the first report on the dynamics of changes in the ionome of the host plant throughout the process of infection by a pathogen. PMID:23667547

  15. A new paradigm for vector inoculation of Xylella fastidiosa: Direct evidence of egestion and salivation supports that sharpshooters can be “flying syringes”

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Despite nearly 70 years of research, the inoculation mechanism of Xylella fastidiosa by its sharpshooter vectors remains unproven. X. fastidiosa is unique among insect-transmitted plant pathogens because it is propagative but non-circulative, adhering to and multiplying on the cuticular lining of th...

  16. Hyper-variation of Tandem Repeats at the PD0218 (pspB) locus of Xylella fastidiosa Almond Leaf Scorch and Grape Pierce’s Disease Strains

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Xylella fastidiosa is a Gram negative plant pathogenic bacterium that causes almond leaf scorch disease (ALSD) and grape Pierce’s disease (PD) in California. A-genotype strains cause ALSD only. G-genotype strains cause both PD and ALSD. Little is known about how X. fastidiosa interacts with the envi...

  17. Seasonal Behavior of Xylella fastidiosa Causing Almond Leaf Scorch Disease Under Field Conditions and Improved Detection of the Bacteria by Means of Array-PCR

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Almond leaf scorch disease (ALSD) caused by Xylella fastidiosa is potentially a serious threat to the almond industry in San Joaquin Valley of California. Knowledge of X. fastidiosa behavior in the plant host under field conditions is important for disease control and this issue is being addressed i...

  18. Visualization of Twitching Motility and Characterization of the Role of the PilG in Xylella fastidiosa.

    PubMed

    Shi, Xiangyang; Lin, Hong

    2016-01-01

    Xylella fastidiosa is a Gram-negative non-flagellated bacterium that causes a number of economically important diseases of plants. The twitching motility provides X. fastidiosa a means for long-distance intra-plant movement and colonization, contributing toward pathogenicity in X. fastidiosa. The twitching motility of X. fastidiosa is operated by type IV pili. Type IV pili of Xylella fastidiosa are regulated by pilG, a chemotaxis regulator in Pil-Chp operon encoding proteins that are involved with signal transduction pathways. To elucidate the roles of pilG in the twitching motility of X. fastidiosa, a pilG-deficient mutant XfΔpilG and its complementary strain XfΔpilG-C containing native pilG were developed. A microfluidic chambers integrated with a time-lapse image recording system was used to observe twitching motility in XfΔpilG, XfΔpilG-C and its wild type strain. Using this recording system, it permits long-term spatial and temporal observations of aggregation, migration of individual cells and populations of bacteria via twitching motility. X. fastidiosa wild type and complementary XfΔpilG-C strain showed typical twitching motility characteristics directly observed in the microfluidic flow chambers, whereas mutant XfΔpliG exhibited the twitching deficient phenotype. This study demonstrates that pilG contributes to the twitching motility of X. fastidiosa. The microfluidic flow chamber is used as a means for observing twitching motility. PMID:27166660

  19. Transmission efficiency of Xylella fastidiosa by sharpshooters (Hemiptera: Cicadellidae) in coffee and citrus.

    PubMed

    Marucci, Rosangela C; Lopes, João R S; Cavichioli, Rodney R

    2008-08-01

    Xylella fastidiosa (Wells, Raju, Hung, Weisburg, Mandelco-Paul, and Brenner) is a bacterial pathogen transmitted by several sharpshooters in two tribes of Cicadellinae (Proconiini and Cicadellini). Here, we compared the transmission efficiency of X. fastidiosa in coffee (Coffea arabica L.) and citrus [Citrus sinensis (L.) Osbeck] by Cicadellini [Bucephalogonia xanthophis (Berg) and Dilobopterus costalimai Young] and Proconiini [Homalodisca ignorata Melichar and Oncometopia facialis (Signoret)] sharpshooters that occur in both crops. At different seasons, healthy adults of each species were submitted to a 48-h acquisition access period on citrus or coffee source plants infected with X. fastidiosa isolates that cause Citrus variegated chlorosis (CVC) and Coffee leaf scorch (CLS), respectively, and then confined on healthy seedlings of the corresponding host plant for a 48-h inoculation access period. No significant effect of inoculation season was observed when comparing infection rates of citrus or coffee plants inoculated by vectors at different times of the year. In citrus, the transmission rate by single insects was significantly higher for H. ignorata (30%) in relation to B. xanthophis (5%) and O. facialis (1.1%), but there was no difference among vector species in coffee, whose transmission rates ranged from 1.2 to 7.2%. Comparing host plants, H. ignorata was more effective in transmitting X. fastidiosa to citrus (30%) in relation to coffee (2.2%), whereas the other vectors transmitted the bacterium to both hosts with similar efficiencies. Despite these variations, vector efficiency in coffee and citrus is lower than that reported in other hosts. PMID:18767717

  20. Xylella fastidiosa outer membrane vesicles modulate plant colonization by blocking attachment to surfaces.

    PubMed

    Ionescu, Michael; Zaini, Paulo A; Baccari, Clelia; Tran, Sophia; da Silva, Aline M; Lindow, Steven E

    2014-09-16

    Outer membrane vesicles (OMVs) of Gram-negative bacteria have been studied intensively in recent years, primarily in their role in delivering virulence factors and antigens during pathogenesis. However, the near ubiquity of their production suggests that they may play other roles, such as responding to envelope stress or trafficking various cargoes to prevent dilution or degradation by other bacterial species. Here we show that OMVs produced by Xylella fastidiosa, a xylem-colonizing plant pathogenic bacterium, block its interaction with various surfaces such as the walls of xylem vessels in host plants. The release of OMVs was suppressed by the diffusible signal factor-dependent quorum-sensing system, and a X. fastidiosa ΔrpfF mutant in which quorum signaling was disrupted was both much more virulent to plants and less adhesive to glass and plant surfaces than the WT strain. The higher virulence of the ΔrpfF mutant was associated with fivefold higher numbers of OMVs recovered from xylem sap of infected plants. The frequency of attachment of X. fastidiosa to xylem vessels was 20-fold lower in the presence of OMVs than in their absence. OMV production thus is a strategy used by X. fastidiosa cells to adjust attachment to surfaces in its transition from adhesive cells capable of insect transmission to an "exploratory" lifestyle for systemic spread within the plant host which would be hindered by attachment. OMV production may contribute to the movement of other bacteria in porous environments by similarly reducing their contact with environmental constituents. PMID:25197068

  1. Seasonal Fluctuations of Sap-Feeding Insect Species Infected by Xylella fastidiosa in Apulian Olive Groves of Southern Italy.

    PubMed

    Ben Moussa, Issam Eddine; Mazzoni, Valerio; Valentini, Franco; Yaseen, Thaer; Lorusso, Donato; Speranza, Stefano; Digiaro, Michele; Varvaro, Leonardo; Krugner, Rodrigo; D'Onghia, Anna Maria

    2016-08-01

    A study on seasonal abundance of Auchenorrhyncha species and their infectivity by Xylella fastidiosa in the Apulia region of Italy was conducted to identify ideal periods for monitoring and adoption of potential control measures against insect vectors. Adult populations of Auchenorrhyncha species were monitored monthly over a 2-yr period from five olive groves. A total of 15 species were captured, identified, and tested for presence of X. fastidiosa by polymerase chain reaction (PCR). For three species, Philaenus spumarius L., Neophilaenus campestris (Fallèn), and Euscelis lineolatus Brullé, positive reactions to X. fastidiosa were obtained, on average, in 16.3, 15.9 and 18.4% of adult insects, respectively. Philaneous spumarius was the dominant species (39.8% of total Auchenorrhyncha captured) with the highest adult abundance in summer months. Adult P. spumarius and N. campestris were first detected between March and May in both years, and all insects tested during these periods (year 1: n = 42, year 2: n = 132) gave negative reactions to X. fastidiosa by PCR. Similarly, first adults of E. lineolatus that appeared from October to November (year 1: n = 20, year 2: n = 15) tested negative for presence of X. fastidiosa Given the lack of transstadial and transovarial transmission of X. fastidiosa and considering that P. spumarius is univoltine, control measures against nymphal stages of P. spumarius should be investigated as means of population suppression to reduce spread of X. fastidiosa in olive groves. PMID:27401111

  2. Intersubspecific recombination in Xylella fastidiosa Strains native to the United States: infection of novel hosts associated with an unsuccessful invasion.

    PubMed

    Nunney, Leonard; Hopkins, Donald L; Morano, Lisa D; Russell, Stephanie E; Stouthamer, Richard

    2014-02-01

    The bacterial pathogen Xylella fastidiosa infects xylem and causes disease in many plant species in the Americas. Different subspecies of this bacterium and different genotypes within subspecies infect different plant hosts, but the genetics of host adaptation are unknown. Here we examined the hypothesis that the introduction of novel genetic variation via intersubspecific homologous recombination (IHR) facilitates host shifts. We investigated IHR in 33 X. fastidiosa subsp. multiplex isolates previously identified as recombinant based on 8 loci (7 multilocus sequence typing [MLST] loci plus 1 locus). We found significant evidence of introgression from X. fastidiosa subsp. fastidiosa in 4 of the loci and, using published data, evidence of IHR in 6 of 9 additional loci. Our data showed that IHR regions in 2 of the 4 loci were inconsistent (12 mismatches) with X. fastidiosa subsp. fastidiosa alleles found in the United States but consistent with alleles from Central America. The other two loci were consistent with alleles from both regions. We propose that the recombinant forms all originated via genomewide recombination of one X. fastidiosa subsp. multiplex ancestor with one X. fastidiosa subsp. fastidiosa donor from Central America that was introduced into the United States but subsequently disappeared. Using all of the available data, 5 plant hosts of the recombinant types were identified, 3 of which also supported non-IHR X. fastidiosa subsp. multiplex, but 2 were unique to recombinant types from blueberry (7 isolates from Georgia, 3 from Florida); and blackberry (1 each from Florida and North Carolina), strongly supporting the hypothesis that IHR facilitated a host shift to blueberry and possibly blackberry. PMID:24296499

  3. Intersubspecific Recombination in Xylella fastidiosa Strains Native to the United States: Infection of Novel Hosts Associated with an Unsuccessful Invasion

    PubMed Central

    Hopkins, Donald L.; Morano, Lisa D.; Russell, Stephanie E.; Stouthamer, Richard

    2014-01-01

    The bacterial pathogen Xylella fastidiosa infects xylem and causes disease in many plant species in the Americas. Different subspecies of this bacterium and different genotypes within subspecies infect different plant hosts, but the genetics of host adaptation are unknown. Here we examined the hypothesis that the introduction of novel genetic variation via intersubspecific homologous recombination (IHR) facilitates host shifts. We investigated IHR in 33 X. fastidiosa subsp. multiplex isolates previously identified as recombinant based on 8 loci (7 multilocus sequence typing [MLST] loci plus 1 locus). We found significant evidence of introgression from X. fastidiosa subsp. fastidiosa in 4 of the loci and, using published data, evidence of IHR in 6 of 9 additional loci. Our data showed that IHR regions in 2 of the 4 loci were inconsistent (12 mismatches) with X. fastidiosa subsp. fastidiosa alleles found in the United States but consistent with alleles from Central America. The other two loci were consistent with alleles from both regions. We propose that the recombinant forms all originated via genomewide recombination of one X. fastidiosa subsp. multiplex ancestor with one X. fastidiosa subsp. fastidiosa donor from Central America that was introduced into the United States but subsequently disappeared. Using all of the available data, 5 plant hosts of the recombinant types were identified, 3 of which also supported non-IHR X. fastidiosa subsp. multiplex, but 2 were unique to recombinant types from blueberry (7 isolates from Georgia, 3 from Florida); and blackberry (1 each from Florida and North Carolina), strongly supporting the hypothesis that IHR facilitated a host shift to blueberry and possibly blackberry. PMID:24296499

  4. Nanofilms of hyaluronan/chitosan assembled layer-by-layer: An antibacterial surface for Xylella fastidiosa.

    PubMed

    Hernández-Montelongo, Jacobo; Nascimento, Vicente F; Murillo, Duber; Taketa, Thiago B; Sahoo, Prasana; de Souza, Alessandra A; Beppu, Marisa M; Cotta, Monica A

    2016-01-20

    In this work, nanofilms of hyaluronan/chitosan (HA/CHI) assembled layer by layer were synthesized; their application as a potential antimicrobial material was demonstrated for the phytopathogen Xylella fastidiosa, a gram-negative bacterium, here used as a model. For the synthesis, the influence of pH and ionic strength of these natural polymer stem-solutions on final characteristics of the HA/CHI nanofilms was studied in detail. The antibacterial effect was evaluated using widefield fluorescence microscopy. These results were correlated with the chemical properties of the nanofilms, studied by FTIR and Raman spectroscopy, as well as with their morphology and surface properties characterized using SEM and AFM. The present findings can be extended to design and optimize HA/CHI nanofilms with enhanced antimicrobial behavior for other type of phytopathogenic gram-negative bacteria species, such as Xanthomonas citri, Xanthomas campestri and Ralstonia solanacearum. PMID:26572322

  5. Spatiotemporal distribution of different extracellular polymeric substances and filamentation mediate Xylella fastidiosa adhesion and biofilm formation

    PubMed Central

    Janissen, Richard; Murillo, Duber M.; Niza, Barbara; Sahoo, Prasana K.; Nobrega, Marcelo M.; Cesar, Carlos L.; Temperini, Marcia L. A.; Carvalho, Hernandes F.; de Souza, Alessandra A.; Cotta, Monica A.

    2015-01-01

    Microorganism pathogenicity strongly relies on the generation of multicellular assemblies, called biofilms. Understanding their organization can unveil vulnerabilities leading to potential treatments; spatially and temporally-resolved comprehensive experimental characterization can provide new details of biofilm formation, and possibly new targets for disease control. Here, biofilm formation of economically important phytopathogen Xylella fastidiosa was analyzed at single-cell resolution using nanometer-resolution spectro-microscopy techniques, addressing the role of different types of extracellular polymeric substances (EPS) at each stage of the entire bacterial life cycle. Single cell adhesion is caused by unspecific electrostatic interactions through proteins at the cell polar region, where EPS accumulation is required for more firmly-attached, irreversibly adhered cells. Subsequently, bacteria form clusters, which are embedded in secreted loosely-bound EPS, and bridged by up to ten-fold elongated cells that form the biofilm framework. During biofilm maturation, soluble EPS forms a filamentous matrix that facilitates cell adhesion and provides mechanical support, while the biofilm keeps anchored by few cells. This floating architecture maximizes nutrient distribution while allowing detachment upon larger shear stresses; it thus complies with biological requirements of the bacteria life cycle. Using new approaches, our findings provide insights regarding different aspects of the adhesion process of X. fastidiosa and biofilm formation. PMID:25891045

  6. The exopolysaccharide of Xylella fastidiosa is essential for biofilm formation, plant virulence, and vector transmission.

    PubMed

    Killiny, N; Martinez, R Hernandez; Dumenyo, C Korsi; Cooksey, D A; Almeida, R P P

    2013-09-01

    Exopolysaccharides (EPS) synthesized by plant-pathogenic bacteria are generally essential for virulence. The role of EPS produced by the vector-transmitted bacterium Xylella fastidiosa was investigated by knocking out two genes implicated in the EPS biosynthesis, gumD and gumH. Mutant strains were affected in growth characteristics in vitro, including adhesion to surfaces and biofilm formation. In addition, different assays were used to demonstrate that the mutant strains produced significantly less EPS compared with the wild type. Furthermore, gas chromatography-mass spectrometry showed that both mutant strains did not produce oligosaccharides. Biologically, the mutants were deficient in movement within plants, resulting in an avirulent phenotype. Additionally, mutant strains were affected in transmission by insects: they were very poorly transmitted by and retained within vectors. The gene expression profile indicated upregulation of genes implicated in cell-to-cell signaling and adhesins while downregulation in genes was required for within-plant movement in EPS-deficient strains. These results suggest an essential role for EPS in X. fastidiosa interactions with both plants and insects. PMID:23678891

  7. Spatiotemporal distribution of different extracellular polymeric substances and filamentation mediate Xylella fastidiosa adhesion and biofilm formation.

    PubMed

    Janissen, Richard; Murillo, Duber M; Niza, Barbara; Sahoo, Prasana K; Nobrega, Marcelo M; Cesar, Carlos L; Temperini, Marcia L A; Carvalho, Hernandes F; de Souza, Alessandra A; Cotta, Monica A

    2015-01-01

    Microorganism pathogenicity strongly relies on the generation of multicellular assemblies, called biofilms. Understanding their organization can unveil vulnerabilities leading to potential treatments; spatially and temporally-resolved comprehensive experimental characterization can provide new details of biofilm formation, and possibly new targets for disease control. Here, biofilm formation of economically important phytopathogen Xylella fastidiosa was analyzed at single-cell resolution using nanometer-resolution spectro-microscopy techniques, addressing the role of different types of extracellular polymeric substances (EPS) at each stage of the entire bacterial life cycle. Single cell adhesion is caused by unspecific electrostatic interactions through proteins at the cell polar region, where EPS accumulation is required for more firmly-attached, irreversibly adhered cells. Subsequently, bacteria form clusters, which are embedded in secreted loosely-bound EPS, and bridged by up to ten-fold elongated cells that form the biofilm framework. During biofilm maturation, soluble EPS forms a filamentous matrix that facilitates cell adhesion and provides mechanical support, while the biofilm keeps anchored by few cells. This floating architecture maximizes nutrient distribution while allowing detachment upon larger shear stresses; it thus complies with biological requirements of the bacteria life cycle. Using new approaches, our findings provide insights regarding different aspects of the adhesion process of X. fastidiosa and biofilm formation. PMID:25891045

  8. Expression of Xylella fastidiosa RpfF in citrus disrupts signaling in Xanthomonas citri subsp. citri and thereby its virulence.

    PubMed

    Caserta, R; Picchi, S C; Takita, M A; Tomaz, J P; Pereira, W E L; Machado, M A; Ionescu, M; Lindow, S; De Souza, A A

    2014-11-01

    Xylella fastidiosa and Xanthomonas citri subsp. citri, that cause citrus variegated chlorosis (CVC) and citrus canker diseases, respectively, utilize diffusible signal factor (DSF) for quorum sensing. DSF, produced by RpfF, are similar fatty acids in both organisms, although a different set of genes is regulated by DSF in each species. Because of this similarity, Xylella fastidiosa DSF might be recognized and affect the biology of Xanthomonas citri. Therefore, transgenic Citrus sinensis and Carrizo citrange plants overexpressing the Xylella fastidiosa rpfF were inoculated with Xanthomonas citri and changes in symptoms of citrus canker were observed. X. citri biofilms formed only at wound sites on transgenic leaves and were thicker; however, bacteria were unable to break through the tissue and form pustules elsewhere. Although abundant growth of X. citri occurred at wound sites on inoculated transgenic leaves, little growth was observed on unwounded tissue. Genes in the DFS-responsive core in X. citri were downregulated in bacteria isolated from transgenic leaves. DSF-dependent expression of engA was suppressed in cells exposed to xylem sap from transgenic plants. Thus, altered symptom development appears to be due to reduced expression of virulence genes because of the presence of antagonists of DSF signaling in X. citri in rpfF-expressing plants. PMID:25099341

  9. Analyses of Xylella whole genome sequences and proposal of Xylella taiwanensis sp. nov.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Xylella fastidiosa is a Gram negative, xylem limited and nutritionally fastidious plant pathogenic bacterium that cause disease in many economically important plants. A single species, fastidiosa, with three subspecies (fastidiosa, multiplex, and pauca) have been described. Most Xylella strains were...

  10. O antigen modulates insect vector acquisition of the bacterial plant pathogen Xylella fastidiosa.

    PubMed

    Rapicavoli, Jeannette N; Kinsinger, Nichola; Perring, Thomas M; Backus, Elaine A; Shugart, Holly J; Walker, Sharon; Roper, M Caroline

    2015-12-01

    Hemipteran insect vectors transmit the majority of plant pathogens. Acquisition of pathogenic bacteria by these piercing/sucking insects requires intimate associations between the bacterial cells and insect surfaces. Lipopolysaccharide (LPS) is the predominant macromolecule displayed on the cell surface of Gram-negative bacteria and thus mediates bacterial interactions with the environment and potential hosts. We hypothesized that bacterial cell surface properties mediated by LPS would be important in modulating vector-pathogen interactions required for acquisition of the bacterial plant pathogen Xylella fastidiosa, the causative agent of Pierce's disease of grapevines. Utilizing a mutant that produces truncated O antigen (the terminal portion of the LPS molecule), we present results that link this LPS structural alteration to a significant decrease in the attachment of X. fastidiosa to blue-green sharpshooter foreguts. Scanning electron microscopy confirmed that this defect in initial attachment compromised subsequent biofilm formation within vector foreguts, thus impairing pathogen acquisition. We also establish a relationship between O antigen truncation and significant changes in the physiochemical properties of the cell, which in turn affect the dynamics of X. fastidiosa adhesion to the vector foregut. Lastly, we couple measurements of the physiochemical properties of the cell with hydrodynamic fluid shear rates to produce a Comsol model that predicts primary areas of bacterial colonization within blue-green sharpshooter foreguts, and we present experimental data that support the model. These results demonstrate that, in addition to reported protein adhesin-ligand interactions, O antigen is crucial for vector-pathogen interactions, specifically in the acquisition of this destructive agricultural pathogen. PMID:26386068

  11. O Antigen Modulates Insect Vector Acquisition of the Bacterial Plant Pathogen Xylella fastidiosa

    PubMed Central

    Rapicavoli, Jeannette N.; Kinsinger, Nichola; Perring, Thomas M.; Backus, Elaine A.; Shugart, Holly J.; Walker, Sharon

    2015-01-01

    Hemipteran insect vectors transmit the majority of plant pathogens. Acquisition of pathogenic bacteria by these piercing/sucking insects requires intimate associations between the bacterial cells and insect surfaces. Lipopolysaccharide (LPS) is the predominant macromolecule displayed on the cell surface of Gram-negative bacteria and thus mediates bacterial interactions with the environment and potential hosts. We hypothesized that bacterial cell surface properties mediated by LPS would be important in modulating vector-pathogen interactions required for acquisition of the bacterial plant pathogen Xylella fastidiosa, the causative agent of Pierce's disease of grapevines. Utilizing a mutant that produces truncated O antigen (the terminal portion of the LPS molecule), we present results that link this LPS structural alteration to a significant decrease in the attachment of X. fastidiosa to blue-green sharpshooter foreguts. Scanning electron microscopy confirmed that this defect in initial attachment compromised subsequent biofilm formation within vector foreguts, thus impairing pathogen acquisition. We also establish a relationship between O antigen truncation and significant changes in the physiochemical properties of the cell, which in turn affect the dynamics of X. fastidiosa adhesion to the vector foregut. Lastly, we couple measurements of the physiochemical properties of the cell with hydrodynamic fluid shear rates to produce a Comsol model that predicts primary areas of bacterial colonization within blue-green sharpshooter foreguts, and we present experimental data that support the model. These results demonstrate that, in addition to reported protein adhesin-ligand interactions, O antigen is crucial for vector-pathogen interactions, specifically in the acquisition of this destructive agricultural pathogen. PMID:26386068

  12. Production of Xylella fastidiosa diffusible signal factor in transgenic grape causes pathogen confusion and reduction in severity of Pierce's disease.

    PubMed

    Lindow, Steven; Newman, Karyn; Chatterjee, Subhadeep; Baccari, Clelia; Lavarone, Anthony T; Ionescu, Michael

    2014-03-01

    The rpfF gene from Xylella fastidiosa, encoding the synthase for diffusible signal factor (DSF), was expressed in 'Freedom' grape to reduce the pathogen's growth and mobility within the plant. Symptoms in such plants were restricted to near the point of inoculation and incidence of disease was two- to fivefold lower than in the parental line. Both the longitudinal and lateral movement of X. fastidiosa in the xylem was also much lower. DSF was detected in both leaves and xylem sap of RpfF-expressing plants using biological sensors, and both 2-Z-tetradecenoic acid, previously identified as a component of X. fastidiosa DSF, and cis-11-methyl-2-dodecenoic acid were detected in xylem sap using electrospray ionization mass spectrometry. A higher proportion of X. fastidiosa cells adhered to xylem vessels of the RpfF-expressing line than parental 'Freedom' plants, reflecting a higher adhesiveness of the pathogen in the presence of DSF. Disease incidence in RpfF-expressing plants in field trials in which plants were either mechanically inoculated with X. fastidiosa or subjected to natural inoculation by sharpshooter vectors was two- to fourfold lower in than that of the parental line. The number of symptomatic leaves on infected shoots was reduced proportionally more than the incidence of infection, reflecting a decreased ability of X. fastidiosa to move within DSF-producing plants. PMID:24499029

  13. Population Structure of Xylella fastidiosa Associated with Almond Leaf Scorch Disease in the San Joaquin Valley of California.

    PubMed

    Lin, Hong; Islam, Md Sajedul; Cabrera-La Rosa, Juan C; Civerolo, Edwin L; Groves, Russell L

    2015-06-01

    Xylella fastidiosa causes disease in many commercial crops, including almond leaf scorch (ALS) disease in susceptible almond (Prunus dulcis). In this study, genetic diversity and population structure of X. fastidiosa associated with ALS disease were evaluated. Isolates obtained from two almond orchards in Fresno and Kern County in the San Joaquin Valley of California were analyzed for two successive years. Multilocus simple-sequence repeat (SSR) analysis revealed two major genetic clusters that were associated with two host cultivars, 'Sonora' and 'Nonpareil', respectively, regardless of the year of study or location of the orchard. These relationships suggest that host cultivar selection and adaptation are major driving forces shaping ALS X. fastidiosa population structure in the San Joaquin Valley. This finding will provide insight into understanding pathogen adaptation and host selection in the context of ALS disease dynamics. PMID:25807309

  14. Ionome changes in Xylella fastidiosa-infected Nicotiana tabacum correlate with virulence and discriminate between subspecies of bacterial isolates.

    PubMed

    Oliver, J E; Sefick, S A; Parker, J K; Arnold, T; Cobine, P A; De La Fuente, L

    2014-10-01

    Characterization of ionomes has been used to uncover the basis of nutrient utilization and environmental adaptation of plants. Here, ionomic profiles were used to understand the phenotypic response of a plant to infection by genetically diverse isolates of Xylella fastidiosa, a gram-negative, xylem-limited bacterial plant pathogen. In this study, X. fastidiosa isolates were used to infect a common model host (Nicotiana tabacum 'SR1'), and leaf and sap concentrations of eleven elements together with plant colonization and symptoms were assessed. Multivariate statistical analysis revealed that changes in the ionome were significantly correlated with symptom severity and bacterial populations in host petioles. Moreover, plant ionome modification by infection could be used to differentiate the X. fastidiosa subspecies with which the plant was infected. This report establishes host ionome modification as a phenotypic response to infection. PMID:24983508

  15. Calcium-Enhanced Twitching Motility in Xylella fastidiosa Is Linked to a Single PilY1 Homolog.

    PubMed

    Cruz, Luisa F; Parker, Jennifer K; Cobine, Paul A; De La Fuente, Leonardo

    2014-12-01

    The plant-pathogenic bacterium Xylella fastidiosa is restricted to the xylem vessel environment, where mineral nutrients are transported through the plant host; therefore, changes in the concentrations of these elements likely impact the growth and virulence of this bacterium. Twitching motility, dependent on type IV pili (TFP), is required for movement against the transpiration stream that results in basipetal colonization. We previously demonstrated that calcium (Ca) increases the motility of X. fastidiosa, although the mechanism was unknown. PilY1 is a TFP structural protein recently shown to bind Ca and to regulate twitching and adhesion in bacterial pathogens of humans. Sequence analysis identified three pilY1 homologs in X. fastidiosa (PD0023, PD0502, and PD1611), one of which (PD1611) contains a Ca-binding motif. Separate deletions of PD0023 and PD1611 resulted in mutants that still showed twitching motility and were not impaired in attachment or biofilm formation. However, the response of increased twitching at higher Ca concentrations was lost in the pilY1-1611 mutant. Ca does not modulate the expression of any of the X. fastidiosa PilY1 homologs, although it increases the expression of the retraction ATPase pilT during active movement. The evidence presented here suggests functional differences between the PilY1 homologs, which may provide X. fastidiosa with an adaptive advantage in environments with high Ca concentrations, such as xylem sap. PMID:25217013

  16. Initial biochemical and functional characterization of a 5'-nucleotidase from Xylella fastidiosa related to the human cytosolic 5'-nucleotidase I.

    PubMed

    Santos, Clelton A; Saraiva, Antonio M; Toledo, Marcelo A S; Beloti, Lilian L; Crucello, Aline; Favaro, Marianna T P; Horta, Maria A C; Santiago, André S; Mendes, Juliano S; Souza, Alessandra A; Souza, Anete P

    2013-01-01

    The 5'-nucleotidases constitute a ubiquitous family of enzymes that catalyze either the hydrolysis or the transfer of esterified phosphate at the 5' position of nucleoside monophosphates. These enzymes are responsible for the regulation of nucleotide and nucleoside levels in the cell and can interfere with the phosphorylation-dependent activation of nucleoside analogs used in therapies targeting solid tumors and viral infections. In the present study, we report the initial biochemical and functional characterization of a 5'-nucleotidase from Xylella fastidiosa that is related to the human cytosolic 5'-nucleotidase I. X. fastidiosa is a plant pathogenic bacterium that is responsible for numerous economically important crop diseases. Biochemical assays confirmed the phosphatase activity of the recombinant purified enzyme and revealed metal ion dependence for full enzyme activity. In addition, we investigated the involvement of Xf5'-Nt in the formation of X. fastidiosa biofilms, which are structures that occlude the xylem vessels of susceptible plants and are strictly associated with bacterial pathogenesis. Using polyclonal antibodies against Xf5'-Nt, we observed an overexpression of Xf5'-Nt during the initial phases of X. fastidiosa biofilm formation that was not observed during X. fastidiosa planktonic growth. Our results demonstrate that the de/phosphorylation network catalyzed by 5'-nucleotidases may play an important role in bacterial biofilm formation, thereby contributing novel insights into bacterial nucleotide metabolism and pathogenicity. PMID:23474016

  17. Calcium-Enhanced Twitching Motility in Xylella fastidiosa Is Linked to a Single PilY1 Homolog

    PubMed Central

    Cruz, Luisa F.; Parker, Jennifer K.; Cobine, Paul A.

    2014-01-01

    The plant-pathogenic bacterium Xylella fastidiosa is restricted to the xylem vessel environment, where mineral nutrients are transported through the plant host; therefore, changes in the concentrations of these elements likely impact the growth and virulence of this bacterium. Twitching motility, dependent on type IV pili (TFP), is required for movement against the transpiration stream that results in basipetal colonization. We previously demonstrated that calcium (Ca) increases the motility of X. fastidiosa, although the mechanism was unknown. PilY1 is a TFP structural protein recently shown to bind Ca and to regulate twitching and adhesion in bacterial pathogens of humans. Sequence analysis identified three pilY1 homologs in X. fastidiosa (PD0023, PD0502, and PD1611), one of which (PD1611) contains a Ca-binding motif. Separate deletions of PD0023 and PD1611 resulted in mutants that still showed twitching motility and were not impaired in attachment or biofilm formation. However, the response of increased twitching at higher Ca concentrations was lost in the pilY1-1611 mutant. Ca does not modulate the expression of any of the X. fastidiosa PilY1 homologs, although it increases the expression of the retraction ATPase pilT during active movement. The evidence presented here suggests functional differences between the PilY1 homologs, which may provide X. fastidiosa with an adaptive advantage in environments with high Ca concentrations, such as xylem sap. PMID:25217013

  18. A Temperature-Independent Cold-Shock Protein Homolog Acts as a Virulence Factor in Xylella fastidiosa.

    PubMed

    Burbank, Lindsey P; Stenger, Drake C

    2016-05-01

    Xylella fastidiosa, causal agent of Pierce's disease (PD) of grapevine, is a fastidious organism that requires very specific conditions for replication and plant colonization. Cold temperatures reduce growth and survival of X. fastidiosa both in vitro and in planta. However, little is known regarding physiological responses of X. fastidiosa to temperature changes. Cold-shock proteins (CSP), a family of nucleic acid-binding proteins, act as chaperones facilitating translation at low temperatures. Bacterial genomes often encode multiple CSP, some of which are strongly induced following exposure to cold. Additionally, CSP contribute to the general stress response through mRNA stabilization and posttranscriptional regulation. A putative CSP homolog (Csp1) with RNA-binding activity was identified in X. fastidiosa Stag's Leap. The csp1 gene lacked the long 5' untranslated region characteristic of cold-inducible genes and was expressed in a temperature-independent manner. As compared with the wild type, a deletion mutant of csp1 (∆csp1) had decreased survival rates following cold exposure and salt stress in vitro. The deletion mutant also was significantly less virulent in grapevine, as compared with the wild type, in the absence of cold stress. These results suggest an important function of X. fastidiosa Csp1 in response to cellular stress and during plant colonization. PMID:26808446

  19. Structural and biochemical characterization of peroxiredoxin Qbeta from Xylella fastidiosa: catalytic mechanism and high reactivity.

    PubMed

    Horta, Bruno Brasil; de Oliveira, Marcos Antonio; Discola, Karen Fulan; Cussiol, José Renato Rosa; Netto, Luis Eduardo Soares

    2010-05-21

    The phytopathogenic bacterium Xylella fastidiosa is the etiological agent of various plant diseases. To survive under oxidative stress imposed by the host, microorganisms express antioxidant proteins, including cysteine-based peroxidases named peroxiredoxins. This work is a comprehensive analysis of the catalysis performed by PrxQ from X. fastidiosa (XfPrxQ) that belongs to a peroxiredoxin class still poorly characterized and previously considered as moderately reactive toward hydroperoxides. Contrary to these assumptions, our competitive kinetics studies have shown that the second-order rate constants of the peroxidase reactions of XfPrxQ with hydrogen peroxide and peroxynitrite are in the order of 10(7) and 10(6) M(-1) S(-1), respectively, which are as fast as the most efficient peroxidases. The XfPrxQ disulfides were only slightly reducible by dithiothreitol; therefore, the identification of a thioredoxin system as the probable biological reductant of XfPrxQ was a relevant finding. We also showed by site-specific mutagenesis and mass spectrometry that an intramolecular disulfide bond between Cys-47 and Cys-83 is generated during the catalytic cycle. Furthermore, we elucidated the crystal structure of XfPrxQ C47S in which Ser-47 and Cys-83 lie approximately 12.3 A apart. Therefore, significant conformational changes are required for disulfide bond formation. In fact, circular dichroism data indicated that there was a significant redox-dependent unfolding of alpha-helices, which is probably triggered by the peroxidatic cysteine oxidation. Finally, we proposed a model that takes data from this work as well data as from the literature into account. PMID:20335172

  20. Calcium Increases Xylella fastidiosa Surface Attachment, Biofilm Formation, and Twitching Motility

    PubMed Central

    Cruz, Luisa F.; Cobine, Paul A.

    2012-01-01

    Xylella fastidiosa is a plant-pathogenic bacterium that forms biofilms inside xylem vessels, a process thought to be influenced by the chemical composition of xylem sap. In this work, the effect of calcium on the production of X. fastidiosa biofilm and movement was analyzed under in vitro conditions. After a dose-response study with 96-well plates using eight metals, the strongest increase of biofilm formation was observed when medium was supplemented with at least 1.0 mM CaCl2. The removal of Ca by extracellular (EGTA, 1.5 mM) and intracellular [1,2-bis(o-aminophenoxy)ethane-N,N,N′,N′-tetraacetic acid acetoxymethyl ester (BAPTA/AM), 75 μM] chelators reduced biofilm formation without compromising planktonic growth. The concentration of Ca influenced the force of adhesion to the substrate, biofilm thickness, cell-to-cell aggregation, and twitching motility, as shown by assays with microfluidic chambers and other assays. The effect of Ca on attachment was lost when cells were treated with tetracycline, suggesting that Ca has a metabolic or regulatory role in cell adhesion. A double mutant (fimA pilO) lacking type I and type IV pili did not improve biofilm formation or attachment when Ca was added to the medium, while single mutants of type I (fimA) or type IV (pilB) pili formed more biofilm under conditions of higher Ca concentrations. The concentration of Ca in the medium did not significantly influence the levels of exopolysaccharide produced. Our findings indicate that the role of Ca in biofilm formation may be related to the initial surface and cell-to-cell attachment and colonization stages of biofilm establishment, which rely on critical functions by fimbrial structures. PMID:22194297

  1. Microarray Analyses of Xylella fastidiosa Provide Evidence of Coordinated Transcription Control of Laterally Transferred Elements

    PubMed Central

    Nunes, Luiz R.; Rosato, Yoko B.; Muto, Nair H.; Yanai, Giane M.; da Silva, Vivian S.; Leite, Daniela B.; Gonçalves, Edmilson R.; de Souza, Alessandra A.; Coletta-Filho, Helvécio D.; Machado, Marcos A.; Lopes, Silvio A.; de Oliveira, Regina Costa

    2003-01-01

    Genetically distinct strains of the plant bacterium Xylella fastidiosa (Xf) are responsible for a variety of plant diseases, accounting for severe economic damage throughout the world. Using as a reference the genome of Xf 9a5c strain, associated with citrus variegated chlorosis (CVC), we developed a microarray-based comparison involving 12 Xf isolates, providing a thorough assessment of the variation in genomic composition across the group. Our results demonstrate that Xf displays one of the largest flexible gene pools characterized to date, with several horizontally acquired elements, such as prophages, plasmids, and genomic islands (GIs), which contribute up to 18% of the final genome. Transcriptome analysis of bacteria grown under different conditions shows that most of these elements are transcriptionally active, and their expression can be influenced in a coordinated manner by environmental stimuli. Finally, evaluation of the genetic composition of these laterally transferred elements identified differences that may help to explain the adaptability of Xf strains to infect such a wide range of plant species. [Supplemental material is available online at www.genome.org.] PMID:12670998

  2. Grape Cultivar and Sap Culture Conditions Affect the Development of Xylella fastidiosa Phenotypes Associated with Pierce's Disease.

    PubMed

    Hao, Lingyun; Zaini, Paulo A; Hoch, Harvey C; Burr, Thomas J; Mowery, Patricia

    2016-01-01

    Xylella fastidiosa is a xylem-limited bacterium in plant hosts and causes Pierce's disease (PD) of grapevines, which differ in susceptibility according to the Vitis species (spp.). In this work we compared X. fastidiosa biofilm formation and population dynamics when cultured in xylem saps from PD-susceptible and -resistant Vitis spp. under different conditions. Behaviors in a closed-culture system were compared to those in different sap-renewal cultures that would more closely mimic the physicochemical environment encountered in planta. Significant differences in biofilm formation and growth in saps from PD-susceptible and -resistant spp. were only observed using sap renewal culture. Compared to saps from susceptible V. vinifera, those from PD-resistant V. aestivalis supported lower titers of X. fastidiosa and less biofilm and V. champinii suppressed both growth and biofilm formation, behaviors which are correlated with disease susceptibility. Furthermore, in microfluidic chambers X. fastidiosa formed thick mature biofilm with three-dimensional (3-D) structures, such as pillars and mounds, in saps from all susceptible spp. In contrast, only small aggregates of various shapes were formed in saps from four out of five of the resistant spp.; sap from the resistant spp. V. mustangensis was an exception in that it also supported thick lawns of biofilm but not the above described 3-D structures typically seen in a mature biofilm from the susceptible saps. Our findings provide not only critical technical information for future bioassays, but also suggest further understanding of PD susceptibility. PMID:27508296

  3. Grape Cultivar and Sap Culture Conditions Affect the Development of Xylella fastidiosa Phenotypes Associated with Pierce's Disease

    PubMed Central

    Hoch, Harvey C.; Burr, Thomas J.; Mowery, Patricia

    2016-01-01

    Xylella fastidiosa is a xylem-limited bacterium in plant hosts and causes Pierce’s disease (PD) of grapevines, which differ in susceptibility according to the Vitis species (spp.). In this work we compared X. fastidiosa biofilm formation and population dynamics when cultured in xylem saps from PD-susceptible and -resistant Vitis spp. under different conditions. Behaviors in a closed-culture system were compared to those in different sap-renewal cultures that would more closely mimic the physicochemical environment encountered in planta. Significant differences in biofilm formation and growth in saps from PD-susceptible and -resistant spp. were only observed using sap renewal culture. Compared to saps from susceptible V. vinifera, those from PD-resistant V. aestivalis supported lower titers of X. fastidiosa and less biofilm and V. champinii suppressed both growth and biofilm formation, behaviors which are correlated with disease susceptibility. Furthermore, in microfluidic chambers X. fastidiosa formed thick mature biofilm with three-dimensional (3-D) structures, such as pillars and mounds, in saps from all susceptible spp. In contrast, only small aggregates of various shapes were formed in saps from four out of five of the resistant spp.; sap from the resistant spp. V. mustangensis was an exception in that it also supported thick lawns of biofilm but not the above described 3-D structures typically seen in a mature biofilm from the susceptible saps. Our findings provide not only critical technical information for future bioassays, but also suggest further understanding of PD susceptibility. PMID:27508296

  4. Structural and Biochemical Characterization of Xylella fastidiosa DsbA Family Members: New insightsinto the Enzyme-Substrate Interaction

    SciTech Connect

    Rinaldi, F.; Meza, A; Gulmarges, B

    2009-01-01

    Disulfide oxidoreductase DsbA catalyzes disulfide bond formation in proteins secreted to the periplasm and has been related to the folding process of virulence factors in many organisms. It is among the most oxidizing of the thioredoxin-like proteins, and DsbA redox power is understood in terms of the electrostatic interactions involving the active site motif CPHC. The plant pathogen Xylella fastidiosa has two chromosomal genes encoding two oxidoreductases belonging to the DsbA family, and in one of them, the canonical motif CPHC is replaced by CPAC. Biochemical assays showed that both X. fastidiosa homologues have similar redox properties and the determination of the crystal structure of XfDsbA revealed substitutions in the active site of X. fastidiosa enzymes, which are proposed to compensate for the lack of the conserved histidine in XfDsbA2. In addition, electron density maps showed a ligand bound to the XfDsbA active site, allowing the characterization of the enzyme interaction with an 8-mer peptide. Finally, surface analysis of XfDsbA and XfDsbA2 suggests that X. fastidiosa enzymes may have different substrate specificities.

  5. Response of Xylella fastidiosa to zinc: decreased culturability, increased exopolysaccharide production, and formation of resilient biofilms under flow conditions.

    PubMed

    Navarrete, Fernando; De La Fuente, Leonardo

    2014-02-01

    The bacterial plant pathogen Xylella fastidiosa produces biofilm that accumulates in the host xylem vessels, affecting disease development in various crops and bacterial acquisition by insect vectors. Biofilms are sensitive to the chemical composition of the environment, and mineral elements being transported in the xylem are of special interest for this pathosystem. Here, X. fastidiosa liquid cultures were supplemented with zinc and compared with nonamended cultures to determine the effects of Zn on growth, biofilm, and exopolysaccharide (EPS) production under batch and flow culture conditions. The results show that Zn reduces growth and biofilm production under both conditions. However, in microfluidic chambers under liquid flow and with constant bacterial supplementation (closer to conditions inside the host), a dramatic increase in biofilm aggregates was seen in the Zn-amended medium. Biofilms formed under these conditions were strongly attached to surfaces and were not removed by medium flow. This phenomenon was correlated with increased EPS production in stationary-phase cells grown under high Zn concentrations. Zn did not cause greater adhesion to surfaces by individual cells. Additionally, viability analyses suggest that X. fastidiosa may be able to enter the viable but nonculturable state in vitro, and Zn can hasten the onset of this state. Together, these findings suggest that Zn can act as a stress factor with pleiotropic effects on X. fastidiosa and indicate that, although Zn could be used as a bactericide treatment, it could trigger the undesired effect of stronger biofilm formation upon reinoculation events. PMID:24271184

  6. Plasmid Vectors for Xylella fastidiosa Utilizing a Toxin-Antitoxin System for Stability in the Absence of Antibiotic Selection.

    PubMed

    Burbank, Lindsey P; Stenger, Drake C

    2016-08-01

    The phytopathogen Xylella fastidiosa causes disease in a variety of important crop and landscape plants. Functional genetic studies have led to a broader understanding of virulence mechanisms used by this pathogen in the grapevine host. Plasmid shuttle vectors are important tools in studies of bacterial genetics but there are only a limited number of plasmid vectors available that replicate in X. fastidiosa, and even fewer that are retained without antibiotic selection. Two plasmids are described here that show stable replication in X. fastidiosa and are effective for gene complementation both in vitro and in planta. Plasmid maintenance is facilitated by incorporation of the PemI/PemK plasmid addiction system, consisting of PemK, an endoribonuclease toxin, and its cognate antitoxin, PemI. Vector pXf20pemIK utilizes a native X. fastidiosa replication origin as well as a high-copy-number pUC origin for propagation in Escherichia coli cloning strains. Broad-host-range vector pBBR5pemIK is a medium- to low-copy-number plasmid based on the pBBR1 backbone. Both plasmids are maintained for extended periods of time in the absence of antibiotic selection, as well as up to 14 weeks in grapevine, without affecting bacterial fitness. These plasmids present an alternative to traditional complementation and expression vectors which rely on antibiotic selection for plasmid retention. PMID:27088393

  7. Characterization of Growth and Virulence-related Genes Expression of Xylella fastidiosa Affected by Grape Xylem Sap and Cell-Wall Constituents

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Pierce’s disease (PD) of grapevines is caused by the xylem-limited bacterium Xylella fastidiosa (Xf). Most Vitis vinifera-based cultivars are susceptible to PD. However, grape species such as V. arizonica, V. shuttleworthii, V. simpsonii, V. smalliana, and Muscadinia rotundifolia are resistant to PD...

  8. Seasonal Behavior of Xylella fastidiosa Causing Almond Leaf Scorch Under Field Conditions and Detection of the Bacteria by Means of Array-PCR

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Diseases caused by Xylella fastidiosa have re-emerged as a serious threat to several economically important crops, such as grape and almond, in the San Joaquin Valley of California. Knowledge of bacterial behavior in plant hosts under field condition is important for disease control. This research c...

  9. Mulberry strains of Xylella fastidiosa contain a 25 kilobase pair plasmid with extensive sequence identity to a plasmid from Verminephrobacter eiseniae

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A 25 kbp plasmid was present in each of four Californian strains of Xylella fastidiosa from mulberry affected with leaf scorch disease. Fragments of each plasmid were cloned into E. coli, sequenced, and assembled into circular contigs of 25,105 bp (pXF-RIV11 and pXF-RIV16) or 24,372 bp (pXF-RIV19 an...

  10. Xylella fastidiosa isolates from mulberry harbor a 25 kilobase pair plasmid with extensive sequence identity to a plasmid from Verminephrobacter eiseniae

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A 25 kbp plasmid was present in each of four Californian isolates of Xylella fastidiosa from mulberry affected with leaf scorch disease. Fragments of each plasmid were cloned into E. coli, sequenced, and assembled into circular contigs of 25,105 bp (pXF-RIV11 and pXF-RIV16) or 24,372 bp (pXF-RIV19 a...

  11. Plant water stress effects on stylet probing behaviors of Homalodisca vitripennis (Hemiptera: Cicadellidae) associated with acquisition and inoculation of the bacterium Xylella fastidiosa

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The glassy-winged sharpshooter, Homalodisca vitripennis, is a xylem fluid-ingesting leafhopper that transmits Xylella fastidiosa, the causal agent of several plant diseases in the Americas. While the role of plant water stress on the population density and dispersal of H. vitripennis has been studie...

  12. Population dynamics of insect vectors of Xylella fastidiosa in almond nurseries in the San Joaquin Valley of California: An assessment of plant vulnerability to almond leaf scorch disease.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Almond leaf scorch (ALS) is caused by the xylem-limited bacterium Xylella fastidiosa, which also infects ~145 other plant species in California. The pathogen is transmitted by xylem-fluid feeding insects including sharpshooters and spittlebugs. Within almond orchards tree-to-tree spread of the pat...

  13. Lipopolysaccharide modulates the vector-pathogen interface of the xylem-limited phytopathogen, Xylella fastidiosa, the causal agent of Pierce’s disease of grapevine

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Xylella fastidiosa Wells et al. is a gram-negative, insect-transmitted bacterium that causes a lethal disease of grapevine called Pierce’s disease. Lipopolysaccharide (LPS) is the most dominant macromolecule displayed on the cell surface of gram-negative bacteria. Bacterial interactions with the env...

  14. Plasmids of Xylella fastidiosa Mulberry-Infecting Strains Share Extensive Sequence Identity and Gene Complement with pVEIS01 From the Earthworm Symbiont Verminephrobacter Eiseniae

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A ~25 kbp plasmid was present in each of four Californian strains of Xylella fastidiosa from mulberry affected with leaf scorch disease. Fragments of each plasmid were cloned into E. coli, sequenced, and assembled into circular contigs of 25,105 bp (pXF-RIV11 and pXF-RIV16) or 24,372 bp (pXF-RIV19 a...

  15. Development and systematic validation of qPCR assays for rapid and reliable differentiation of Xylella fastidiosa strains causing citrus variegated chlorosis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The xylem-limited, Gram-negative, fastidious plant bacterium Xylella fastidiosa is the causal agent of citrus variegated chlorosis (CVC), a destructive disease affecting approximately half of the citrus plantations in the State of São Paulo, Brazil. The disease was recently found in Central America...

  16. Muscle movements that control inoculation of Xylella fastidiosa are revealed by head X-rays of feeding glassy-winged sharpshooters

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The mechanism of inoculation of the Pierce’s disease bacterium, Xylella fastidiosa (Xf), by vectors such as the glassy-winged sharpshooter (GWSS) is still unknown, despite nearly 70 years of study. Research in support of the egestion-salivation hypothesis for Xf inoculation is presented. Two impor...

  17. Whole genome evaluation of tandem repeat polymorphisms between two pathogenically similar strains of Xylella fastidiosa isolated from almond and grape in California

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Whole genome tandem repeat polymorphisms were evaluated between two closely related Xylella fastidiosa strains, M23 and Temecula1, both cause almond leaf scorch disease (ALSD) and grape Pierce’s disease (PD) in California. Strain M23 was isolated from almond and the genome was sequenced in this stu...

  18. Time course of colonization of the glassy-winged sharpshooter precibarium by Xylella fastidiosa provides evidence for two types of egestion during inoculation

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Introduction of the glassy-winged sharpshooter (GWSS) into California has caused an increase in incidence of Pierce’s Disease in grape-growing regions. Although host plant resistance to the causative bacterium, Xylella fastidiosa, and/or its GWSS vector is being sought, research is hampered by lack...

  19. Genetic variation of Xylella fastidiosa associated with grape vines in two major viticulture regions in the United Sates: California and Texas

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Xylella fastidiosa (Xf) causes Pierce’s disease in grapevine. Here, genetic diversity and population structure of grape strains of Xf collected from two important grape growing regions in the United States, California and Texas, is presented. Multilocus simple sequence repeat (SSR) markers revealed ...

  20. Effects of Grape Xylem Sap and Cell Wall Constituents on In Vitro Growth, Biofilm Formation, and Cellular Aggregation of Xylella fastidiosa

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Pierce’s Disease (PD) of grapevines is caused by the xylem-limited bacterium Xylella fastidiosa (Xf). Most Vitis vinifera-based cultivars are susceptible to PD; however, related grape species V. arizonica, V. shuttleworthii, V. simpsonii, V. smalliana, and Muscadinia rotundifolia are PD resistant. ...

  1. Effects of Grape Xylem Sap and Cell-Wall Constituents on In Vitro Growth, Biofilm Formation and Cellular Aggregation of Xylella fastidiosa

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Purified cell-wall constituents or grape xylem sap added to media affected in vitro growth, biofilm formation, cell aggregation and gene expression of Xylella fastidiosa. Media containing xylem sap from Pierce’s disease (PD)-susceptible plants provided better support for bacterial growth and biofil...

  2. In vitro Study of the Effects of Grape Xylem Saps and Cell-Wall Constituents on Growth and Virulence-Related Gene Expression of Xylella fastidiosa

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Pierce’s disease (PD) of grapevines is caused by the xylem-inhabiting bacterium Xylella fastidiosa. The widely cultivated Vitis vinifera-based grape cultivars are susceptible to PD. However, grape species V. aestivalis, V. arizonica, V. shuttleworthii, V. simpsonii, V. smalliana, and Muscadinia rotu...

  3. Phenology of Xylella fastidiosa and Draeculocephala minerva in California almond nurseries: an assessment of plant vulnerability to almond leaf scorch disease

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Management of a plant disease requires knowledge of all possible infection pathways. Almond leaf scorch disease (ALSD) is caused by the xylem-limited bacterium Xylella fastidiosa, which is transmitted by several species of leafhoppers. The objectives of this research were to elucidate the fate of b...

  4. Phenology of Xylella fastidiosa and its vector around California almond nurseries: An assessment of plant vulnerability to almond leaf scorch disease

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Management of almond leaf scorch disease requires knowledge of all possible infection pathways. The disease is caused by the xylem-limited bacterium Xylella fastidiosa, which is transmitted by several species of leafhoppers. The objectives of this research were to elucidate the fate of bacteria in p...

  5. Wintercuring of Prunus dulcis cv ‘Butte,’ P. webbii and their interspecific hybrid in response to Xylella fastidiosa infections

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Clonal replicates of Prunus dulcis cv ‘Butte,’ P. webbii and their interspecific hybrid P 63-61 were inoculated with Xylella fastidiosa strain M23 and evaluated for Almond Leaf Scorch Disease and subsequent wintercuring of infections during three growing seasons. Initial inoculations established gr...

  6. Xylella fastidiosa is associated with reduced yield and altered fruit quality in a rabbiteye blueberry orchard in Louisiana but does not appear to spread rapidly

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Xylella fastidiosa is a bacterial pathogen that causes diseases such as Pierce’s disease of grape, bacterial leaf scorch of shade trees, and citrus variegated chlorosis. Work by researchers in Georgia and Florida has shown that it is the cause of bacterial leaf scorch of Southern highbush blueberrie...

  7. Host selection and adaptation are major driving forces shaping ALS Xylella fastidiosa population structure in the San Joaquin Valley of California

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Xylella fastidiosa (Xf) causes disease in many commercial crops including almond leaf scorch (ALS) disease in susceptible almond (Prunus dulcis). In this study, genetic diversity and population structure of Xf associated with ALS disease were evaluated. Strains from two almond production sites in th...

  8. Development of single chain variable fragment (scFv) antibodies against Xylella fastidiosa subsp. pauca by phage display.

    PubMed

    Yuan, Qing; Jordan, Ramon; Brlansky, Ronald H; Istomina, Olga; Hartung, John

    2015-10-01

    Xylella fastidiosa is a member of the gamma proteobacteria. It is fastidious, insect-vectored and xylem-limited and causes a variety of diseases, some severe, on a wide range of economically important perennial crops, including grape and citrus. Antibody based detection assays are commercially available for X. fastidiosa, and are effective at the species, but not at the subspecies level. We have made a library of scFv antibody fragments directed against X. fastidiosa subsp. pauca strain 9a5c (citrus) by using phage display technology. Antibody gene repertoires were PCR-amplified using 23 primers for the heavy chain variable region (V(H)) and 21 primers for the light chain variable region (V(L)). The V(H) and V(L) were joined by overlap extension PCR, and then the genes of the scFv library were ligated into the phage vector pKM19. The library contained 1.2×10(7) independent clones with full-length scFv inserts. In each of 3cycles of affinity-selection with 9a5c, about 1.0×10(12) phage were used for panning with 4.1×10(6), 7.1×10(6), 2.1×10(7) phage recovered after the first, second and third cycles, respectively. Sixty-six percent of clones from the final library bound X. fastidiosa 9a5c in an ELISA. Some of these scFv antibodies recognized strain 9a5c and did not recognize X. fastidiosa strains that cause Pierce's disease of grapevine. PMID:26232710

  9. Factors affecting the initial adhesion and retention of the plant pathogen Xylella fastidiosa in the foregut of an insect vector.

    PubMed

    Killiny, Nabil; Almeida, Rodrigo P P

    2014-01-01

    Vector transmission of bacterial plant pathogens involves three steps: pathogen acquisition from an infected host, retention within the vector, and inoculation of cells into susceptible tissue of an uninfected plant. In this study, a combination of plant and artificial diet systems were used to determine the importance of several genes on the initial adhesion and retention of the bacterium Xylella fastidiosa to an efficient insect vector. Mutant strains included fimbrial (fimA and pilB) and afimbrial (hxfA and hxfB) adhesins and three loci involved in regulatory systems (rpfF, rpfC, and cgsA). Transmission assays with variable retention time indicated that HxfA and HxfB were primarily important for early adhesion to vectors, while FimA was necessary for both adhesion and retention. The long pilus protein PilB was not deficient in initial adhesion but may be important for retention. Genes upregulated under the control of rpfF are important for both initial adhesion and retention, as transmission rates of this mutant strain were initially low and decreased over time, while disruption of rpfC and cgsA yielded trends similar to that shown by the wild-type control. Because induction of an X. fastidiosa transmissible state requires pectin, a series of experiments were used to test the roles of a polygalacturonase (pglA) and the pectin and galacturonic acid carbohydrates on the transmission of X. fastidiosa. Results show that galacturonic acid, or PglA activity breaking pectin into its major subunit (galacturonic acid), is required for X. fastidiosa vector transmission using an artificial diet system. This study shows that early adhesion and retention of X. fastidiosa are mediated by different factors. It also illustrates that the interpretation of results of vector transmission experiments, in the context of vector-pathogen interaction studies, is highly dependent on experimental design. PMID:24185853

  10. Seasonal Abundance and Natural Inoculativity of Insect Vectors of Xylella fastidiosa in Oklahoma Tree Nurseries and Vineyards.

    PubMed

    Overall, Lisa M; Rebek, Eric J

    2015-12-01

    Xylella fastidiosa is the causative agent of diseases of perennial plants including peach, plum, elm, oak, pecan, and grape. This bacterial pathogen is transmitted by xylem-feeding insects. In recent years, Pierce's disease of grape has been detected in 10 counties in central and northeastern Oklahoma, prompting further investigation of the disease epidemiology in this state. We surveyed vineyards and tree nurseries in Oklahoma for potential insect vectors to determine species composition, infectivity, and natural inoculativity of commonly captured insect vectors. Yellow sticky cards were used to sample insect fauna at each location. Insects were removed from sticky cards and screened for X. fastidiosa using immunocapture-PCR to determine their infectivity. A second objective was to test the natural inoculativity of insect vectors that are found in vineyards. Graphocephala versuta (Say), Graphocephala coccinea (Forster), Paraulacizes irrorata (F.), Oncometopia orbona (F.), Cuerna costalis (F.), and Entylia carinata Germar were collected from vineyards and taken back to the lab to determine their natural inoculativity. Immunocapture-PCR was used to test plant and insect samples for presence of X. fastidiosa. The three most frequently captured species from vineyards and tree nurseries were G. versuta, Clastoptera xanthocephala Germar, and O. orbona. Of those insects screened for X. fastidiosa, 2.4% tested positive for the bacterium. Field-collected G. versuta were inoculative to both ragweed and alfalfa. Following a 7-d inoculation access period, a higher percentage of alfalfa became infected than ragweed. Results from this study provide insight into the epidemiology of X. fastidiosa in Oklahoma. PMID:26331482

  11. Citrus Functional Genomics and Molecular Modeling in Relation to Citrus sinensis (Sweet Orange) Infection with Xylella fastidiosa (Citrus Variegated Chlorosis).

    PubMed

    Dwivedi, Upendra N; Tiwari, Sameeksha; Prasanna, Pragya; Awasthi, Manika; Singh, Swati; Pandey, Veda P

    2016-08-01

    Citrus are among the economically most important fruit tree crops in the world. Citrus variegated chlorosis (CVC), caused by Xylella fastidiosa infection, is a serious disease limiting citrus production at a global scale. With availability of citrus genomic resources, it is now possible to compare citrus expressed sequence tag (EST) data sets and identify single-nucleotide polymorphisms (SNPs) within and among different citrus cultivars that can be exploited for citrus resistance to infections, citrus breeding, among others. We report here, for the first time, SNPs in the EST data sets of X. fastidiosa-infected Citrus sinensis (sweet orange) and their functional annotation that revealed the involvement of eight C. sinensis candidate genes in CVC pathogenesis. Among these genes were xyloglucan endotransglycosylase, myo-inositol-1-phosphate synthase, and peroxidase were found to be involved in plant cell wall metabolism. These have been further investigated by molecular modeling for their role in CVC infection and defense. Molecular docking analyses of the wild and the mutant (SNP containing) types of the selected three enzymes with their respective substrates revealed a significant decrease in the binding affinity of substrates for the mutant enzymes, thus suggesting a decrease in the catalytic efficiency of these enzymes during infection, thereby facilitating a favorable condition for infection by the pathogen. These findings offer novel agrigenomics insights in developing future molecular targets and strategies for citrus fruit cultivation in ways that are resistant to X. fastidiosa infection, and by extension, with greater harvesting efficiency and economic value. PMID:27447755

  12. Calcium transcriptionally regulates the biofilm machinery of Xylella fastidiosa to promote continued biofilm development in batch cultures.

    PubMed

    Parker, Jennifer K; Chen, Hongyu; McCarty, Sara E; Liu, Lawrence Y; De La Fuente, Leonardo

    2016-05-01

    The functions of calcium (Ca) in bacteria are less characterized than in eukaryotes, where its role has been studied extensively. The plant-pathogenic bacterium Xylella fastidiosa has several virulence features that are enhanced by increased Ca concentrations, including biofilm formation. However, the specific mechanisms driving modulation of this feature are unclear. Characterization of biofilm formation over time showed that 4 mM Ca supplementation produced denser biofilms that were still developing at 96 h, while biofilm in non-supplemented media had reached the dispersal stage by 72 h. To identify changes in global gene expression in X. fastidiosa grown in supplemental Ca, RNA-Seq of batch culture biofilm cells was conducted at three 24-h time intervals. Results indicate that a variety of genes are differentially expressed in response to Ca, including genes related to attachment, motility, exopolysaccharide synthesis, biofilm formation, peptidoglycan synthesis, regulatory functions, iron homeostasis, and phages. Collectively, results demonstrate that Ca supplementation induces a transcriptional response that promotes continued biofilm development, while biofilm cells in nonsupplemented media are driven towards dispersion of cells from the biofilm structure. These results have important implications for disease progression in planta, where xylem sap is the source of Ca and other nutrients for X. fastidiosa. PMID:26913481

  13. Zinc Detoxification Is Required for Full Virulence and Modification of the Host Leaf Ionome by Xylella fastidiosa.

    PubMed

    Navarrete, Fernando; De La Fuente, Leonardo

    2015-04-01

    Zinc (Zn) is an essential element for all forms of life because it is a structural or catalytic cofactor of many proteins, but it can have toxic effects at high concentrations; thus, microorganisms must tightly regulate its levels. Here, we evaluated the role of Zn homeostasis proteins in the virulence of the xylem-limited bacterium Xylella fastidiosa, causal agent of Pierce's disease of grapevine, among other diseases. Two mutants of X. fastidiosa 'Temecula' affected in genes which regulate Zn homeostasis (zur) and Zn detoxification (czcD) were constructed. Both knockouts showed increased sensitivity to Zn at physiologically relevant concentrations and increased intracellular accumulation of this metal compared with the wild type. Increased Zn sensitivity was correlated with decreased growth in grapevine xylem sap, reduced twitching motility, and downregulation of exopolysaccharide biosynthetic genes. Tobacco plants inoculated with either knockout mutant showed reduced foliar symptoms and a much reduced (czcD) or absent (zur) modification of the leaf ionome (i.e., the mineral nutrient and trace element composition), as well as reduced bacterial populations. The results show that detoxification of Zn is crucial for the virulence of X. fastidiosa and verifies our previous findings that modification of the host leaf ionome correlates with bacterial virulence. PMID:25561271

  14. Insights into the Activity and Substrate Binding of Xylella fastidiosa Polygalacturonase by Modification of a Unique QMK Amino Acid Motif Using Protein Chimeras.

    PubMed

    Warren, Jeremy G; Lincoln, James E; Kirkpatrick, Bruce C

    2015-01-01

    Polygalacturonases (EC 3.2.1.15) catalyze the random hydrolysis of 1, 4-alpha-D-galactosiduronic linkages in pectate and other galacturonans. Xylella fastidiosa possesses a single polygalacturonase gene, pglA (PD1485), and X. fastidiosa mutants deficient in the production of polygalacturonase are non-pathogenic and show a compromised ability to systemically infect grapevines. These results suggested that grapevines expressing sufficient amounts of an inhibitor of X. fastidiosa polygalacturonase might be protected from disease. Previous work in our laboratory and others have tried without success to produce soluble active X. fastidiosa polygalacturonase for use in inhibition assays. In this study, we created two enzymatically active X. fastidiosa / A. vitis polygalacturonase chimeras, AX1A and AX2A to explore the functionality of X. fastidiosa polygalacturonase in vitro. The AX1A chimera was constructed to specifically test if recombinant chimeric protein, produced in Escherichia coli, is soluble and if the X. fastidiosa polygalacturonase catalytic amino acids are able to hydrolyze polygalacturonic acid. The AX2A chimera was constructed to evaluate the ability of a unique QMK motif of X. fastidiosa polygalacturonase, most polygalacturonases have a R(I/L)K motif, to bind to and allow the hydrolysis of polygalacturonic acid. Furthermore, the AX2A chimera was also used to explore what effect modification of the QMK motif of X. fastidiosa polygalacturonase to a conserved RIK motif has on enzymatic activity. These experiments showed that both the AX1A and AX2A polygalacturonase chimeras were soluble and able to hydrolyze the polygalacturonic acid substrate. Additionally, the modification of the QMK motif to the conserved RIK motif eliminated hydrolytic activity, suggesting that the QMK motif is important for the activity of X. fastidiosa polygalacturonase. This result suggests X. fastidiosa polygalacturonase may preferentially hydrolyze a different pectic substrate or

  15. Specific Detection and Identification of American Mulberry-Infecting and Italian Olive-Associated Strains of Xylella fastidiosa by Polymerase Chain Reaction.

    PubMed

    Guan, Wei; Shao, Jonathan; Elbeaino, Toufic; Davis, Robert E; Zhao, Tingchang; Huang, Qi

    2015-01-01

    Xylella fastidiosa causes bacterial leaf scorch in many landscape trees including elm, oak, sycamore and mulberry, but methods for specific identification of a particular tree host species-limited strain or differentiation of tree-specific strains are lacking. It is also unknown whether a particular landscape tree-infecting X. fastidiosa strain is capable of infecting multiple landscape tree species in an urban environment. We developed two PCR primers specific for mulberry-infecting strains of X. fastidiosa based on the nucleotide sequence of a unique open reading frame identified only in mulberry-infecting strains among all the North and South American strains of X. fastidiosa sequenced to date. PCR using the primers allowed for detection and identification of mulberry-infecting X. fastidiosa strains in cultures and in samples collected from naturally infected mulberry trees. In addition, no mixed infections with or non-specific detections of the mulberry-infecting strains of X. fastidiosa were found in naturally X. fastidiosa-infected oak, elm and sycamore trees growing in the same region where naturally infected mulberry trees were grown. This genotype-specific PCR assay will be valuable for disease diagnosis, studies of strain-specific infections in insects and plant hosts, and management of diseases caused by X. fastidiosa. Unexpectedly but interestingly, the unique open reading frame conserved in the mulberry-infecting strains in the U. S. was also identified in the recently sequenced olive-associated strain CoDiRO isolated in Italy. When the primer set was tested against naturally infected olive plant samples collected in Italy, it allowed for detection of olive-associated strains of X. fastidiosa in Italy. This PCR assay, therefore, will also be useful for detection and identification of the Italian group of X. fastidiosa strains to aid understanding of the occurrence, evolution and biology of this new group of X. fastidiosa strains. PMID:26061051

  16. Insights into the Activity and Substrate Binding of Xylella fastidiosa Polygalacturonase by Modification of a Unique QMK Amino Acid Motif Using Protein Chimeras

    PubMed Central

    Warren, Jeremy G.; Lincoln, James E.; Kirkpatrick, Bruce C.

    2015-01-01

    Polygalacturonases (EC 3.2.1.15) catalyze the random hydrolysis of 1, 4-alpha-D-galactosiduronic linkages in pectate and other galacturonans. Xylella fastidiosa possesses a single polygalacturonase gene, pglA (PD1485), and X. fastidiosa mutants deficient in the production of polygalacturonase are non-pathogenic and show a compromised ability to systemically infect grapevines. These results suggested that grapevines expressing sufficient amounts of an inhibitor of X. fastidiosa polygalacturonase might be protected from disease. Previous work in our laboratory and others have tried without success to produce soluble active X. fastidiosa polygalacturonase for use in inhibition assays. In this study, we created two enzymatically active X. fastidiosa / A. vitis polygalacturonase chimeras, AX1A and AX2A to explore the functionality of X. fastidiosa polygalacturonase in vitro. The AX1A chimera was constructed to specifically test if recombinant chimeric protein, produced in Escherichia coli, is soluble and if the X. fastidiosa polygalacturonase catalytic amino acids are able to hydrolyze polygalacturonic acid. The AX2A chimera was constructed to evaluate the ability of a unique QMK motif of X. fastidiosa polygalacturonase, most polygalacturonases have a R(I/L)K motif, to bind to and allow the hydrolysis of polygalacturonic acid. Furthermore, the AX2A chimera was also used to explore what effect modification of the QMK motif of X. fastidiosa polygalacturonase to a conserved RIK motif has on enzymatic activity. These experiments showed that both the AX1A and AX2A polygalacturonase chimeras were soluble and able to hydrolyze the polygalacturonic acid substrate. Additionally, the modification of the QMK motif to the conserved RIK motif eliminated hydrolytic activity, suggesting that the QMK motif is important for the activity of X. fastidiosa polygalacturonase. This result suggests X. fastidiosa polygalacturonase may preferentially hydrolyze a different pectic substrate or

  17. Specific Detection and Identification of American Mulberry-Infecting and Italian Olive-Associated Strains of Xylella fastidiosa by Polymerase Chain Reaction

    PubMed Central

    Guan, Wei; Shao, Jonathan; Elbeaino, Toufic; Davis, Robert E.; Zhao, Tingchang; Huang, Qi

    2015-01-01

    Xylella fastidiosa causes bacterial leaf scorch in many landscape trees including elm, oak, sycamore and mulberry, but methods for specific identification of a particular tree host species-limited strain or differentiation of tree-specific strains are lacking. It is also unknown whether a particular landscape tree-infecting X. fastidiosa strain is capable of infecting multiple landscape tree species in an urban environment. We developed two PCR primers specific for mulberry-infecting strains of X. fastidiosa based on the nucleotide sequence of a unique open reading frame identified only in mulberry-infecting strains among all the North and South American strains of X. fastidiosa sequenced to date. PCR using the primers allowed for detection and identification of mulberry-infecting X. fastidiosa strains in cultures and in samples collected from naturally infected mulberry trees. In addition, no mixed infections with or non-specific detections of the mulberry-infecting strains of X. fastidiosa were found in naturally X. fastidiosa-infected oak, elm and sycamore trees growing in the same region where naturally infected mulberry trees were grown. This genotype-specific PCR assay will be valuable for disease diagnosis, studies of strain-specific infections in insects and plant hosts, and management of diseases caused by X. fastidiosa. Unexpectedly but interestingly, the unique open reading frame conserved in the mulberry-infecting strains in the U. S. was also identified in the recently sequenced olive-associated strain CoDiRO isolated in Italy. When the primer set was tested against naturally infected olive plant samples collected in Italy, it allowed for detection of olive-associated strains of X. fastidiosa in Italy. This PCR assay, therefore, will also be useful for detection and identification of the Italian group of X. fastidiosa strains to aid understanding of the occurrence, evolution and biology of this new group of X. fastidiosa strains. PMID:26061051

  18. Transmission ecology and epidemiology of the citrus variegated chlorosis strain of Xylella fastidiosa

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Xylella fastidosa is a vector-borne vascular bacterium that inhabits the xylem vessels of a wide range of host plants. Most of the information on vector-Xylella-plant interactions derives from studies with strains that cause Pierce’s disease of grapevine in the United States. The subspecies fastid...

  19. PemK toxin encoded by the Xylella fastidiosa IncP-1 plasmid pXF-RIV11 is a ribonuclease

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Stable inheritance of the IncP-1 plasmid pXF-RIV11 in Xylella fastidiosa is conferred by the pemI/pemK plasmid addiction system. PemK serves as a toxin inhibiting bacterial growth; PemI is the corresponding antitoxin that blocks activity of PemK toxin by direct binding. Here, PemK toxin and PemI ant...

  20. Diffusible signal factor (DSF) synthase RpfF of Xylella fastidiosa is a multifunction protein also required for response to DSF.

    PubMed

    Ionescu, Michael; Baccari, Clelia; Da Silva, Aline Maria; Garcia, Angelica; Yokota, Kenji; Lindow, Steven E

    2013-12-01

    Xylella fastidiosa, like related Xanthomonas species, employs an Rpf cell-cell communication system consisting of a diffusible signal factor (DSF) synthase, RpfF, and a DSF sensor, RpfC, to coordinate expression of virulence genes. While phenotypes of a ΔrpfF strain in Xanthomonas campestris could be complemented by its own DSF, the DSF produced by X. fastidiosa (XfDSF) did not restore expression of the XfDSF-dependent genes hxfA and hxfB to a ΔrpfF strain of X. fastidiosa, suggesting that RpfF is involved in XfDSF sensing or XfDSF-dependent signaling. To test this conjecture, rpfC and rpfF of X. campestris were replaced by those of X. fastidiosa, and the contribution of each gene to the induction of a X. campestris DSF-dependent gene was assessed. As in X. fastidiosa, XfDSF-dependent signaling required both X. fastidiosa proteins RpfF and RpfC. RpfF repressed RpfC signaling activity, which in turn was derepressed by XfDSF. A mutated X. fastidiosa RpfF protein with two substitutions of glutamate to alanine in its active site was incapable of XfDSF production yet enabled a response to XfDSF, indicating that XfDSF production and the response to XfDSF are two separate functions in which RpfF is involved. This mutant was also hypervirulent to grape, demonstrating the antivirulence effects of XfDSF itself in X. fastidiosa. The Rpf system of X. fastidiosa is thus a novel example of a quorum-sensing signal synthase that is also involved in the response to the signal molecule that it synthesizes. PMID:24056101

  1. Characterization of an oxidative stress response regulator, homologous to Escherichia coli OxyR, from the phytopathogen Xylella fastidiosa.

    PubMed

    Toledo, M A S; Schneider, D R; Azzoni, A R; Favaro, M T P; Pelloso, A C; Santos, C A; Saraiva, A M; Souza, A P

    2011-02-01

    The OxyR oxidative stress transcriptional regulator is a DNA-binding protein that belongs to the LysR-type transcriptional regulators (LTTR) family. It has the ability to sense oxidative species inside the cell and to trigger the cell's response, activating the transcription of genes involved in scavenging oxidative species. In the present study, we have overexpressed, purified and characterized the predicted OxyR homologue (orf xf1273) of the phytopathogen Xylella fastidiosa. This bacterium is the causal agent of citrus variegated chlorosis (CVC) disease caused by the 9a5c strain, resulting in economic and social losses. The secondary structure of the recombinant protein was analyzed by circular dichroism. Gel filtration showed that XfoxyR is a dimer in solution. Gel shift assays indicated that it does bind to its own predicted promoter under in vitro conditions. However, considering our control experiment we cannot state that this interaction occurs in vivo. Functional complementation assays indicated that xfoxyR is able to restore the oxidative stress response in an oxyr knockout Escherichia coli strain. These results show that the predicted orfxf1273 codes for a transcriptional regulator, homologous to E. coli OxyR, involved in the oxidative stress response. This may be important for X. fastidiosa to overcome the defense mechanisms of its host during the infection and colonization processes. PMID:20951212

  2. Characterization of the LysR-type transcriptional regulator YcjZ-like from Xylella fastidiosa overexpressed in Escherichia coli.

    PubMed

    Santiago, André S; Santos, Clelton A; Mendes, Juliano S; Toledo, Marcelo A S; Beloti, Lilian L; Souza, Alessandra A; Souza, Anete P

    2015-09-01

    The Xylella fastidiosa 9a5c strain is a xylem-limited phytopathogen that is the causal agent of citrus variegated chlorosis (CVC). This bacterium is able to form a biofilm and occlude the xylem vessels of susceptible plants, which leads to significant agricultural and economic losses. Biofilms are associated with bacterial pathogenicity because they are very resistant to antibiotics and other metal-based chemicals that are used in agriculture. The X. fastidiosa YcjZ-like (XfYcjZ-like) protein belongs to the LysR-type transcriptional regulator (LTTR) family and is involved in various cellular functions that range from quorum sensing to bacterial survival. In the present study, we report the cloning, expression and purification of XfYcjZ-like, which was overexpressed in Escherichia coli. The secondary folding of the recombinant and purified protein was assessed by circular dichroism, which revealed that XfYcjZ-like contains a typical α/β fold. An initial hydrodynamic characterization showed that XfYcjZ-like is a globular tetramer in solution. In addition, using a polyclonal antibody against XfYcjZ-like, we assessed the expression profile of this protein during the different developmental phases of X. fastidiosa in in vitro cultivated biofilm cells and demonstrated that XfYcjZ-like is upregulated in planktonic cells in response to a copper shock treatment. Finally, the ability of XfYcjZ-like to interact with its own predicted promoter was confirmed in vitro, which is a typical feature of LysR. Taken together, our findings indicated that the XfYcjZ-like protein is involved in both the organization of the architecture and the maturation of the bacterial biofilm and that it is responsive to oxidative stress. PMID:25979465

  3. Large-scale intersubspecific recombination in the plant-pathogenic bacterium Xylella fastidiosa is associated with the host shift to mulberry.

    PubMed

    Nunney, Leonard; Schuenzel, Erin L; Scally, Mark; Bromley, Robin E; Stouthamer, Richard

    2014-05-01

    Homologous recombination plays an important role in the structuring of genetic variation of many bacteria; however, its importance in adaptive evolution is not well established. We investigated the association of intersubspecific homologous recombination (IHR) with the shift to a novel host (mulberry) by the plant-pathogenic bacterium Xylella fastidiosa. Mulberry leaf scorch was identified about 25 years ago in native red mulberry in the eastern United States and has spread to introduced white mulberry in California. Comparing a sequence of 8 genes (4,706 bp) from 21 mulberry-type isolates to published data (352 isolates representing all subspecies), we confirmed previous indications that the mulberry isolates define a group distinct from the 4 subspecies, and we propose naming the taxon X. fastidiosa subsp. morus. The ancestry of its gene sequences was mixed, with 4 derived from X. fastidiosa subsp. fastidiosa (introduced from Central America), 3 from X. fastidiosa subsp. multiplex (considered native to the United States), and 1 chimeric, demonstrating that this group originated by large-scale IHR. The very low within-type genetic variation (0.08% site polymorphism), plus the apparent inability of native X. fastidiosa subsp. multiplex to infect mulberry, suggests that this host shift was achieved after strong selection acted on genetic variants created by IHR. Sequence data indicate that a single ancestral IHR event gave rise not only to X. fastidiosa subsp. morus but also to the X. fastidiosa subsp. multiplex recombinant group which infects several hosts but is the only type naturally infecting blueberry, thus implicating this IHR in the invasion of at least two novel native hosts, mulberry and blueberry. PMID:24610840

  4. Characterization of the Xylella fastidiosa PD1671 gene encoding degenerate c-di-GMP GGDEF/EAL domains, and its role in the development of Pierce's disease.

    PubMed

    Cursino, Luciana; Athinuwat, Dusit; Patel, Kelly R; Galvani, Cheryl D; Zaini, Paulo A; Li, Yaxin; De La Fuente, Leonardo; Hoch, Harvey C; Burr, Thomas J; Mowery, Patricia

    2015-01-01

    Xylella fastidiosa is an important phytopathogenic bacterium that causes many serious plant diseases including Pierce's disease of grapevines. X. fastidiosa is thought to induce disease by colonizing and clogging xylem vessels through the formation of cell aggregates and bacterial biofilms. Here we examine the role in X. fastidiosa virulence of an uncharacterized gene, PD1671, annotated as a two-component response regulator with potential GGDEF and EAL domains. GGDEF domains are found in c-di-GMP diguanylate cyclases while EAL domains are found in phosphodiesterases, and these domains are for c-di-GMP production and turnover, respectively. Functional analysis of the PD1671 gene revealed that it affected multiple X. fastidiosa virulence-related phenotypes. A Tn5 PD1671 mutant had a hypervirulent phenotype in grapevines presumably due to enhanced expression of gum genes leading to increased exopolysaccharide levels that resulted in elevated biofilm formation. Interestingly, the PD1671 mutant also had decreased motility in vitro but did not show a reduced distribution in grapevines following inoculation. Given these responses, the putative PD1671 protein may be a negative regulator of X. fastidiosa virulence. PMID:25811864

  5. Differential gene expression in Xylella fastidiosa 9a5c during co-cultivation with the endophytic bacterium Methylobacterium mesophilicum SR1.6/6.

    PubMed

    Dourado, Manuella Nóbrega; Santos, Daiene Souza; Nunes, Luiz Roberto; Costa de Oliveira, Regina Lúcia Batista da; de Oliveira, Marcus Vinicius; Araújo, Welington Luiz

    2015-12-01

    Xylella fastidiosa, the causal agent of citrus variegated chlorosis (CVC), colonizes plant xylem, reducing sap flow, and inducing internerval chlorosis, leaf size reduction, necrosis, and harder and smaller fruits. This bacterium may be transmitted from plant to plant by sharpshooter insects, including Bucephalogonia xanthopis. The citrus endophytic bacterium Methylobacterium mesophilicum SR1.6/6 colonizes citrus xylem and previous studies showed that this strain is also transferred from plant to plant by B. xanthopis (Insecta), suggesting that this endophytic bacterium may interact with X. fastidiosa in planta and inside the insect vector during co-transmission by the same insect vector. To better understand the X. fastidiosa behavior in the presence of M. mesophilicum, we evaluated the X. fastidiosa transcriptional profile during in vitro interaction with M. mesophilicum SR1.6/6. The results showed that during co-cultivation, X. fastidiosa down-regulated genes related to growth and up-regulated genes related to energy production, stress, transport, and motility, suggesting the existence of a specific adaptive response to the presence of M. mesophilicum in the culture medium. PMID:26218710

  6. Recent Evolutionary Radiation and Host Plant Specialization in the Xylella fastidiosa Subspecies Native to the United States

    PubMed Central

    Vickerman, Danel B.; Bromley, Robin E.; Russell, Stephanie A.; Hartman, John R.; Morano, Lisa D.; Stouthamer, Richard

    2013-01-01

    The bacterial pathogen, Xylella fastidiosa, infects many plant species in the Americas, making it a good model for investigating the genetics of host adaptation. We used multilocus sequence typing (MLST) to identify isolates of the native U.S. subsp. multiplex that were largely unaffected by intersubspecific homologous recombination (IHR) and to investigate how their evolutionary history influences plant host specialization. We identified 110 “non-IHR” isolates, 2 minimally recombinant “intermediate” ones (including the subspecific type), and 31 with extensive IHR. The non-IHR and intermediate isolates defined 23 sequence types (STs) which we used to identify 22 plant hosts (73% trees) characteristic of the subspecies. Except for almond, subsp. multiplex showed no host overlap with the introduced subspecies (subspecies fastidiosa and sandyi). MLST sequences revealed that subsp. multiplex underwent recent radiation (<25% of subspecies age) which included only limited intrasubspecific recombination (ρ/θ = 0.02); only one isolated lineage (ST50 from ash) was older. A total of 20 of the STs grouped into three loose phylogenetic clusters distinguished by nonoverlapping hosts (excepting purple leaf plum): “almond,” “peach,” and “oak” types. These host differences were not geographical, since all three types also occurred in California. ST designation was a good indicator of host specialization. ST09, widespread in the southeastern United States, only infected oak species, and all peach isolates were ST10 (from California, Florida, and Georgia). Only ST23 had a broad host range. Hosts of related genotypes were sometimes related, but often host groupings crossed plant family or even order, suggesting that phylogenetically plastic features of hosts affect bacterial pathogenicity. PMID:23354698

  7. Recent evolutionary radiation and host plant specialization in the Xylella fastidiosa subspecies native to the United States.

    PubMed

    Nunney, Leonard; Vickerman, Danel B; Bromley, Robin E; Russell, Stephanie A; Hartman, John R; Morano, Lisa D; Stouthamer, Richard

    2013-04-01

    The bacterial pathogen, Xylella fastidiosa, infects many plant species in the Americas, making it a good model for investigating the genetics of host adaptation. We used multilocus sequence typing (MLST) to identify isolates of the native U.S. subsp. multiplex that were largely unaffected by intersubspecific homologous recombination (IHR) and to investigate how their evolutionary history influences plant host specialization. We identified 110 "non-IHR" isolates, 2 minimally recombinant "intermediate" ones (including the subspecific type), and 31 with extensive IHR. The non-IHR and intermediate isolates defined 23 sequence types (STs) which we used to identify 22 plant hosts (73% trees) characteristic of the subspecies. Except for almond, subsp. multiplex showed no host overlap with the introduced subspecies (subspecies fastidiosa and sandyi). MLST sequences revealed that subsp. multiplex underwent recent radiation (<25% of subspecies age) which included only limited intrasubspecific recombination (ρ/θ = 0.02); only one isolated lineage (ST50 from ash) was older. A total of 20 of the STs grouped into three loose phylogenetic clusters distinguished by nonoverlapping hosts (excepting purple leaf plum): "almond," "peach," and "oak" types. These host differences were not geographical, since all three types also occurred in California. ST designation was a good indicator of host specialization. ST09, widespread in the southeastern United States, only infected oak species, and all peach isolates were ST10 (from California, Florida, and Georgia). Only ST23 had a broad host range. Hosts of related genotypes were sometimes related, but often host groupings crossed plant family or even order, suggesting that phylogenetically plastic features of hosts affect bacterial pathogenicity. PMID:23354698

  8. Specific detection and identification of mulberry-infecting strains of Xylella fastidiosa by polymerase chain reaction

    Technology Transfer Automated Retrieval System (TEKTRAN)

    X. fastidiosa causes bacterial leaf scorch in many landscape trees including elm, oak, sycamore and mulberry, but methods for specific identification of a particular tree host species-limited strain or differentiation of tree-specific strains are lacking. It is also unknown whether a particular land...

  9. ISOLATION AND PHYLOGENETIC ANALYSIS OF XYLELLA FASTIDIOSA FROM ITS INVASIVE ALTERNATIVE HOST, PORCELAIN BERRY

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A strain of X. fastidiosa was isolated from an invasive alternative host species, porcelain berry. Its genetic relationship with strains isolated from a native alternative host, wild grape, and other economically important hosts including grape, peach, plum, oak, mulberry, maple and oleander was det...

  10. Microarray Analysis of Global Gene Expression of V. vinifera in Response to Xylella fastidiosa Infection

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Previously, we analyzed gene expression profiles of Pierce’s Disease (PD) resistant and susceptible genotypes of V. arizonica hybrids in response to infection by X. fastidiosa (Xf). Here we report the gene expression profile of the PD susceptible European grapevine (V. vinifera) in response to Xf...

  11. EFFECT OF THE GROWTH REGULATOR PACLOBUTRAZOL ON GROWTH OF THE BACTERIAL PATHOGEN XYLELLA FASTIDIOSA

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Xyella fastidiosa is a fastidious, xylem-limited, insect transmitted, bacterial plant pathogen that has a wide host range and causes bacterial leaf scorch (BLS) in shade trees. BLS is a chronic disorder characterized by late season leaf scorch and dieback and is common in urban and suburban areas o...

  12. Diffusible signal factor-repressed extracellular traits enable attachment of Xylella fastidiosa to insect vectors and transmission.

    PubMed

    Baccari, Clelia; Killiny, Nabil; Ionescu, Michael; Almeida, Rodrigo P P; Lindow, Steven E

    2014-01-01

    The hypothesis that a wild-type strain of Xylella fastidiosa would restore the ability of rpfF mutants blocked in diffusible signal factor production to be transmitted to new grape plants by the sharpshooter vector Graphocephala atropunctata was tested. While the rpfF mutant was very poorly transmitted by vectors irrespective of whether they had also fed on plants infected with the wild-type strain, wild-type strains were not efficiently transmitted if vectors had fed on plants infected with the rpfF mutant. About 100-fewer cells of a wild-type strain attached to wings of a vector when suspended in xylem sap from plants infected with an rpfF mutant than in sap from uninfected grapes. The frequency of transmission of cells suspended in sap from plants that were infected by the rpfF mutant was also reduced over threefold. Wild-type cells suspended in a culture supernatant of an rpfF mutant also exhibited 10-fold less adherence to wings than when suspended in uninoculated culture media. A factor released into the xylem by rpfF mutants, and to a lesser extent by the wild-type strain, thus inhibits their attachment to, and thus transmission by, sharpshooter vectors and may also enable them to move more readily through host plants. PMID:24571393

  13. Spatiotemporal colonization of Xylella fastidiosa in its vector supports the role of egestion in the inoculation mechanism of foregut-borne plant pathogens.

    PubMed

    Backus, Elaine A; Morgan, David J W

    2011-08-01

    The pathogen that causes Pierce's disease of grapevine, Xylella fastidiosa, is the only known bacterial, arthropod-transmitted plant pathogen that does not circulate in the vector's hemolymph. Instead, bacteria are foregut-borne, persistent in adult vectors but semipersistent in immatures (i.e., bacteria colonize cuticular surfaces of the anterior foregut, are retained for hours to days, but are lost during molting). Yet, exactly how a sharpshooter vector inoculates bacteria from foregut acquisition sites is unknown. The present study used confocal laser-scanning microscopy to identify locations in undissected, anterior foreguts of the glassy-winged sharpshooter colonized by green fluorescent protein-expressing X. fastidiosa. Spatial and temporal distributions of colonizing X. fastidiosa were examined daily over acquisition access periods of 1 to 6 days for both contaminated field-collected and clean laboratory-reared Homalodisca vitripennis. Results provide the first direct, empirical evidence that established populations of X. fastidiosa can disappear from vector foreguts over time. When combined with existing knowledge on behavior, physiology, and functional anatomy of sharpshooter feeding, present results support the idea that the disappearance is caused by outward fluid flow (egestion) not inward flow (ingestion) (i.e., swallowing). Thus, results support the hypothesis that egestion is a critical part of the X. fastidiosa inoculation mechanism. Furthermore, results suggest a cyclical, spatiotemporal pattern of microbial colonization, disappearance, and recolonization in the precibarium. Colonization patterns also support two types of egestion, termed rinsing and discharging egestion herein. Finally, comparison of acquisition results for field-collected versus laboratory-reared sharpshooters suggest that there may be competitive binding for optimum acquisition sites in the foregut. Therefore, successful inoculation of X. fastidiosa may depend, in large part, on

  14. Plant water stress effects on stylet probing behaviors of Homalodisca vitripennis (Hemiptera: Cicadellidae) associated with acquisition and inoculation of the bacterium Xylella fastidiosa.

    PubMed

    Krugner, Rodrigo; Backus, Elaine A

    2014-02-01

    ABSTRACT The glassy-winged sharpshooter, Homalodisca vitripennis (Germar) (Hemiptera: Cicadellidae), is a xylem fluid-ingesting leafhopper that transmits Xylella fastidiosa Wells et al., a plant-infecting bacterium that causes several plant diseases in the Americas. Although the role of plant water stress on the population density and dispersal ofH. vitripennis has been studied, nothing is known about the effects of plant water stress on the transmission of X. fastidiosa by H. vitripennis. A laboratory study was conducted to determine the influence of plant water stress on the sharpshooter stylet probing behaviors associated with the acquisition and inoculation of X. fastidiosa. Electrical penetration graph was used to monitor H. vitripennis feeding behaviors for 20-h periods on citrus [Citrus sinensis (L.) Osbeck] and almond [Prunus dulcis (Miller) D.A. Webb] plants subjected to levels of water stress. Adult H. vitripennis successfully located xylem vessels, then performed behaviors related to the evaluation of the xylem cell and fluid, and finally ingested xylem fluid from citrus and almond plants under the tested fluid tensions ranging from -5.5 to -33.0 bars and -6.0 to -24.5 bars, respectively. In general, long and frequent feeding events associated with the acquisition and inoculation of X. fastidiosa were observed only in fully irrigated plants (i.e., >-10 bars), which suggests that even low levels of plant water stress may reduce the spread of X. fastidiosa. Results provided insights to disease epidemiology and support the hypothesis that application of regulated deficit irrigation has the potential to reduce the incidence of diseases caused by X.fastidiosa by reducing the number of vectors and by decreasing pathogen transmission efficiency. PMID:24665686

  15. Cell Wall-Degrading Enzymes Enlarge the Pore Size of Intervessel Pit Membranes in Healthy and Xylella fastidiosa-Infected Grapevines1[C][W][OA

    PubMed Central

    Pérez-Donoso, Alonso G.; Sun, Qiang; Roper, M. Caroline; Greve, L. Carl; Kirkpatrick, Bruce; Labavitch, John M.

    2010-01-01

    The pit membrane (PM) is a primary cell wall barrier that separates adjacent xylem water conduits, limiting the spread of xylem-localized pathogens and air embolisms from one conduit to the next. This paper provides a characterization of the size of the pores in the PMs of grapevine (Vitis vinifera). The PM porosity (PMP) of stems infected with the bacterium Xylella fastidiosa was compared with the PMP of healthy stems. Stems were infused with pressurized water and flow rates were determined; gold particles of known size were introduced with the water to assist in determining the size of PM pores. The effect of introducing trans-1,2-diaminocyclohexane-N,N,N′,N′-tetraacetic acid (CDTA), oligogalacturonides, and polygalacturonic acid into stems on water flux via the xylem was also measured. The possibility that cell wall-degrading enzymes could alter the pore sizes, thus facilitating the ability of X. fastidiosa to cross the PMs, was tested. Two cell wall-degrading enzymes likely to be produced by X. fastidiosa (polygalactuoronase and endo-1,4- β -glucanase) were infused into stems, and particle passage tests were performed to check for changes in PMP. Scanning electron microscopy of control and enzyme-infused stem segments revealed that the combination of enzymes opened holes in PMs, probably explaining enzyme impacts on PMP and how a small X. fastidiosa population, introduced into grapevines by insect vectors, can multiply and spread throughout the vine and cause Pierce's disease. PMID:20107028

  16. Feeding Behaviors That Promote Inoculation Of Xylella fastidiosa Are Performed Less By Glassy-winged Sharpshooters on Resistant Vitis Candicans Than On Susceptible V. vinifera cv. ‘Chardonnay’

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Development of grape varieties resistant to Pierce’s Disease, caused by the lethal bacterium Xylella fastidiosa (Xf), is considered the most sustainable, long-term solution to the disease. Grape breeders are working to develop varieties resistant to multiplication and spread of the bacterium, by in...

  17. Directed evolution induces tributyrin hydrolysis in a virulence factor of Xylella fastidiosa using a duplicated gene as a template.

    PubMed

    Gouran, Hossein; Chakraborty, Sandeep; Rao, Basuthkar J; Asgeirsson, Bjarni; Dandekar, Abhaya

    2014-01-01

    Duplication of genes is one of the preferred ways for natural selection to add advantageous functionality to the genome without having to reinvent the wheel with respect to catalytic efficiency and protein stability. The duplicated secretory virulence factors of Xylella fastidiosa (LesA, LesB and LesC), implicated in Pierce's disease of grape and citrus variegated chlorosis of citrus species, epitomizes the positive selection pressures exerted on advantageous genes in such pathogens. A deeper insight into the evolution of these lipases/esterases is essential to develop resistance mechanisms in transgenic plants. Directed evolution, an attempt to accelerate the evolutionary steps in the laboratory, is inherently simple when targeted for loss of function. A bigger challenge is to specify mutations that endow a new function, such as a lost functionality in a duplicated gene. Previously, we have proposed a method for enumerating candidates for mutations intended to transfer the functionality of one protein into another related protein based on the spatial and electrostatic properties of the active site residues (DECAAF). In the current work, we present in vivo validation of DECAAF by inducing tributyrin hydrolysis in LesB based on the active site similarity to LesA. The structures of these proteins have been modeled using RaptorX based on the closely related LipA protein from Xanthomonas oryzae. These mutations replicate the spatial and electrostatic conformation of LesA in the modeled structure of the mutant LesB as well, providing in silico validation before proceeding to the laborious in vivo work. Such focused mutations allows one to dissect the relevance of the duplicated genes in finer detail as compared to gene knockouts, since they do not interfere with other moonlighting functions, protein expression levels or protein-protein interaction. PMID:25717364

  18. Grapevine rootstock effects on scion sap phenolic levels, resistance to Xylella fastidiosa infection, and progression of Pierce's disease

    PubMed Central

    Wallis, Christopher M.; Wallingford, Anna K.; Chen, Jianchi

    2013-01-01

    The xylem-limited bacterium Xylella fastidiosa (Xf) causes Pierce's disease (PD), an important disease of grapevine, Vitis vinifera L. Grapevine rootstocks were developed to provide increased resistance to root disease, but rootstock effects on cane and vine diseases remain unclear. Grapevines that consisted of Cabernet Sauvignon or Chardonnay grafted to 13 different rootstocks were inoculated with Xf and evaluated for PD severity and Xf titer after 6 months. A subset of six rootstock/scion combinations had xylem sap phenolic levels assessed in non-infected and Xf-infected grapevines. Vigor also was analyzed by measuring root lengths and masses. Cabernet Sauvignon grafted to 101-14MG, 1103P, 420A, or Schwarzmann had reduced PD severity compared to Cabernet Sauvignon grafted to 110R, 5BB, or SO4. Chardonnay grafted to Salt Creek or Freedom had reduced PD severity compared to Chardonnay grafted to RS3 or Schwarzmann. Chardonnay grafted to RS3 had greater Xf titer than Chardonnay grafted to 101-14MG, Freedom, or Salt Creek. No other differences in Xf titer among rootstocks were observed. Of the six scion/rootstock combinations which had xylem sap phenolics analyzed, Chardonnay/RS3 had the highest levels of most phenolics whereas Cabernet Sauvignon/101-14MG had the lowest phenolic levels. However, Chardonnay/101-14MG, which had mild PD symptoms, had greater sap levels of caftaric acid than other scion/rootstock combinations. Sap levels of caftaric acid, methyl salicylate, a procyanidin trimer, and quinic acid were greater in Xf-infected vs. non-infected grapevines. Chardonnay on 101-14MG or Salt Creek had greater root mass than Chardonnay on RS3. Cabernet Sauvignon on 101-14MG had greater root mass than Cabernet Sauvignon on 110R. These results identified rootstocks with the capacity for reducing PD symptom progression. Rootstocks also were shown to affect Xf titer, xylem sap phenolic levels, and plant vigor. PMID:24376452

  19. The MqsRA Toxin-Antitoxin System from Xylella fastidiosa Plays a Key Role in Bacterial Fitness, Pathogenicity, and Persister Cell Formation

    PubMed Central

    Merfa, Marcus V.; Niza, Bárbara; Takita, Marco A.; De Souza, Alessandra A.

    2016-01-01

    Through the formation of persister cells, bacteria exhibit tolerance to multidrug and other environmental stresses without undergoing genetic changes. The toxin-antitoxin (TA) systems are involved in the formation of persister cells because they are able to induce cell dormancy. Among the TA systems, the MqsRA system has been observed to be highly induced in persister cells of Xylella fastidiosa (causal agent of citrus variegated chlorosis—CVC) activated by copper stress, and has been described in Escherichia coli as related to the formation of persister cells and biofilms. Thus, we evaluated the role of this TA system in X. fastidiosa by overexpressing the MqsR toxin, and verified that the toxin positively regulated biofilm formation and negatively cell movement, resulting in reduced pathogenicity in citrus plants. The overexpression of MqsR also increased the formation of persister cells under copper stress. Analysis of the gene and protein expression showed that this system likely has an autoregulation mechanism to express the toxin and antitoxin in the most beneficial ratio for the cell to oppose stress. Our results suggest that this TA system plays a key role in the adaptation and survival of X. fastidiosa and reveal new insights into the physiology of phytopathogen-host interactions. PMID:27375608

  20. Characterization of the TolB-Pal trans-envelope complex from Xylella fastidiosa reveals a dynamic and coordinated protein expression profile during the biofilm development process.

    PubMed

    Santos, Clelton A; Janissen, Richard; Toledo, Marcelo A S; Beloti, Lilian L; Azzoni, Adriano R; Cotta, Monica A; Souza, Anete P

    2015-10-01

    The intriguing roles of the bacterial Tol-Pal trans-envelope protein complex range from maintenance of cell envelope integrity to potential participation in the process of cell division. In this study, we report the characterization of the XfTolB and XfPal proteins of the Tol-Pal complex of Xylella fastidiosa. X. fastidiosa is a major plant pathogen that forms biofilms inside xylem vessels, triggering the development of diseases in important cultivable plants around the word. Based on functional complementation experiments in Escherichia coli tolB and pal mutant strains, we confirmed the role of xftolB and xfpal in outer membrane integrity. In addition, we observed a dynamic and coordinated protein expression profile during the X. fastidiosa biofilm development process. Using small-angle X-ray scattering (SAXS), the low-resolution structure of the isolated XfTolB-XfPal complex in solution was solved for the first time. Finally, the localization of the XfTolB and XfPal polar ends was visualized via immunofluorescence labeling in vivo during bacterial cell growth. Our results highlight the major role of the components of the cell envelope, particularly the TolB-Pal complex, during the different phases of bacterial biofilm development. PMID:26049080

  1. Identification of a response regulator involved in surface attachment, cell-cell aggregation, exopolysaccharide production and virulence in the plant pathogen Xylella fastidiosa.

    PubMed

    Voegel, Tanja M; Doddapaneni, Harshavardhan; Cheng, Davis W; Lin, Hong; Stenger, Drake C; Kirkpatrick, Bruce C; Roper, M Caroline

    2013-04-01

    Xylella fastidiosa, the causal agent of Pierce's disease of grapevine, possesses several two-component signal transduction systems that allow the bacterium to sense and respond to changes in its environment. Signals are perceived by sensor kinases that autophosphorylate and transfer the phosphate to response regulators (RRs), which direct an output response, usually by acting as transcriptional regulators. In the X. fastidiosa genome, 19 RRs were found. A site-directed knockout mutant in one unusual RR, designated XhpT, composed of a receiver domain and a histidine phosphotransferase output domain, was constructed. The resulting mutant strain was analysed for changes in phenotypic traits related to biofilm formation and gene expression using microarray analysis. We found that the xhpT mutant was altered in surface attachment, cell-cell aggregation, exopolysaccharide (EPS) production and virulence in grapevine. In addition, this mutant had an altered transcriptional profile when compared with wild-type X. fastidiosa in genes for several biofilm-related traits, such as EPS production and haemagglutinin adhesins. PMID:23186359

  2. The MqsRA Toxin-Antitoxin System from Xylella fastidiosa Plays a Key Role in Bacterial Fitness, Pathogenicity, and Persister Cell Formation.

    PubMed

    Merfa, Marcus V; Niza, Bárbara; Takita, Marco A; De Souza, Alessandra A

    2016-01-01

    Through the formation of persister cells, bacteria exhibit tolerance to multidrug and other environmental stresses without undergoing genetic changes. The toxin-antitoxin (TA) systems are involved in the formation of persister cells because they are able to induce cell dormancy. Among the TA systems, the MqsRA system has been observed to be highly induced in persister cells of Xylella fastidiosa (causal agent of citrus variegated chlorosis-CVC) activated by copper stress, and has been described in Escherichia coli as related to the formation of persister cells and biofilms. Thus, we evaluated the role of this TA system in X. fastidiosa by overexpressing the MqsR toxin, and verified that the toxin positively regulated biofilm formation and negatively cell movement, resulting in reduced pathogenicity in citrus plants. The overexpression of MqsR also increased the formation of persister cells under copper stress. Analysis of the gene and protein expression showed that this system likely has an autoregulation mechanism to express the toxin and antitoxin in the most beneficial ratio for the cell to oppose stress. Our results suggest that this TA system plays a key role in the adaptation and survival of X. fastidiosa and reveal new insights into the physiology of phytopathogen-host interactions. PMID:27375608

  3. A SYBR green-based real-time polymerase chain reaction protocol and novel DNA extraction technique to detect Xylella fastidiosa in Homalodisca coagulata.

    PubMed

    Bextine, Blake; Blua, Matthew; Harshman, Dave; Miller, Thomas A

    2005-06-01

    Homalodisca coagulata Say (Hemiptera: Cicadellidae) is a major agronomic pest because it transmits Xylella fastidiosa (Wells), the bacterium that causes Pierce's disease of grapevine. The ability to easily detect X. fastidiosa in populations of H. coagulata facilitates epidemiological studies and development of a monitoring program supporting disease management. Such a program depends on a detection protocol that is rapid, reproducible, and amenable to large sample sizes, while remaining sensitive enough to detect low amounts of pathogen DNA. In this study, we developed an improved method to speed DNA extraction by implementing a simple vacuum step that replaces labor- and time-intensive maceration of tissue samples and that is compatible with manufactured DNA extraction kits. Additionally, we have developed a SYBR Green-based real-time (RT)-polymerase chain reaction (PCR) system, which uses traditional PCR primers that are relatively inexpensive and effective. Using this extraction/RT-PCR system, we found no statistically significant differences in the detection of X. fastidiosa among samples that were either immediately extracted or stored dry or in mineral oil for 10 d at -4 degrees C. In further testing, we found no significant reduction in detection capabilities for X. fastidiosa-fed H. coagulata left in the sun on yellow sticky cards for up to 6 d. Therefore, we recommend a field-based detection system that includes recovery of H. coagulata from sticky traps for up to 6 d after trapping, subsequent freezing of samples for as long as 10 d before vacuum extraction is performed, and detection of the bacterium by SYBR Green-based RT-PCR. PMID:16022291

  4. Assessing Adhesion Forces of Type I and Type IV Pili of Xylella fastidiosa Bacteria by Use of a Microfluidic Flow Chamber▿ †

    PubMed Central

    De La Fuente, Leonardo; Montanes, Emilie; Meng, Yizhi; Li, Yaxin; Burr, Thomas J.; Hoch, H. C.; Wu, Mingming

    2007-01-01

    Xylella fastidiosa, a bacterium responsible for Pierce's disease in grapevines, possesses both type I and type IV pili at the same cell pole. Type IV pili facilitate twitching motility, and type I pili are involved in biofilm development. The adhesiveness of the bacteria and the roles of the two pili types in attachment to a glass substratum were evaluated using a microfluidic flow chamber in conjunction with pilus-defective mutants. The average adhesion force necessary to detach wild-type X. fastidiosa cells was 147 ± 11 pN. Mutant cells possessing only type I pili required a force of 204 ± 22 pN for removal, whereas cells possessing only type IV pili required 119 ± 8 pN to dislodge these cells. The experimental results demonstrate that microfluidic flow chambers are useful and convenient tools for assessing the drag forces necessary for detaching bacterial cells and that with specific pilus mutants, the role of the pilus type can be further assessed. PMID:17293518

  5. A rhamnose-rich O-antigen mediates adhesion, virulence, and host colonization for the xylem-limited phytopathogen Xylella fastidiosa.

    PubMed

    Clifford, Jennifer C; Rapicavoli, Jeannette N; Roper, M Caroline

    2013-06-01

    Xylella fastidiosa is a gram-negative, xylem-limited bacterium that causes a lethal disease of grapevine called Pierce's disease. Lipopolysaccharide (LPS) composes approximately 75% of the outer membrane of gram-negative bacteria and, because it is largely displayed on the cell surface, it mediates interactions between the bacterial cell and its surrounding environment. LPS is composed of a conserved lipid A-core oligosaccharide component and a variable O-antigen portion. By targeting a key O-antigen biosynthetic gene, we demonstrate the contribution of the rhamnose-rich O-antigen to surface attachment, cell-cell aggregation, and biofilm maturation: critical steps for successful infection of the host xylem tissue. Moreover, we have demonstrated that a fully formed O-antigen moiety is an important virulence factor for Pierce's disease development in grape and that depletion of the O-antigen compromises its ability to colonize the host. It has long been speculated that cell-surface polysaccharides play a role in X. fastidiosa virulence and this study confirms that LPS is a major virulence factor for this important agricultural pathogen. PMID:23441576

  6. N-acetylcysteine in agriculture, a novel use for an old molecule: focus on controlling the plant-pathogen Xylella fastidiosa.

    PubMed

    Muranaka, Lígia S; Giorgiano, Thais E; Takita, Marco A; Forim, Moacir R; Silva, Luis F C; Coletta-Filho, Helvécio D; Machado, Marcos A; de Souza, Alessandra A

    2013-01-01

    Xylella fastidiosa is a plant pathogen bacterium that causes diseases in many different crops. In citrus, it causes Citrus Variegated Chlorosis (CVC). The mechanism of pathogenicity of this bacterium is associated with its capacity to colonize and form a biofilm in the xylem vessels of host plants, and there is not yet any method to directly reduce populations of this pathogen in the field. In this study, we investigated the inhibitory effect of N-Acetylcysteine (NAC), a cysteine analogue used mainly to treat human diseases, on X. fastidiosa in different experimental conditions. Concentrations of NAC over 1 mg/mL reduced bacterial adhesion to glass surfaces, biofilm formation and the amount of exopolysaccharides (EPS). The minimal inhibitory concentration of NAC was 6 mg/mL. NAC was supplied to X. fastidiosa-infected plants in hydroponics, fertigation, and adsorbed to organic fertilizer (NAC-Fertilizer). HPLC analysis indicated that plants absorbed NAC at concentrations of 0.48 and 2.4 mg/mL but not at 6 mg/mL. Sweet orange plants with CVC symptoms treated with NAC (0.48 and 2.4 mg/mL) in hydroponics showed clear symptom remission and reduction in bacterial population, as analyzed by quantitative PCR and bacterial isolation. Experiments using fertigation and NAC-Fertilizer were done to simulate a condition closer to that normally is used in the field. For both, significant symptom remission and a reduced bacterial growth rate were observed. Using NAC-Fertilizer the lag for resurgence of symptoms on leaves after interruption of the treatment increased to around eight months. This is the first report of the anti-bacterial effect of NAC against a phytopathogenic bacterium. The results obtained in this work together with the characteristics of this molecule indicate that the use of NAC in agriculture might be a new and sustainable strategy for controlling plant pathogenic bacteria. PMID:24009716

  7. Crystallization and preliminary X-ray analysis of BigR, a transcription repressor from Xylella fastidiosa involved in biofilm formation

    SciTech Connect

    Barbosa, Rosicler Lázaro; Rinaldi, Fábio Cupri; Guimarães, Beatriz Gomes Benedetti, Celso Eduardo

    2007-07-01

    In order to gain new insights into the protein structure and its possible interaction with a metal ion or effector ligand, BigR from X. fastidiosa was crystallized in native and selenomethionine (SeMet) labelled forms using the hanging-drop vapour-diffusion method. BigR (biofilm growth-associated repressor) is a novel repressor protein that regulates the transcription of an operon implicated in biofilm growth in both Xylella fastidiosa and Agrobacterium tumefaciens. This protein binds to a palindromic TA-rich element located in the promoter of the BigR operon and strongly represses transcription of the operon. BigR contains a helix–turn–helix (HTH) domain that is found in some members of the ArsR/SmtB family of metal sensors, which control metal resistance in bacteria. Although functional studies have suggested that BigR does not act as a metal sensor, the presence of two cysteines and a methionine in its primary structure raised the possibility of BigR being a metal-ligand protein. In order to gain new insights into the protein structure and its possible interaction with a metal ion or effector ligand, BigR from X. fastidiosa was crystallized in native and selenomethionine (SeMet) labelled forms using the hanging-drop vapour-diffusion method. X-ray diffraction data were collected from native and SeMet crystals to resolutions of 1.95 and 2.2 Å, respectively. Both crystals belong to space group P321 and contain one molecule per asymmetric unit.

  8. Sequence/structural analysis of xylem proteome emphasizes pathogenesis-related proteins, chitinases and β-1, 3-glucanases as key players in grapevine defense against Xylella fastidiosa

    PubMed Central

    Chakraborty, Sandeep; Nascimento, Rafael; Zaini, Paulo A.; Gouran, Hossein; Rao, Basuthkar J.; Goulart, Luiz R.

    2016-01-01

    Background. Xylella fastidiosa, the causative agent of various plant diseases including Pierce’s disease in the US, and Citrus Variegated Chlorosis in Brazil, remains a continual source of concern and economic losses, especially since almost all commercial varieties are sensitive to this Gammaproteobacteria. Differential expression of proteins in infected tissue is an established methodology to identify key elements involved in plant defense pathways. Methods. In the current work, we developed a methodology named CHURNER that emphasizes relevant protein functions from proteomic data, based on identification of proteins with similar structures that do not necessarily have sequence homology. Such clustering emphasizes protein functions which have multiple copies that are up/down-regulated, and highlights similar proteins which are differentially regulated. As a working example we present proteomic data enumerating differentially expressed proteins in xylem sap from grapevines that were infected with X. fastidiosa. Results. Analysis of this data by CHURNER highlighted pathogenesis related PR-1 proteins, reinforcing this as the foremost protein function in xylem sap involved in the grapevine defense response to X. fastidiosa. β-1, 3-glucanase, which has both anti-microbial and anti-fungal activities, is also up-regulated. Simultaneously, chitinases are found to be both up and down-regulated by CHURNER, and thus the net gain of this protein function loses its significance in the defense response. Discussion. We demonstrate how structural data can be incorporated in the pipeline of proteomic data analysis prior to making inferences on the importance of individual proteins to plant defense mechanisms. We expect CHURNER to be applicable to any proteomic data set. PMID:27257535

  9. Infectivity and transmission of Xylella fastidiosa Salento strain by Philaenus spumarius L. (Hemiptera: Aphrophoridae) in Apulia, Italy

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Discovery of X. fastidiosa from olive trees with “Olive quick decline syndrome" (OQDS) in October 2013 on the western coast of the Salento Peninsula prompted an immediate search for insect vectors of the bacterium. The dominant xylem-fluid feeding hemipteran collected in olive orchards was the meado...

  10. Combined use of a new SNP-based assay and multilocus SSR markers to assess genetic diversity of Xylella fastidiosa subsp. pauca infecting citrus and coffee plants.

    PubMed

    Montes-Borrego, Miguel; Lopes, Joao R S; Jiménez-Díaz, Rafael M; Landa, Blanca B

    2015-03-01

    Two haplotypes of Xylella fastidiosa subsp. pauca (Xfp) that correlated with their host of origin were identified in a collection of 90 isolates infecting citrus and coffee plants in Brazil, based on a single-nucleotide polymorphism in the gyrB sequence. A new single-nucleotide primer extension (SNuPE) protocol was designed for rapid identification of Xfp according to the host source. The protocol proved to be robust for the prediction of the Xfp host source in blind tests using DNA from cultures of the bacterium, infected plants, and insect vectors allowed to feed on Xfp-infected citrus plants. AMOVA and STRUCTURE analyses of microsatellite data separated most Xfp populations on the basis of their host source, indicating that they were genetically distinct. The combined use of the SNaPshot protocol and three previously developed multilocus SSR markers showed that two haplotypes and distinct isolates of Xfp infect citrus and coffee in Brazil and that multiple, genetically different isolates can be present in a single orchard or infect a single tree. This combined approach will be very useful in studies of the epidemiology of Xfp-induced diseases, host specificity of bacterial genotypes, the occurrence of Xfp host jumping, vector feeding habits, etc., in economically important cultivated plants or weed host reservoirs of Xfp in Brazil and elsewhere. PMID:26415663

  11. The Type II Secreted Lipase/Esterase LesA is a Key Virulence Factor Required for Xylella fastidiosa Pathogenesis in Grapevines

    PubMed Central

    Nascimento, Rafael; Gouran, Hossein; Chakraborty, Sandeep; Gillespie, Hyrum W.; Almeida-Souza, Hebréia O.; Tu, Aye; Rao, Basuthkar J.; Feldstein, Paul A.; Bruening, George; Goulart, Luiz R.; Dandekar, Abhaya M.

    2016-01-01

    Pierce’s disease (PD) of grapevines is caused by Xylella fastidiosa (Xf), a xylem-limited gamma-proteobacterium that is responsible for several economically important crop diseases. The occlusion of xylem elements and interference with water transport by Xf and its associated biofilm have been posited as the main cause of PD symptom development; however, Xf virulence mechanisms have not been described. Analysis of the Xf secretome revealed a putative lipase/esterase (LesA) that was abundantly secreted in bacterial culture supernatant and was characterized as a protein ortholog of the cell wall-degrading enzyme LipA of Xanthomonas strains. LesA was secreted by Xf and associated with a biofilm filamentous network. Additional proteomic analysis revealed its abundant presence in outer membrane vesicles (OMVs). Accumulation of LesA in leaf regions associated positively with PD symptoms and inversely with bacterial titer. The lipase/esterase also elicited a hypersensitive response in grapevine. Xf lesA mutants were significantly deficient for virulence when mechanically inoculated into grapevines. We propose that Xf pathogenesis is caused by LesA secretion mediated by OMV cargos and that its release and accumulation in leaf margins leads to early stages of observed PD symptoms. PMID:26753904

  12. The Type II Secreted Lipase/Esterase LesA is a Key Virulence Factor Required for Xylella fastidiosa Pathogenesis in Grapevines.

    PubMed

    Nascimento, Rafael; Gouran, Hossein; Chakraborty, Sandeep; Gillespie, Hyrum W; Almeida-Souza, Hebréia O; Tu, Aye; Rao, Basuthkar J; Feldstein, Paul A; Bruening, George; Goulart, Luiz R; Dandekar, Abhaya M

    2016-01-01

    Pierce's disease (PD) of grapevines is caused by Xylella fastidiosa (Xf), a xylem-limited gamma-proteobacterium that is responsible for several economically important crop diseases. The occlusion of xylem elements and interference with water transport by Xf and its associated biofilm have been posited as the main cause of PD symptom development; however, Xf virulence mechanisms have not been described. Analysis of the Xf secretome revealed a putative lipase/esterase (LesA) that was abundantly secreted in bacterial culture supernatant and was characterized as a protein ortholog of the cell wall-degrading enzyme LipA of Xanthomonas strains. LesA was secreted by Xf and associated with a biofilm filamentous network. Additional proteomic analysis revealed its abundant presence in outer membrane vesicles (OMVs). Accumulation of LesA in leaf regions associated positively with PD symptoms and inversely with bacterial titer. The lipase/esterase also elicited a hypersensitive response in grapevine. Xf lesA mutants were significantly deficient for virulence when mechanically inoculated into grapevines. We propose that Xf pathogenesis is caused by LesA secretion mediated by OMV cargos and that its release and accumulation in leaf margins leads to early stages of observed PD symptoms. PMID:26753904

  13. Quantification and localization of hesperidin and rutin in Citrus sinensis grafted on C. limonia after Xylella fastidiosa infection by HPLC-UV and MALDI imaging mass spectrometry.

    PubMed

    Soares, Márcio Santos; da Silva, Danielle Fernandes; Forim, Moacir Rossi; da Silva, Maria Fátima das Graças Fernandes; Fernandes, João Batista; Vieira, Paulo Cezar; Silva, Denise Brentan; Lopes, Norberto Peporine; de Carvalho, Sérgio Alves; de Souza, Alessandra Alves; Machado, Marcos Antônio

    2015-07-01

    A high performance liquid chromatography-ultraviolet (HPLC-UV) method was developed for quantifying hesperidin and rutin levels in leaves and stems of Citrus limonia, with a good linearity over a range of 1.0-80.0 and 1.0-50.0 μg mL(-1) respectively, with r(2)>0.999 for all curves. The limits of detection (LOD) for both flavonoids were 0.6 and 0.5 μg mL(-1), respectively, with quantification (LOQ) being 2.0 and 1.0 μg mL(-1), respectively. The quantification method was applied to Citrus sinensis grafted onto C. limonia with and without CVC (citrus variegated chlorosis) symptoms after Xylella fastidiosa infection. The total content of rutin was low and practically constant in all analyses in comparison with hesperidin, which showed a significant increase in its amount in symptomatic leaves. Scanning electron microscopy studies on leaves with CVC symptoms showed vessel occlusion by biofilm, and a crystallized material was noted. Considering the difficulty in isolating these crystals for analysis, tissue sections were analyzed by matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI MSI) to confirm the presence of hesperidin at the site of infection. The images constructed from MS/MS data with a specific diagnostic fragment ion (m/z 483) also showed higher ion intensities for it in infected plants than in healthy ones, mainly in the vessel regions. These data suggest that hesperidin plays a role in the plant-pathogen interaction, probably as a phytoanticipin. This method was also applied to C. sinensis and C. limonia seedlings, and comparison with the graft results showed that the rootstock had an increased hesperidin content ∼3.6 fold greater in the graft stem than in the stem of C. sinensis seedlings. Increase in hesperidin content by rootstock can be related to induced internal defense mechanisms. PMID:25749617

  14. Phenotype overlap in Xylella fastidiosa is controlled by the cyclic di-GMP phosphodiesterase Eal in response to antibiotic exposure and diffusible signal factor-mediated cell-cell signaling.

    PubMed

    de Souza, Alessandra A; Ionescu, Michael; Baccari, Clelia; da Silva, Aline M; Lindow, Steven E

    2013-06-01

    Eal is an EAL domain protein in Xylella fastidiosa homologous to one involved in resistance to tobramycin in Pseudomonas aeruginosa. EAL and HD-GYP domain proteins are implicated in the hydrolysis of the secondary messenger bis-(3'-5')-cyclic dimeric GMP (cyclic di-GMP). Cell density-dependent communication mediated by a Diffusible Signal Factor (DSF) also modulates cyclic di-GMP levels in X. fastidiosa, thereby controlling the expression of virulence genes and genes involved in insect transmission. The possible linkage of Eal to both extrinsic factors such as antibiotics and intrinsic factors such as quorum sensing, and whether both affect virulence, was thus addressed. Expression of eal was induced by subinhibitory concentrations of tobramycin, and an eal deletion mutant was more susceptible to this antibiotic than the wild-type strain and exhibited phenotypes similar to those of an rpfF deletion mutant blocked in DSF production, such as hypermotility, reduced biofilm formation, and hypervirulence to grape. Consistent with that, the rpfF mutant was more susceptible than the wild-type strain to tobramycin. Therefore, we propose that cell-cell communication and antibiotic stress can apparently lead to similar modulations of cyclic di-GMP in X. fastidiosa, resulting in similar phenotypes. However, the effect of cell density is dominant compared to that of antibiotic stress, since eal is suppressed by RpfF, which may prevent inappropriate behavioral changes in response to antibiotic stress when DSF accumulates. PMID:23542613

  15. Enhanced Reliability and Accuracy for Field Deployable Bioforensic Detection and Discrimination of Xylella fastidiosa subsp. pauca, Causal Agent of Citrus Variegated Chlorosis Using Razor Ex Technology and TaqMan Quantitative PCR

    PubMed Central

    Fletcher, Jacqueline; Melcher, Ulrich; Ochoa Corona, Francisco Manuel

    2013-01-01

    A reliable, accurate and rapid multigene-based assay combining real time quantitative PCR (qPCR) and a Razor Ex BioDetection System (Razor Ex) was validated for detection of Xylella fastidiosa subsp. pauca (Xfp, a xylem-limited bacterium that causes citrus variegated chlorosis [CVC]). CVC, which is exotic to the United States, has spread through South and Central America and could significantly impact U.S. citrus if it arrives. A method for early, accurate and sensitive detection of Xfp in plant tissues is needed by plant health officials for inspection of products from quarantined locations, and by extension specialists for detection, identification and management of disease outbreaks and reservoir hosts. Two sets of specific PCR primers and probes, targeting Xfp genes for fimbrillin and the periplasmic iron-binding protein were designed. A third pair of primers targeting the conserved cobalamin synthesis protein gene was designed to detect all possible X. fastidiosa (Xf) strains. All three primer sets detected as little as 1 fg of plasmid DNA carrying X. fastidiosa target sequences and genomic DNA of Xfp at as little as 1 - 10 fg. The use of Razor Ex facilitates a rapid (about 30 min) in-field assay capability for detection of all Xf strains, and for specific detection of Xfp. Combined use of three primer sets targeting different genes increased the assay accuracy and broadened the range of detection. To our knowledge, this is the first report of a field-deployable rapid and reliable bioforensic detection and discrimination method for a bacterial phytopathogen based on multigene targets. PMID:24312333

  16. Structures of the N-acetyltransferase domain of Xylella fastidiosa N-acetyl-l-glutamate synthase/kinase with and without a His tag bound to N-acetyl-l-glutamate

    PubMed Central

    Zhao, Gengxiang; Jin, Zhongmin; Allewell, Norma M.; Tuchman, Mendel; Shi, Dashuang

    2015-01-01

    Structures of the catalytic N-acetyltransferase (NAT) domain of the bifunctional N-acetyl-l-glutamate synthase/kinase (NAGS/K) from Xylella fastidiosa bound to N-acetyl-l-glutamate (NAG) with and without an N-terminal His tag have been solved and refined at 1.7 and 1.4 Å resolution, respectively. The NAT domain with an N-terminal His tag crystallized in space group P41212, with unit-cell parameters a = b = 51.72, c = 242.31 Å. Two subunits form a molecular dimer in the asymmetric unit, which contains ∼41% solvent. The NAT domain without an N-terminal His tag crystallized in space group P21, with unit-cell parameters a = 63.48, b = 122.34, c = 75.88 Å, β = 107.6°. Eight subunits, which form four molecular dimers, were identified in the asymmetric unit, which contains ∼38% solvent. The structures with and without the N-terminal His tag provide an opportunity to evaluate how the His tag affects structure and function. Furthermore, multiple subunits in different packing environments allow an assessment of the plasticity of the NAG binding site, which might be relevant to substrate binding and product release. The dimeric structure of the X. fastidiosa N-acetytransferase (xfNAT) domain is very similar to that of human N-acetyltransferase (hNAT), reinforcing the notion that mammalian NAGS is evolutionally derived from bifunctional bacterial NAGS/K. PMID:25615976

  17. Structures of the N-acetyltransferase domain of Xylella fastidiosa N-acetyl-L-glutamate synthase/kinase with and without a His tag bound to N-acetyl-L-glutamate.

    PubMed

    Zhao, Gengxiang; Jin, Zhongmin; Allewell, Norma M; Tuchman, Mendel; Shi, Dashuang

    2015-01-01

    Structures of the catalytic N-acetyltransferase (NAT) domain of the bifunctional N-acetyl-L-glutamate synthase/kinase (NAGS/K) from Xylella fastidiosa bound to N-acetyl-L-glutamate (NAG) with and without an N-terminal His tag have been solved and refined at 1.7 and 1.4 Å resolution, respectively. The NAT domain with an N-terminal His tag crystallized in space group P4(1)2(1)2, with unit-cell parameters a=b=51.72, c=242.31 Å. Two subunits form a molecular dimer in the asymmetric unit, which contains ∼41% solvent. The NAT domain without an N-terminal His tag crystallized in space group P21, with unit-cell parameters a=63.48, b=122.34, c=75.88 Å, β=107.6°. Eight subunits, which form four molecular dimers, were identified in the asymmetric unit, which contains ∼38% solvent. The structures with and without the N-terminal His tag provide an opportunity to evaluate how the His tag affects structure and function. Furthermore, multiple subunits in different packing environments allow an assessment of the plasticity of the NAG binding site, which might be relevant to substrate binding and product release. The dimeric structure of the X. fastidiosa N-acetytransferase (xfNAT) domain is very similar to that of human N-acetyltransferase (hNAT), reinforcing the notion that mammalian NAGS is evolutionally derived from bifunctional bacterial NAGS/K. PMID:25615976

  18. A TaqMan-based real time PCR assay for specific detection and quantification of Xylella fastidiosa strains causing bacterial leaf scorch in oleander

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A TaqMan-based real-time PCR assay is developed for strains of X. fastidiosa causing oleander leaf scorch. The assay uses primers WG-OLS-F1 and WG-OLS-R1 and the fluorescent probe WG-OLS-P1, designed based on unique sequences present only in genomic sequence of oleander strain Ann1. The assay is spe...

  19. Xylella taiwanensis sp. nov. cause of pear leaf scorch disease in Taiwan

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Xylella fastidiosa is a group of xylem-limited and nutritionally fastidious plant pathogenic bacteria. While mostly found in the Americas, new X. fastidiosa strains have been reported from other continents such as Asia, including a pear leaf scorch (PLS) strain from Taiwan. Current taxonomy of X. fa...

  20. Assessment of the importance of alfalfa to the epidemiology of xylellae diseases in the San Joaquin Valley of California

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The role of alfalfa in the epidemiology of xylellae diseases in the San Joaquin Valley of California was assessed. Alfalfa was investigated as it is a known host of Xylella fastidiosa and often harbors large populations of a native vector, Draeculacephala minerva. Laboratory inoculation of fourtee...

  1. First report of Xylella fastidiosa in Avocado

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Since the late 1990s, chlorotic mottling, marginal scorch, deformation of leaves, defoliation, shortening of internodes, and branch dieback have been observed in avocado trees (Persea americana Mill.) in Costa Rica. The symptoms are not uniformly distributed in the tree, so some branches of a tree ...

  2. MOTILITY TRAILS OF XYLELLA FASTDIOSA ON CULTURE MEDIA SURFACES VISUALIZED BY REFLECTION CONTRAST MICROSCOPY.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Xylella fastidiosa is an important plant pathogen causing grape Pierce’s disease (PD) and almond leaf scorch disease (ALSD) in California. Because of its nutritional fastidiousness, few biological traits of this bacterium have been reported. In this study, two ALSD strains, M12 and M23, representing...

  3. Epidemiology of xylellae diseases in the San Joaquin Valley of California: the role of alfalfa

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Pierce’s disease of grape and almond leaf scorch disease are both caused by various stains of the bacterial pathogen Xylella fastidiosa. The pathogen is vectored by xylem feeding insects. Within the San Joaquin Valley of California, the green sharpshooter (Draeculacephala Minerva) is one of the mos...

  4. THE ROLE OF GLASSY-WINGED SHARPSHOOTER SALIVARY ENZYMES IN INFECTION AND MOVEMENT OF X. FASTIDIOSA

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The purpose of this project is to determine whether a vector’s enzymatic saliva aids the establishment of the few ‘pioneer’ Xylella fastidiosa (Xf) cells that are inoculated into a plant; thus the bacteria would co-localize with the saliva. Previous work showed that watery saliva of glassy-winged ...

  5. Use of GIS maps in assessing the role of alfalfa in the epidemiology of xylellae diseases in California

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Grape and almond are planted throughout much of California. Both crops are susceptible to diseases caused by Xylella fastidiosa. Within California’s Central Valley, the green sharpshooter (Draeculacephala minerva) is a key vector. This insect is often found in large numbers in cultivated alfalfa ...

  6. TOWARDS THE DEVELOPMENT OF A SEQUENCE-BASED PCR SYSTEM FOR DETECTION AND GENOMIC STUDY OF XYLELLA FASTIDOSA STRAINS IMPORTANT TO CALIFORNIA

    Technology Transfer Automated Retrieval System (TEKTRAN)

    We report two of our recent efforts for improvement of PCR detection and genomic analysis of Xylella fastidiosa. We evaluated the use of PCR primers from a hyper-variable locus to monitor diversity within strains of the same pathotype. The pspB (PD1208) locus, encoding a serine protease, was select...

  7. Differences in Stylet Penetration Behaviors of Glassy-winged Sharpshooters on Xylella-Resistant Vitis candicans vs. Susceptible Vitis vinifera cv. ‘Chardonnay’

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Electrical penetration graph (EPG) monitoring was used to compare stylet penetration behaviors of glassy-winged sharpshooter (GWSS), a vector of Xylella fastidiosa (Xf), on Xf-resistant Vitis candicans grape vs. susceptible V. vinifera cv. ‘Chardonnay.’ Frequency of occurrence of X waves (represent...

  8. Spatiotemporal colonization of Xyllela fastidiosa in its vector supports two types of egestion in the inoculation mechanism of foregut-borne plant pathogens

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The bacterial agent that causes Pierce’s disease of grapevine, Xylella fastidiosa, is the only known arthropod-transmitted prokaryotic plant pathogen that does not circulate in the vector’s hemolymph. Instead, bacteria are foregut-borne and semi-persistent, i.e. bacteria colonize cuticular surface...

  9. Beta 1, 4-glucanase in Glassy-Winged Sharpshooter Saliva, and its Possible Role in Infection and Movement of X. fastidiosa

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The purpose of this project is to determine whether 1, 4-glucanase (EGase), the major enzymatic protein in watery saliva of glassy-winged sharpshooter (GWSS) co-localizes via immunocytochemistry with the few ‘pioneer’ Xylella fastidiosa (Xf) cells that are inoculated into a plant by this vector. If...

  10. Infectivity and transmission of Xylellua fastidiosa by Philaenus spumarius (Hemiptera: Aphrophoridae) in Apulia, Italy.

    PubMed

    Saponari, Maria; Loconsole, Giuliana; Cornara, Daniele; Yokomi, Raymond K; De Stradis, Angelo; Boscia, Donato; Bosco, Domenico; Martelli, Giovanni P; Krugner, Rodrigo; Porcelli, Francesco

    2014-08-01

    Discovery of Xylella fastidiosa from olive trees with "Olive quick decline syndrome" in October 2013 on the west coast of the Salento Peninsula prompted an immediate search for insect vectors of the bacterium. The dominant xylem-fluid feeding hemipteran collected in olive orchards during a 3-mo survey was the meadow spittlebug, Philaenus spumarius (L.) (Hemiptera: Aphrophoridae). Adult P. spumarius, collected in November 2013 from ground vegetation in X. fastidiosa-infected olive orchards, were 67% (40 out of 60) positive for X. fastidiosa by polymerase chain reaction (PCR) assays. Euscelis lineolatus Brullé were also collected but tested negative for the pathogen. Transmission tests with P. spumarius collected from the Salento area were, therefore, conducted. After a 96-h inoculation access period with 8 to 10 insects per plant and a 30-d incubation period, PCR results showed P. spumarius transmitted X. fastidiosa to two of five periwinkle plants but not to the seven olive plants. Sequences of PCR products from infected periwinkle were identical with those from X. fastidiosa-infected field trees. These data showed P. spumarius as a vector of X. fastidiosa strain infecting olives trees in the Salento Peninsula, Italy. PMID:25195417

  11. THE CONTRIBUTION OF THE PECTIN-DEGRADING ENZYME POLYGALACTURONASE (PG) IN TRANSMISSION OF X. FASTIDIOSA TO GRAPE AND THE USE OF PG-INHIBITING PROTEINS FOR TRANSGENIC RESISTANCE TO PIERCE'S DISEASE

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Work this year has shown that polygalacturonase (PG) of Xylella fastidiosa (Xf) is an endo-acting enzyme. This provides a reference that can now be used to biochemically characterize the interaction between the pathogen’s PG and PG inhibitor proteins (PGIPs) that is implied by the fact that transg...

  12. Small RNAs in Xylella fastidiosa and their epidemiological applications

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Bacterial non-coding small RNAs (sRNAs) have attracted considerable attention due to their roles in regulating numerous cellular processes including survival, adaptation and pathogenesis. Sequence variation in sRNA genes reflect a previously unrecognized source of genomic diversity in bacteria. Xy...

  13. NEW STANDARD PROTOCOL FOR DIAGNOSIS OF XYLELLA FASTIDIOSA

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The Interim Commission on Phytosanitary Measures of the International Plant Protection Convention (IPPC) adopted recommendations on the publication of International Standards for Phytosanitary Measures (ISPM). This guideline produces standardized documents describing procedures and methods for the d...

  14. Mutations in Type I and Type IV Pilus Biosynthetic Genes Affect Twitching Motility Rates in Xylella fastidiosa▿ †

    PubMed Central

    De La Fuente, Leonardo; Burr, Thomas J.; Hoch, Harvey C.

    2007-01-01

    Xylella fastidiosa possesses both type I and type IV pili at the same cell pole. By use of a microfluidic device, the speed of twitching movement by wild-type cells on a glass surface against the flow direction of media was measured as 0.86 (standard error [SE], 0.04) μm min−1. A type I pilus mutant (fimA) moved six times faster (4.85 [SE, 0.27] μm min−1) and a pilY1 mutant moved three times slower (0.28 [SE, 0.03] μm min−1) than wild-type cells. Type I pili slow the rate of movement, while the putative type IV pilus protein PilY1 is likely important for attachment to surfaces. PMID:17693510

  15. PCR screening for Xylella fastidiosa in grape genebank accessions collected in the Southeast US.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The National Clonal Germplasm Repository (NCGR) in Davis, CA maintains the US collection of warm climate grapes in a field genebank in Winters, CA. This collection includes numerous accessions which are highly susceptible to Pierce’s Disease (PD) as well as many PD-tolerant accessions which are nat...

  16. Transcriptional Regulation of Grape Cytochrome P450 Gene Expression in Response to Xylella fastidiosa

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Plant cytochrome P450 monooxygenases are versatile redox proteins that mediate biosynthesis of lignins, terpenes, alkaloids, and a variety of other secondary compounds as plant defense agents against a range of pathogens and insects. To determine if cytochrome P450 monooxygenases are involved in the...

  17. A toxin-antitoxin system encoded by the Xylella fastidiosa chromosome regulates growth

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Bacteria encode toxin-antitoxin (TA) systems consisting of a stable toxin and a cognate labile antitoxin. When encoded by a plasmid, TA systems confer stable plasmid inheritance. When encoded by the chromosome, TA systems may confer advantageous responses to environmental stress. The chromosome of...

  18. Two whole genome sequences of Xylella fastidiosa almond leaf scorch strains

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Almond leaf scorch disease (ALSD) is currently re-emerging in the Central Valley of California, threatening the production of this high value crop. Our knowledge about biology and pathogenicity of ALSD is currently limited, hindering the development of effective disease control strategies. ALSD is c...

  19. Feeding Behaviors of the Glassy-Winged Sharpshooter that Control Inoculation of Xylella fastidiosa

    Technology Transfer Automated Retrieval System (TEKTRAN)

    In the final eight months of this grant, we completed remaining studies and emphasized writing results for publication. Overall, all objectives of this grant were successfully completed. The work identified the electrical penetration graph (EPG) waveforms responsible for both ingestion and egestio...

  20. Transcriptional Regulation of the Grape Cytochrome P450 Monooxygenase Gene CYP736B Expression in Response to Xylella fastidiosa Infection

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Plant cytochrome P450 monooxygenases are a group of versatile redox proteins that mediate the biosynthesis of lignins, terpenes, alkaloids, and a variety of other secondary compounds which act as plant defense agents. To determine if cytochrome P450 monooxygenases are involved in defense response to...

  1. Susceptibility to Xylella fastidiosa in a first generation hybrid from a non-traditional peach-almond cross

    Technology Transfer Automated Retrieval System (TEKTRAN)

    To facilitate development of Prunus rootstocks with desirable agronomic traits, domesticated peach (Prunus persica) and almond (P. dulcis) were crossed with wild almond relatives. This work reports that a hybrid from a P. webbii x P. persica cv Harrow Blood cross is susceptible to almond leaf scorch...

  2. Salivary enzymes are injected into xylem by the glassy-winged sharpshooter, a vector of Xylella fastidiosa

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Certain hemipteran insects such as the glassy-winged sharpshooter, Homalodisca vitripennis, subsist entirely on xylem fluid, notwithstanding the poor nutrition of such food. Among many adaptations enabling xylem-feeding are aspects of the insect’s salivation that may also allow these insects to tra...

  3. Filamentation and spatiotemporal distribution of extracellular polymeric substances: role on X.fastidiosa single cell adhesion and biofilm formation (Conference Presentation)

    NASA Astrophysics Data System (ADS)

    Janissen, Richard; Murillo, Duber M.; Niza, Barbara; Sahoo, Prasana K.; Monteiro, Moniellen P.; César, Carlos L.; Carvalho, Hernandes F.; de Souza, Alessandra A.; Cotta, Monica A.

    2016-04-01

    Biofilms can be defined as a community of microorganisms attached to a surface, living embedded in a self- produced matrix of hydrated extracellular polymeric substances (EPS) which comprises most of the biofilm mass. We have recently used an extensive pool of microscopy techniques (confocal fluorescence, electron and scanning probe microscopies) at the micro and nanoscales in order to create a detailed temporal observation of Xylella fastidiosa biofilm formation, using both wild type strain and Green Fluorescent Protein (GFP)-modified cells of this citrus phytopathogen. We have identified three different EPS compositions, as well as their spatial and temporal distribution from single cell to mature biofilm formation stages. In the initial adhesion stage, soluble-EPS (S-EPS) accumulates at cell polar regions and forms a surface layer which facilitates irreversible cell attachment and cell cluster formation. These small clusters are subsequently connected by filamentous cells; further S-EPS surface coverage facilitates cell attachment and form filaments, leading to a floating framework of mature biofilms. The important role of EPS in X.fastidiosa biology was further investigated by imunolabelling experiments to detect the distribution of XadA1 adhesin, which is expressed in early stages of biofilm formation and released in outer membrane vesicles. This protein is located mainly in S-EPS covered areas, as well as on the filaments, indicating a molecular pathway to the enhanced cell attachment previously observed. These results suggest that S-EPS may thus represent an important target for disease control, slow plant colonization by the bacteria, keeping the plant more productive in the field.

  4. Emendation of the Genus Actinokineospora Hasegawa 1988 and Transfer of Amycolatopsis fastidiosa Henssen et al. 1987 as Actinokineospora fastidiosa comb. nov

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The species Amycolatopsis fastidiosa was proposed by Henssen et al. (1987), based on morphological and chemotaxonomic observations, for a strain originally described as 'Pseudonocardia fastidiosa' by Celmer et al. in a US Patent issued in 1977. In the course of a phylogenetic study of the valid tax...

  5. Xylem structure of four grape varieties and 12 alternative hosts to the xylem-limited bacterium Xylella fastidious

    PubMed Central

    Chatelet, David S.; Wistrom, Christina M.; Purcell, Alexander H.; Rost, Thomas L.; Matthews, Mark A.

    2011-01-01

    Background and Aims The bacterium Xylella fastidiosa (Xf), responsible for Pierce's disease (PD) of grapevine, colonizes the xylem conduits of vines, ultimately killing the plant. However, Vitis vinifera grapevine varieties differ in their susceptibility to Xf and numerous other plant species tolerate Xf populations without showing symptoms. The aim of this study was to examine the xylem structure of grapevines with different susceptibilities to Xf infection, as well as the xylem structure of non-grape plant species that support or limit movement of Xf to determine if anatomical differences might explain some of the differences in susceptibility to Xf. Methods Air and paint were introduced into leaves and stems to examine the connectivity between stem and leaves and the length distribution of their vessels. Leaf petiole and stem anatomies were studied to determine the basis for the free or restricted movement of Xf into the plant. Key Results There were no obvious differences in stem or petiole vascular anatomy among the grape varieties examined, nor among the other plant species that would explain differences in resistance to Xf. Among grape varieties, the more tolerant ‘Sylvaner’ had smaller stem vessel diameters and 20 % more parenchyma rays than the other three varieties. Alternative hosts supporting Xf movement had slightly longer open xylem conduits within leaves, and more connection between stem and leaves, when compared with alternative hosts that limit Xf movement. Conclusions Stem–leaf connectivity via open xylem conduits and vessel length is not responsible for differences in PD tolerance among grape varieties, or for limiting bacterial movement in the tolerant plant species. However, it was found that tolerant host plants had narrower vessels and more parenchyma rays, possibly restricting bacterial movement at the level of the vessels. The implications of xylem structure and connectivity for the means and regulation of bacterial movement are

  6. WHERE, WHEN AND HOW DO INGESTION AND OTHER FEEDING BEHAVIORS OF THE GLASSY-WINGED SHARPSHOOTER ALLOW INOCULATION OF XYLELLA FASTIDIOSA

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This year, we completed the laboratory portion of work that identified an aspect of electrical penetration graph (EPG) ingestion waveform (C) of glassy-winged sharpshooter (GWSS) that definitively represents xylem ingestion. We also demonstrated that extravasation is correlated with the B1 waveform...

  7. Sharpshooter X-wave: Correlation of an Electrical Penetration Graph (EPG) Waveform With Xylem Penetration Supports a Hypothesized Mechanism For Xylella Fastidiosa Inoculation

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Electrical Penetration Graph (EPG) monitoring is the most rigorous means of observation and quantification of feeding by piercing-sucking arthropods. Previous EPG studies with aphids and leafhoppers have demonstrated that the X waveform identifies when the insect is ingesting from its preferred pla...

  8. Effects of kaolin particle film and imidacloprid on glassy-winged sharpshooter (Homalodisca vitripennis) (Hemiptera: Cicadellidae)populations and the prevention of spread of Xylella fastidiosa in grape

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The glassy-winged sharpshooter (GWSS), Homalodisca coagulata (Say), was introduced into California and soon became a major pest of important agronomic, horticultural, landscape, ornamental crops and native trees in California. This pest feeds readily on grape and, in doing so, transmits X. fastidio...

  9. Investigation on solubilization protocols in the refolding of the thioredoxin TsnC from Xylella fastidiosa by high hydrostatic pressure approach.

    PubMed

    Lemke, Laura Simoni; Chura-Chambi, Rosa Maria; Rodrigues, Daniella; Cussiol, Jose Renato Rosa; Malavasi, Natalia Vallejo; Alegria, Thiago Geronimo Pires; Netto, Luis Eduardo Soares; Morganti, Ligia

    2015-02-01

    The lack of efficient refolding methodologies must be overcome to take full advantage of the fact that bacteria express high levels of aggregated recombinant proteins. High hydrostatic pressure (HHP) impairs intermolecular hydrophobic and electrostatic interactions, dissociating aggregates, which makes HHP a useful tool to solubilize proteins for subsequent refolding. A process of refolding was set up by using as a model TsnC, a thioredoxin that catalyzes the disulfide reduction to a dithiol, a useful indication of biological activity. The inclusion bodies (IB) were dissociated at 2.4 kbar. The effect of incubation of IB suspensions at 1-800 bar, the guanidine hydrochloride concentration, the oxidized/reduced glutathione (GSH/GSSG) ratios, and the additives in the refolding buffer were analyzed. To assess the yields of fully biologically active protein obtained for each tested condition, it was crucial to analyze both the TsnC solubilization yield and its enzymatic activity. Application of 2.4 kbar to the IB suspension in the presence of 9 mM GSH, 1mM GSSG, 0.75 M guanidine hydrochloride, and 0.5M arginine with subsequent incubation at 1 bar furnished high refolding yield (81%). The experience gained in this study shall help to establish efficient HHP-based protein refolding processes for other proteins. PMID:25448595

  10. AN EVOLUTIONARY PERSPECTIVE OF XYELLAE DISEASES IN GRAPEVIN, CITRUS AND MULBERRY

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Xylella fastidiosa causes diseases on many economically important plants. An understanding of how xylellae diseases originated and evolved is interesting as well as important for disease prevention and management. We evaluated the phylogenetic relationships among strains from citrus, grapevine, and ...

  11. Interactions of citrus variegated chlorosis (CVC) with endophytic bacteria

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Citrus variegated chlorosis (CVC), is a disease of sweet orange [Citrus sinensis (L.)], is caused by Xylella fastidiosa subsp. pauca, a phytopathogenic bacterium that has been shown to infect all sweet orange cultivars. Xylella fastidiosa is a fastidious Gram negative, xylem-limited bacterium which ...

  12. Non contiguous-finished genome sequence and description of Dielma fastidiosa gen. nov., sp. nov., a new member of the Family Erysipelotrichaceae

    PubMed Central

    Ramasamy, Dhamodharan; Lagier, Jean-Christophe; Nguyen, Thi Tien; Raoult, Didier; Fournier, Pierre-Edouard

    2013-01-01

    Dielma fastidiosa strain JC13T gen. nov., sp. nov. is the type strain of D. fastidiosa gen. nov., sp. nov., the type species of a new genus within the family Erysipelotrichaceae. This strain, whose draft genome is described here, was isolated from the fecal flora of a healthy 16-year-old male Senegalese volunteer. D. fastidiosa is a Gram-negative anaerobic rod. Here we describe the features of this organism, together with the complete genome sequence and annotation. The 3,574,031 bp long genome comprises a 3,556,241-bp chromosome and a 17,790-bp plasmid. The chromosome contains 3,441 protein-coding and 50 RNA genes, including 3 rRNA genes, whereas the plasmid contains 17 protein-coding genes. PMID:23991263

  13. Effects of citrus and avocado irrigation and nitrogen-form soil amendment on host selection by adult Homalodisca vitripennis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Host plant water status is thought to influence dispersal and movement of Homalodisca vitripennis Germar, especially where plants are grown under high evaporative demand. The leafhopper transmits the bacterial pathogen, Xylella fastidiosa Wells et. al, to economically important susceptible plants. ...

  14. GENE EXPRESSION IN TWO LEAFHOPPER VECTORS OF PIERCES DISEASE OF GRAPES, GLASSY-WINGED SHARPSHOOTER AND BLUE-GREEN SHARPSHOOTER (HEMIPTERA: CICADELLIDAE)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Production of a unigene set identified the putative functions of over 350 transcripts from two leafhopper species (Hemiptera: Cicadellidae) which vector the plant infecting bacterium, Xylella fastidiosa. Genetic similarities between the glassy-winged sharpshooter, Homalodisca coagulata, and the blue...

  15. Plant water stress effects on the net dispersal rate of the insect vector Homalodisca vitripennis (Germar) (Hemiptera: Cicadellidae) and movement of its egg parasitoid, Gonatocerus ashmeadi Girault (Hymenoptera: Mymaridae)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Homalodisca vitripennis, one of the main vectors of Xylella fastidiosa, is associated with citrus plantings in California, USA. Infested citrus orchards act as a source of vectors to adjacent vineyards where X. fastidiosa causes Pierce’s disease (PD). An analysis of the pattern and rate of movement ...

  16. Rootstock effects on almond leaf scorch disease incidence and severity

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A five-year field study was conducted to evaluate effects of duration and exclusion of Xylella fastidiosa infections on young almond tree performance and their links to tree vigor. ‘Nemaguard’, ‘Okinawa’, ‘Nonpareil’, and Y119 were used as rootstocks for almond scion ‘Sonora’. Among X.fastidiosa-inf...

  17. Salivary Secretions of the Glassy-winged Sharpshooter, Homalodisca vitripennis: Novel Method of Collection and Protein Profile

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The glassy-winged sharpshooter, Homalodisca vitripennis Takiya (Hemiptera: Cicadellidae) is a xylophagous leafhopper that efficiently transmits Xylella fastidiosa, causative agent of a variety of scorch diseases such as Pierce’s Disease of grapes. The transmission mechanism of X. fastidiosa by the ...

  18. Exploring glassy-winged sharpshooter microbiota using deep 16S rRNA sequencing from individual insects

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The glassy-winged sharpshooter (GWSS) is an invasive insect species that transmits Xylella fastidiosa, the bacterium causing Pierce’s disease of grapevine and other leaf scorch diseases. X. fastidiosa has been shown to colonize the anterior foregut (cibarium and precibarium) of sharpshooters, where ...

  19. Glassy-winged sharpshooter Microbiota explored using deep 16S rRNA sequencing from individual insects

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The glassy-winged sharpshooter (GWSS) is an invasive insect species that transmits Xylella fastidiosa, the bacterium causing Pierce’s disease of grapevine and other leaf scorch diseases. X. fastidiosa has been shown to colonize the anterior foregut (cibarium and precibarium) of sharpshooters, where ...

  20. Competitive binding influences Xf vector load: Confocal and SEM images of GFP-expressing Xf in GWSS foreguts

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Like all hemipteran vectors of plant pathogens, glassy-winged sharpshooters, Homalodisca vitripennis acquire their load of Xylella fastidiosa (Xf ) via ingestion of fluids containing bacteria. X. fastidiosa then colonizes the anterior regions of the vector’s foregut, the cibarium (or sucking pump) ...

  1. Anterior foregut microbiota of the glassy-winged sharpshooter explored using deep 16S rRNA gene sequencing from individual insects

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The glassy-winged sharpshooter (GWSS) is an invasive insect species that transmits Xylella fastidiosa, the bacterium causing Pierce’s disease of grapevine and other leaf scorch diseases. X. fastidiosa has been shown to colonize the anterior foregut (cibarium and precibarium) of sharpshooters, where ...

  2. Population genomic analysis of a bacterial plant pathogen: Novel insight into the origin of Pierce's disease of grapevine in the U.S.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Pierce’s disease of grape (PD) has long posed a serious threat to the wine industry in the United States. It is caused by infection of xylem vessels by a subspecific form of Xylella fastidiosa (subsp. fastidiosa) defined by a genetically distinct clade at least 15,000 years old (Schuenzel et al. 200...

  3. Glassy-winged sharpshooter oviposition effects on foliar grapevine and red-tipped photinia terpenoid levels

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The glassy-winged sharpshooter (GWSS), Homalodisca vitripennis (Germar) (Hemiptera: Cicadellidae), is an important vector of Xylella fastidiosa, the bacterium that causes Pierce's disease of grapevine and is a threat to grape production throughout the United States. Female GWSS deposit egg masses be...

  4. Glassy-winged sharpshooter oviposition effects on photinia volatile chemistry with implications on egg parasitoid effectiveness

    Technology Transfer Automated Retrieval System (TEKTRAN)

    An effective way to limit incidence of Pierce’s disease of grapevine is to reduce populations of glassy-winged sharpshooter (GWSS), Homalodisca vitripennis (Germar) (Hemiptera: Cicadellidae), which transmit the causal bacterium, Xylella fastidiosa. One strategy is to utilize egg parasitoids such as ...

  5. Metabolic Profiling of Xylem Sap from Pierce’s Disease Resistant and Susceptible Grapevines

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Pierce’s Disease (PD) of grapevines is caused by a gram-negative, xylem-limited bacterium Xylella fastidiosa (Xf). All Vitis vinifera-based cultivars are highly susceptible to Xf infection. However, some grape species from the southern United States such as V. arizonica, V. Shuttleworthii, and Musca...

  6. Transmission of bacterial plant pathogens by Hemipteran vectors: The intersection of genomics and classical vector biology

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Insects that are classified in the Order Hemiptera have become one of the most important plant-associated insect taxa, mainly because of their ability to transmit plant pathogens. The glassy-winged sharpshooter is the primary vector of the plant-infecting bacterium Xylella fastidiosa, the causal age...

  7. Using Marker-Assisted Selection to Breed Pierce’s Disease-Resistant Grapes

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Pierce’s disease (PD), caused by the bacterium Xylella fastidiosa, is an important disease of grapevines in California, across the southern United States, and into South America. In regions where disease pressure is high, cultivation of Vitis vinifera cultivars is difficult or impossible. This study...

  8. Molecular evidence for three novel viruses in Glassy-winged Sharpshooters, 'Homalidisca vitripennis' (Hemiptera: Cicadellidae)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A metagenomic approach was used to identify viral pathogens in the glassy-winged sharpshooter, GWSS, Homalodisca vitripennis (=H. coagulata), is renowned as the principal vector of the plant pathogenic bacterium Xylella fastidiosa which causes a number of economically important diseases including Pi...

  9. Evaluation and Molecular Characterization of Beauveria bassiana for the Control of the Glassy-winged Sharpshooter (Homoptera: Cicadellidae) in California.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The glassy-winged sharpshooter, Homalodisca coagulata (Say), is an important pest on grapes, citrus, almonds and other commercial crops in California as it is a vector of Xylella fastidiosa Wells, a bacterium that causes Pierce’s disease in grapes, citrus variegated chlorosis, almond leaf scorch and...

  10. EXPERIMENTAL ANALYSIS OF BIOLOGICAL PARAMETERS AND VECTOR ABILITY OF GLASSY-WINGED SHARPSHOOTERS FROM ALLOPATRIC POPULATIONS IN CALIFORNIA

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The glassy-winged sharpshooter (GWSS), Homalodisca vitripennis (Germar), is native to the southeastern United States and northeastern Mexico. It was detected in southern California in the late 1980s and in the San Joaquin Valley in 1999, where it transmits the bacterium Xylella fastidiosa to grapev...

  11. Electrical Penetration Graph (EPG) monitoring of feeding by glassy-winged sharpshooter and potential application of EPG to other pathogen vectors

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Recent research on feeding of the glassy-winged sharpshooter, Homalodisca vitripennis (Germar) and the exact mechanism of its transmission (acquisition and inoculation) of Xylella fastidiosa (Xf) to grape will be used as an example of the utility of a new, universal, AC-DC Electrical Penetration Gra...

  12. CORRELATIONS OF CIBARIAL AND PRECIBARIAL MUSCLE ACTIVITIES WITH EPG WAVEFORMS AND EXCRETION IN THE SMOKE-TREE SHARPSHOOTER, HOMALODISCA LITURATA (HEMIPTERA: CICADELLIDAE)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The smoke-tree sharpshooter, Homalodisca liturata, like its close relative, the glassy-winged sharpshooter, H. coagulata, is a generalist xylophagous leafhopper known to be a vector of Xylella fastidiosa. The stylet penetration behavior of sharpshooters in plants is most rigorously studied via elect...

  13. Propagation of Homalodisca Coagulata Virus-01 via Homalodisca Vitripennis cell culture

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The glassy-winged sharpshooter (Homalodisca vitripennis) is a highly vagile and polyphagous insect found throughout the southwestern United States. These insects are the predominant vectors of Xylella fastidiosa, a xylem-limited bacterium that is the causal agent of Pierce's disease (PD) of grapevin...

  14. Effects of Almond Leaf Scorch Disease on Almond Yield and Implications for Management

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Almond leaf scorch (ALS) disease has been present in California’s almond growing regions for over 60 years. This disease is caused by the bacterium Xylella fastidiosa and the pathogen is vectored by xylem feeding sharpshooters and spittlebugs. Currently there are no effective management techniques...

  15. Abundance and consumption rate of glassy-winged sharpshooter (Hemiptera: Cicadellidae) on peaches and plums

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Homalodisca vitripennis, also known as the glassy-winged sharpshooter, is a primary vector of phony peach and plum leaf scald diseases caused by Xylella fastidiosa Wells et al. Two of the following scions, (Prunus persica L. Batch cvs. Flordaking and June Gold and Prunus salicina L. cvs. Methley an...

  16. Citrus Viruses in Guatemala: Application of Laboratory-Based Assays

    Technology Transfer Automated Retrieval System (TEKTRAN)

    In preparation for a citrus certification in Guatemala, there was an urgent need to determine which graft transmissible citrus pathogens were present. Because of the lack of biological indicator plants, Citrus tristeza virus (CTV) and Xylella fastidiosa, causal agent for citrus variegated chlorosis...

  17. Seasonal abundance of sharpshooters, leafhoppers, and psyllids associated with potatoes affected by zebra chip disorder

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A survey of psyllids, leafhoppers, and sharpshooters in commercial potatoes near McAllen, TX in the Lower Rio Grande Valley was conducted using yellow sticky traps. Species from these groups that were collected in the survey were evaluated for the presence of Xylella fastidiosa, the suspected causa...

  18. Phylogenetic analysis of heat shock proteins in Glassy-winged sharpshooter

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Four heat shock protein transcripts were produced from the glassy-winged sharpshooter Homalodisca vitripennis (Germar) (Hemiptera: Cicadellidae) which is the major vector of Xylella fastidiosa, the causal agent of Pierce’s disease of grapes. As genomic information has continued to be produced resea...

  19. Glassy-winged sharpshooters (GWSS) are not GWSS: Differential Reproductive Maturity between Allopatric Californian Populations

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Homalodisca vitripennis (Germar) is native to southeastern U.S. and northeastern Mexico. It was detected in southern California in the late 1980s and in the San Joaquin Valley in 1999, where it transmits the bacterium Xylella fastidiosa Wells et al. to grapevines and other crops. The reproductive ...

  20. Sequential sampling plans for estimating censity of glassy-winged sharpshooter, homalodisca vitripennis (Hemiptera: Cicadellidae) on citrus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The glassy-winged sharpshooter (GWSS), Homalodisca vitripennis, is a serious pest of grapes and other crop and ornamental plants mainly through its role as a vector of the bacterium Xylella fastidiosa. Citrus harbors large populations of GWSS throughout much of the year in areas where the pest is pr...

  1. Attraction of the egg parasitoid, Gonatocerus ashmeadi Girault (Hymenoptera: Mymaridae) to synthetic formulation of a (E)-ß-ocimene and (E,E)-a-farnesene mixture

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Glassy-winged sharpshooter (GWSS), Homalodisca vitripennis (Germar), is a vector of the xylem-inhabitant bacterium Xylella fastidiosa Wells et al., which causes Pierce’s disease of grapevines. Current GWSS control strategies are limited to area-wide insecticide applications and mass release of egg ...

  2. Effect of host plant fertilization on the developmental biology and feeding preference of the glassy-winged sharpshooter.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The main objective of this research was to evaluate the effect of host plant fertilization on the survival, immature development, adult fecundity, and the feeding of the glassy-winged sharpshooter (GWSS), a primary vector of Xylella fastidiosa (Xf). The development biology of GWSS was studied on co...

  3. Quantitative comparison of stylet penetration behaviors of glassy-winged sharpshooter, Homalodisca vitripennis, on four crop plants important in New Zealand and the USA

    Technology Transfer Automated Retrieval System (TEKTRAN)

    New Zealand is threatened by invasion of the glassy-winged sharpshooter (GWSS), Homalodisca vitripennis, an important vector of Xylella fastidiosa, a gram-negative bacterium that causes Pierce’s Disease in grape and scorch diseases in many other horticultural crops. Therefore, an understanding of t...

  4. Epidemiology of almond leaf scorch disease in the San Joaquin Valley of California

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Almond leaf scorch (ALS) disease has been present in California for more than 60 years. This disease is caused by the bacterium Xylella fastidiosa, which causes several other important plant diseases, including Pierce’s disease of grapes. The epidemiology of ALS in the San Joaquin Valley of Califo...

  5. Gene expression in adult blue-green sharpshooters, Graphocephala atropunctata (Signoret) (Hemiptera: Cicadellidae)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    We provide valuable, new information on the genetic basis for the biology of the adult blue-green sharpshooters, BGSS, Graphocephala atropunctata, which transmit the plant pathogenic bacterium, Xylella fastidiosa, the cause of Pierce’s disease of grapes, and other ‘Scorch-like’ diseases of tree crop...

  6. Sequence polymorphism of a glassy-winged sharpshooter phytoreovirus reveals a bottleneck in the Californian population

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The glassy-winged sharpshooter (GWSS; Homalodisca vitripennis Germar) is an invasive insect introduced to California circa 1989. Native to the southeastern U.S. and northeastern Mexico, GWSS is of economic concern as a vector of the Pierce’s disease bacterium Xylella fastidiosa. Recently, a novel ...

  7. Suitability of fertilized and non-fertilized eggs of Homalodisca vitripennis for the egg parasitoid Gonatocerus morrili

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The glassy-winged sharpshooter (GWSS), Homalodisca vitripennis, is a key vector of Xylella fastidiosa, the bacterium that causes Pierce’s disease of grapevines and other diseases of high-valued crops in the Americas. GWSS invaded California in the late 1980s and continues to threaten the $4 billion ...

  8. Sequential Sampling Plans for Estimating Density of Glassy-winged Sharpshooter, Homalodisca vitripennis (Hemiptera: Cicadellidae) on Citrus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The glassy-winged sharpshooter (GWSS), Homalodisca vitripennis, is a serious pest of grapes and other crop and ornamental plants mainly through its role as a vector of the bacterium Xylella fastidiosa. Citrus harbors large populations of GWSS throughout much of the year in areas where the pest is pr...

  9. Glassy-winged sharpshooter infestations induce grape hosts to produce volatile compounds which may attract egg parasitoids

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Natural predators or parasitoids may be attracted to prey by specific volatiles emitted from prey-infested host plants. Glassy-winged sharpshooter (GWSS), Homalodisca vitripennis (Germar), is a pest of particular concern as a vector of the Pierce’s disease pathogen, Xylella fastidiosa. GWSS egg para...

  10. Immunological detection of glassy-winged sharpshooter saliva in grapevine

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Glassy-winged sharpshooter (GWSS), Homalodisca vitripennis, is a major vector for transmission of Xylella fastidiosa (Xf), the causative agent of Pierce’s Disease in grapevine. During the feeding process of stylet penetration and xylem fluid ingestion, GWSS inject saliva into the plant. Inoculation...

  11. COMPARATIVE PROTEOMIC ANALYSIS OF STEM TISSUE AND XYLEM SAP FROM PIERCE'S DISEASE RESISTANT AND SUSCEPTIBLE GRAPEVINES

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Analyses of host plant resistance / susceptibility mechanisms to Xylella fastidiosa (Xf) infection are critical for understanding host-pathogen interactions. Proteomic analyses of stem tissue and xylem sap samples were initiated to complement genomic approaches employed in elucidating Pierce’s Dise...

  12. Management of almond leaf scorch disease: long term data on yield, tree vitality, and disease progress

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Almond leaf scorch (ALS) disease has been a chronic problem for California almond growers. This disease is caused by the bacterial pathogen Xylella fastidiosa and is transmitted by xylem-feeding insects. Previous research suggested that retaining, rather than roguing, ALS-affected trees may be more ...

  13. Long term evaluation of the effects of almond leaf scorch disease on orchard productivity

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Almond leaf scorch (ALS) disease has been present in California for more than 60 years. This disease is caused by the bacterium Xylella fastidiosa, which also causes several other important plant diseases, most notably Pierce’s disease of grapes. With the exception of planting resistant cultivars...

  14. Egg load dynamics of Homalodisca vitripennis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Homalodisca vitripennis, the glassy-winged sharpshooter, poses a serious threat to grape production due to its ability to vector Xylella fastidiosa, the causal agent of Pierce’s disease. The glassy-winged sharpshooter is native to the Southeastern United States and over the last twenty years has ex...

  15. Estimation of feeding threshold for Homalodisca vitripennis (Hemiptera: Cicadellidae) and its application to prediction of overwintering mortality

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The glassy-winged sharpshooter, Homalodisca vitripennis (Germar), is an invasive species that vectors the bacterium Xylella fastidiosa that induces Pierce’s Disease of grape. This study determined the effect of temperature on the feeding activity of H. vitripennis adults measured as the amount of e...

  16. Diverse niche opportunities for glassy-winged sharpshooter, Homalodisca vitripennis (Germar) (Hemiptera: Cicadellidae), in California and the role of research for crop protection

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Glassy-winged sharpshooter (GWSS) is a xylem feeder that vectors Xylella fastidiosa, a bacterium that causes Pierce’s Disease (PD) in grapevines and other diseases in a number of economically important plant species. GWSS originally from southeastern North America invaded California sometime in the ...

  17. Identification and whole extraction of Homalodisca coagulata Virus 01 (HoCv-01)from Texas Glassy-winged sharpshooter populations

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Viral infection makes insects more sensitive to insecticide treatment, resulting in lower LD50 rates needed to achieve significant control. The glassy-winged sharpshooter, GWSS, Homalodisca vitripennis (Hemiptera: Cicadellidae), is an invasive pest and important vector of Xylella fastidiosa, a xylem...

  18. Development and use of recombinant Homalodisca coagulata Virus-1 for controlling Homalodisca vitripennis, the glassy-winged sharpshooter

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The glassy-winged sharpshooter (GWSS; Homalodisca vitripennis) is an efficient vector of Xylella fastidiosa, causal agent of Pierce’s disease of grapevines. The goal of this research is to generate recombinant insect viruses to deliver toxic peptides and/or induce systemic RNA interference (RNAi) an...

  19. Grape hosts infested with glassy-winged sharpshooters produce volatile compounds which may attract egg parasitoids

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Glassy-winged sharpshooter (GWSS), Homalodisca vitripennis (Germar), is an important vector of Xylella fastidiosa Wells, which causes Pierce’s disease in grapes. Current management strategies in GWSS infested areas include mass release of the egg parasitoid Gonatocerus ashmeadi Girault and related s...

  20. VECTOR ABILITY AND REPRODUCTIVE CHARACTERISTICS OF HOMALODISCA VITRIPENNIS (GERMAR) (HEMIPTERA: CICADELLIDAE) FROM GEOGRAPHICALLY SEPARATED POPULATIONS IN CALIFORNIA

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The glassy-winged sharpshooter (GWSS), Homalodisca vitripennis (Germar), is native to the southeastern United States and northeastern Mexico. It was detected in southern California in the late 1980s and in the San Joaquin Valley in 1999, where it transmits the bacterium Xylella fastidiosa to grapev...

  1. Immunohistochemistry of ß 1,4-glucanase, the major enzymatic component of glassy-winged sharpshooter saliva, in probed grape petioles

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The purpose of this project is to determine whether ß 1, 4-glucanase (EGase), the major enzyme in the saliva of glassy-winged sharpshooter (GWSS), co-localizes with the few ‘pioneer’ Xylella fastidiosa (Xf) cells that are inoculated into a plant by this vector. If it does, then this suggests that ve...

  2. Expressed sequence tags from the black-winged sharpshooter: Application to biology and vector control

    Technology Transfer Automated Retrieval System (TEKTRAN)

    We identified 14 putative full-length transcripts of proteins important for the survival of the black-winged sharpshooter, BWSS, Oncometopia nigricans. The BWSS is considered a highly competent vector of several strains of the xylem-inhabiting bacterium Xylella fastidiosa, the causal agent of a numb...

  3. Changes in the epidemiology of Pierce’s disease in California due to introduction of the glassy-winged sharpshooter

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Pierce’s disease is caused by the xylem limited bacterial pathogen, Xylella fastidiosa. This pathogen is vectored by xylem feeding insects and has been present in California for more than 100 years. Within California there are three management regions that have different key vectors: the North Coa...

  4. The Effect of Irrigation on the Temporal Increase of Citrus Variegated Chlorosis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The incidence and severity of citrus variegated chlorosis (CVC) caused by Xylella fastidiosa is higher in the northern region than in the southern region of Sao Paulo, Brazil. This is in part due to differences in the climate and especially rainfall, as the northern region tends to be drier than th...

  5. HYDRAULIC DISRUPTION AND PASSIVE MIGRATION BY A BACTERIAL PATHOGEN IN OAK TREE XYLEM

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Xylella fastidiosa (Xf) is a xylem-limited bacterial pathogen that causes leaf scorch symptoms in numerous host plant species in urban, agricultural, and natural ecosystems worldwide. The exact mechanism of hydraulic disruption and systemic colonization of xylem by Xf remains elusive across all hos...

  6. Population genetics of Homalodisca vitripennis reovirus validates timing and limited introduction to California of its invasive insect host, the glassy-winged sharpshooter

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The glassy-winged sharpshooter (GWSS; Homalodisca vitripennis Germar) is an invasive insect introduced to California circa 1989. Native to the southeastern U.S. and northeastern Mexico, GWSS is of economic concern as a vector of the Pierce’s disease bacterium Xylella fastidiosa. Recently, a novel ...

  7. Differential reproductive maturity between allopatric populations of Homalodisca vitripennis (Hemiptera: Cicadellidae) in California

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Homalodisca vitripennis (Germar) is native to southeastern U.S. and northeastern Mexico. It was detected in southern California in the late 1980s and in the San Joaquin Valley in 1999, where it transmits the bacterium Xylella fastidiosa Wells et al. to grapevines and other crops. The reproductive ...

  8. Cells derived from the glassy-winged sharpshooter, Homalodisca vitripennis, support infection and replication of viral RNA from a clone of Homalodisca coagulata virus 1 (HoCV-1).

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Pierce’s disease is a common and devastating disease of grapevines caused by the bacterium Xylella fastidiosa. The bacterium is often transmitted to plants by the glassy-winged sharpshooter (GWSS), Homalodisca vitripennis (Hemiptera: Cicadellidae) which has become a high profile pest insect targete...

  9. Congener response reduces risks from bottom-up and top-down forces: Behavioral parsimony by Xylophage

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Organisms must find food and avoid enemies. Homalodisca vitripennis (Germar), the glassy-winged sharpshooter leafhopper, is a primary vector of Xylella fastidiosa, the causal agent of Pierce's disease, a deadly disease of grapevines, fruit trees, and other woody plants. H. vitripennis feeds on xylem...

  10. Optimizing EPG settings to record blue-green sharpshooter X waves for future studies of grape host plant resistance to Xf inoculation

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The long-term goal of the research reported in this review is to develop methodology for assessment of grapevine resistant to sharpshooter inoculation of Xylella fastidiosa(Xf)into healthy grapevines, thereby preventing Xf infection. Such a trait would be quite different from the more common mechani...

  11. EM International. Volume 1

    SciTech Connect

    Not Available

    1993-07-01

    It is the intent of EM International to describe the Office of Environmental Restoration and Waste Management`s (EM`s) various roles and responsibilities within the international community. Cooperative agreements and programs, descriptions of projects and technologies, and synopses of visits to international sites are all highlighted in this semiannual journal. Focus on EM programs in this issue is on international collaboration in vitrification projects. Technology highlights covers: in situ sealing for contaminated sites; and remote sensors for toxic pollutants. Section on profiles of countries includes: Arctic contamination by the former Soviet Union, and EM activities with Germany--cooperative arrangements.

  12. Rapid, specific and quantitative assays for the detection of the endophytic bacterium Methylobacterium mesophilicum in plants.

    PubMed

    Lacava, P T; Li, W B; Araújo, W L; Azevedo, J L; Hartung, J S

    2006-06-01

    Xylella fastidiosa is a xylem-limited bacterium that causes citrus variegated chlorosis disease in sweet orange. There is evidence that X. fastidiosa interacts with endophytic bacteria present in the xylem of sweet orange, and that these interactions, particularly with Methylobacterium mesophilicum, may affect disease progress. However, these interactions cannot be evaluated in detail until efficient methods for detection and enumeration of these bacteria in planta are developed. We have previously developed standard and quantitative PCR-based assays specific for X. fastidiosa using the LightCycler system [Li, W.B., Pria Jr., L.P.M.W.D., X. Qin, and J.S. Hartung, 2003. Presence of Xylella fastidiosa in sweet orange fruit and seeds and its transmission to seedlings. Phytopathology 93:953-958.], and now report the development of both standard and quantitative PCR assays for M. mesophilicum. The assays are specific for M. mesophilicum and do not amplify DNA from other species of Methylobacterium or other bacteria commonly associated with citrus or plant tissue. Other bacteria tested included Curtobacterium flaccumfaciens, Pantoea agglomerans, Enterobacter cloacae, Bacillus sp., X. fastidiosa, Xanthomonas axonopodis pv. citri, and Candidatus Liberibacter asiaticus. We have demonstrated that with these methods we can quantitatively monitor the colonization of xylem by M. mesophilicum during the course of disease development in plants artificially inoculated with both bacteria. PMID:16266765

  13. Control of Pierce's Disease by Phage

    PubMed Central

    Das, Mayukh; Bhowmick, Tushar Suvra; Ahern, Stephen J.; Young, Ry; Gonzalez, Carlos F.

    2015-01-01

    Pierce’s Disease (PD) of grapevines, caused by Xylella fastidiosa subsp. fastidiosa (Xf), is a limiting factor in the cultivation of grapevines in the US. There are presently no effective control methods to prevent or treat PD. The therapeutic and prophylactic efficacy of a phage cocktail composed of four virulent (lytic) phages was evaluated for control of PD. Xf levels in grapevines were significantly reduced in therapeutically or prophylactically treated grapevines. PD symptoms ceased to progress one week post-therapeutic treatment and symptoms were not observed in prophylactically treated grapevines. Cocktail phage levels increased in grapevines in the presence of the host. No in planta phage-resistant Xf isolates were obtained. Moreover, Xf mutants selected for phage resistance in vitro did not cause PD symptoms. Our results indicate that phages have great potential for biocontrol of PD and other economically important diseases caused by Xylella. PMID:26107261

  14. Control of Pierce's Disease by Phage.

    PubMed

    Das, Mayukh; Bhowmick, Tushar Suvra; Ahern, Stephen J; Young, Ry; Gonzalez, Carlos F

    2015-01-01

    Pierce's Disease (PD) of grapevines, caused by Xylella fastidiosa subsp. fastidiosa (Xf), is a limiting factor in the cultivation of grapevines in the US. There are presently no effective control methods to prevent or treat PD. The therapeutic and prophylactic efficacy of a phage cocktail composed of four virulent (lytic) phages was evaluated for control of PD. Xf levels in grapevines were significantly reduced in therapeutically or prophylactically treated grapevines. PD symptoms ceased to progress one week post-therapeutic treatment and symptoms were not observed in prophylactically treated grapevines. Cocktail phage levels increased in grapevines in the presence of the host. No in planta phage-resistant Xf isolates were obtained. Moreover, Xf mutants selected for phage resistance in vitro did not cause PD symptoms. Our results indicate that phages have great potential for biocontrol of PD and other economically important diseases caused by Xylella. PMID:26107261

  15. Genome Sequence of Streptomyces wadayamensis Strain A23, an Endophytic Actinobacterium from Citrus reticulata.

    PubMed

    de Oliveira, Luciana G; Tormet Gonzalez, Gabriela D; Samborsky, Markyian; Marcon, Joelma; Araujo, Welington L; de Azevedo, João Lucio

    2014-01-01

    The actinobacterium Streptomyces wadayamensis A23 is an endophyte of Citrus reticulata that produces the antimycin and mannopeptimycin antibiotics, among others. The strain has the capability to inhibit Xylella fastidiosa growth. The draft genome of S. wadayamensis A23 has ~7.0 Mb and 6,006 protein-coding sequences, with a 73.5% G+C content. PMID:24994795

  16. emGain: Determination of EM gain of CCD

    NASA Astrophysics Data System (ADS)

    Daigle, Olivier; Carignan, Claude; Blais-Ouellette, Sebastien

    2012-01-01

    The determination of the EM gain of the CCD is best done by fitting the histogram of many low-light frames. Typically, the dark+CIC noise of a 30ms frame itself is a sufficient amount of signal to determine accurately the EM gain with about 200 512x512 frames. The IDL code emGain takes as an input a cube of frames and fit the histogram of all the pixels with the EM stage output probability function. The function returns the EM gain of the frames as well as the read-out noise and the mean signal level of the frames.

  17. The EM Earthquake Precursor

    NASA Astrophysics Data System (ADS)

    Jones, K. B., II; Saxton, P. T.

    2013-12-01

    Many attempts have been made to determine a sound forecasting method regarding earthquakes and warn the public in turn. Presently, the animal kingdom leads the precursor list alluding to a transmission related source. By applying the animal-based model to an electromagnetic (EM) wave model, various hypotheses were formed, but the most interesting one required the use of a magnetometer with a differing design and geometry. To date, numerous, high-end magnetometers have been in use in close proximity to fault zones for potential earthquake forecasting; however, something is still amiss. The problem still resides with what exactly is forecastable and the investigating direction of EM. After the 1989 Loma Prieta Earthquake, American earthquake investigators predetermined magnetometer use and a minimum earthquake magnitude necessary for EM detection. This action was set in motion, due to the extensive damage incurred and public outrage concerning earthquake forecasting; however, the magnetometers employed, grounded or buried, are completely subject to static and electric fields and have yet to correlate to an identifiable precursor. Secondly, there is neither a networked array for finding any epicentral locations, nor have there been any attempts to find even one. This methodology needs dismissal, because it is overly complicated, subject to continuous change, and provides no response time. As for the minimum magnitude threshold, which was set at M5, this is simply higher than what modern technological advances have gained. Detection can now be achieved at approximately M1, which greatly improves forecasting chances. A propagating precursor has now been detected in both the field and laboratory. Field antenna testing conducted outside the NE Texas town of Timpson in February, 2013, detected three strong EM sources along with numerous weaker signals. The antenna had mobility, and observations were noted for recurrence, duration, and frequency response. Next, two

  18. Bioterrorism awareness for EMS.

    PubMed

    Patrick, Richard W

    2004-04-01

    It is important to understand that the issues surrounding bioterrorism and all weapons of mass destruction are complex. In an effort to enhance response to such events, EMS should handle all incidents from the perspective of an all-hazards approach. Prevention, preparation, response and recovery are essential to the safe mitigation of all incidents. Organizations must be prepared. Plan now for a safer tomorrow. Your personnel and communities depend on you. PMID:15131906

  19. Anterior Foregut Microbiota of the Glassy-Winged Sharpshooter Explored Using Deep 16S rRNA Gene Sequencing from Individual Insects

    PubMed Central

    Rogers, Elizabeth E.; Backus, Elaine A.

    2014-01-01

    The glassy-winged sharpshooter (GWSS) is an invasive insect species that transmits Xylella fastidiosa, the bacterium causing Pierce's disease of grapevine and other leaf scorch diseases. X. fastidiosa has been shown to colonize the anterior foregut (cibarium and precibarium) of sharpshooters, where it may interact with other naturally-occurring bacterial species. To evaluate such interactions, a comprehensive list of bacterial species associated with the sharpshooter cibarium and precibarium is needed. Here, a survey of microbiota associated with the GWSS anterior foregut was conducted. Ninety-six individual GWSS, 24 from each of 4 locations (Bakersfield, CA; Ojai, CA; Quincy, FL; and a laboratory colony), were characterized for bacteria in dissected sharpshooter cibaria and precibaria by amplification and sequencing of a portion of the 16S rRNA gene using Illumina MiSeq technology. An average of approximately 150,000 sequence reads were obtained per insect. The most common genus detected was Wolbachia; sequencing of the Wolbachia ftsZ gene placed this strain in supergroup B, one of two Wolbachia supergroups most commonly associated with arthropods. X. fastidiosa was detected in all 96 individuals examined. By multilocus sequence typing, both X. fastidiosa subspecies fastidiosa and subspecies sandyi were present in GWSS from California and the colony; only subspecies fastidiosa was detected in GWSS from Florida. In addition to Wolbachia and X. fastidiosa, 23 other bacterial genera were detected at or above an average incidence of 0.1%; these included plant-associated microbes (Methylobacterium, Sphingomonas, Agrobacterium, and Ralstonia) and soil- or water-associated microbes (Anoxybacillus, Novosphingobium, Caulobacter, and Luteimonas). Sequences belonging to species of the family Enterobacteriaceae also were detected but it was not possible to assign these to individual genera. Many of these species likely interact with X. fastidiosa in the cibarium and

  20. Anterior foregut microbiota of the glassy-winged sharpshooter explored using deep 16S rRNA gene sequencing from individual insects.

    PubMed

    Rogers, Elizabeth E; Backus, Elaine A

    2014-01-01

    The glassy-winged sharpshooter (GWSS) is an invasive insect species that transmits Xylella fastidiosa, the bacterium causing Pierce's disease of grapevine and other leaf scorch diseases. X. fastidiosa has been shown to colonize the anterior foregut (cibarium and precibarium) of sharpshooters, where it may interact with other naturally-occurring bacterial species. To evaluate such interactions, a comprehensive list of bacterial species associated with the sharpshooter cibarium and precibarium is needed. Here, a survey of microbiota associated with the GWSS anterior foregut was conducted. Ninety-six individual GWSS, 24 from each of 4 locations (Bakersfield, CA; Ojai, CA; Quincy, FL; and a laboratory colony), were characterized for bacteria in dissected sharpshooter cibaria and precibaria by amplification and sequencing of a portion of the 16S rRNA gene using Illumina MiSeq technology. An average of approximately 150,000 sequence reads were obtained per insect. The most common genus detected was Wolbachia; sequencing of the Wolbachia ftsZ gene placed this strain in supergroup B, one of two Wolbachia supergroups most commonly associated with arthropods. X. fastidiosa was detected in all 96 individuals examined. By multilocus sequence typing, both X. fastidiosa subspecies fastidiosa and subspecies sandyi were present in GWSS from California and the colony; only subspecies fastidiosa was detected in GWSS from Florida. In addition to Wolbachia and X. fastidiosa, 23 other bacterial genera were detected at or above an average incidence of 0.1%; these included plant-associated microbes (Methylobacterium, Sphingomonas, Agrobacterium, and Ralstonia) and soil- or water-associated microbes (Anoxybacillus, Novosphingobium, Caulobacter, and Luteimonas). Sequences belonging to species of the family Enterobacteriaceae also were detected but it was not possible to assign these to individual genera. Many of these species likely interact with X. fastidiosa in the cibarium and

  1. Identified EM Earthquake Precursors

    NASA Astrophysics Data System (ADS)

    Jones, Kenneth, II; Saxton, Patrick

    2014-05-01

    Many attempts have been made to determine a sound forecasting method regarding earthquakes and warn the public in turn. Presently, the animal kingdom leads the precursor list alluding to a transmission related source. By applying the animal-based model to an electromagnetic (EM) wave model, various hypotheses were formed, but the most interesting one required the use of a magnetometer with a differing design and geometry. To date, numerous, high-end magnetometers have been in use in close proximity to fault zones for potential earthquake forecasting; however, something is still amiss. The problem still resides with what exactly is forecastable and the investigating direction of EM. After a number of custom rock experiments, two hypotheses were formed which could answer the EM wave model. The first hypothesis concerned a sufficient and continuous electron movement either by surface or penetrative flow, and the second regarded a novel approach to radio transmission. Electron flow along fracture surfaces was determined to be inadequate in creating strong EM fields, because rock has a very high electrical resistance making it a high quality insulator. Penetrative flow could not be corroborated as well, because it was discovered that rock was absorbing and confining electrons to a very thin skin depth. Radio wave transmission and detection worked with every single test administered. This hypothesis was reviewed for propagating, long-wave generation with sufficient amplitude, and the capability of penetrating solid rock. Additionally, fracture spaces, either air or ion-filled, can facilitate this concept from great depths and allow for surficial detection. A few propagating precursor signals have been detected in the field occurring with associated phases using custom-built loop antennae. Field testing was conducted in Southern California from 2006-2011, and outside the NE Texas town of Timpson in February, 2013. The antennae have mobility and observations were noted for

  2. Cicadomorpha insects associated with bacterial leaf scorch infected oak in central New Jersey.

    PubMed

    Zhang, Jianxin; Lashomb, James; Gould, Ann; Hamilton, George

    2011-10-01

    Potential insect vectors for transmission of oak leaf scorch caused by Xylella fastidiosa Wells et al., in pin and red oaks in New Jersey were surveyed by placing yellow sticky card traps in tree canopies and fogging with pyrethrin insecticide during 2002-2006. Thirty-seven Cicadomorpha species were collected from 20 genera in Membracidae, Cicadellidae, Aphrophoridae, and Clastopteridae. Of the 12,880 potential vectors collected, 91.4% were Membracidae, 6.9% were Cicadellidae, and 1.7% were Aphrophoridae and Clastopteridae. Fogging collected more insect species and individuals than sticky card collections. Sticky card sampling, done more frequently and at a larger number of locations provided similar community structure information as fogging. Sticky card collections of the dominant treehopper species, Ophiderma definita Woodruff were male biased when females were gravid. O. definita populations peaked in early June, comprised 68.2% of the total collection, and were more abundant in pin oaks than red oaks. Graphocephala versuta (Say) peaked in mid-July, comprising 6.2% of the total collection. Higher Cicadomorpha populations were observed in asymptomatic oak canopies than in neighboring X. fastidiosa infected oaks. Individual insect specimens collected from oaks were subjected to a X. fastidiosa DNA assay by polymerase chain reaction amplification. The average X. fastidiosa positive rate was 13.89% for all specimens tested. Eleven treehopper species, six leafhopper species, and four spittlebug species tested DNA positive for X. fastidiosa. PMID:22251725

  3. Disrupting the Transmission of a Vector-Borne Plant Pathogen

    PubMed Central

    Rashed, Arash; Almeida, Rodrigo P. P.

    2012-01-01

    Approaches to control vector-borne diseases rarely focus on the interface between vector and microbial pathogen, but strategies aimed at disrupting the interactions required for transmission may lead to reductions in disease spread. We tested if the vector transmission of the plant-pathogenic bacterium Xylella fastidiosa was affected by three groups of molecules: lectins, carbohydrates, and antibodies. Although not comprehensively characterized, it is known that X. fastidiosa adhesins bind to carbohydrates, and that these interactions are important for initial cell attachment to vectors, which is required for bacterial transmission from host to host. Lectins with affinity to substrates expected to occur on the cuticular surface of vectors colonized by X. fastidiosa, such as wheat germ agglutinin, resulted in statistically significant reductions in transmission rate, as did carbohydrates with N-acetylglucosamine residues. Presumably, lectins bound to receptors on the vector required for cell adhesion/colonization, while carbohydrate-saturated adhesins on X. fastidiosa's cell surface. Furthermore, antibodies against X. fastidiosa whole cells, gum, and afimbrial adhesins also resulted in transmission blockage. However, no treatment resulted in the complete abolishment of transmission, suggesting that this is a complex biological process. This work illustrates the potential to block the transmission of vector-borne pathogens without directly affecting either organism. PMID:22101059

  4. Disrupting the transmission of a vector-borne plant pathogen.

    PubMed

    Killiny, Nabil; Rashed, Arash; Almeida, Rodrigo P P

    2012-02-01

    Approaches to control vector-borne diseases rarely focus on the interface between vector and microbial pathogen, but strategies aimed at disrupting the interactions required for transmission may lead to reductions in disease spread. We tested if the vector transmission of the plant-pathogenic bacterium Xylella fastidiosa was affected by three groups of molecules: lectins, carbohydrates, and antibodies. Although not comprehensively characterized, it is known that X. fastidiosa adhesins bind to carbohydrates, and that these interactions are important for initial cell attachment to vectors, which is required for bacterial transmission from host to host. Lectins with affinity to substrates expected to occur on the cuticular surface of vectors colonized by X. fastidiosa, such as wheat germ agglutinin, resulted in statistically significant reductions in transmission rate, as did carbohydrates with N-acetylglucosamine residues. Presumably, lectins bound to receptors on the vector required for cell adhesion/colonization, while carbohydrate-saturated adhesins on X. fastidiosa's cell surface. Furthermore, antibodies against X. fastidiosa whole cells, gum, and afimbrial adhesins also resulted in transmission blockage. However, no treatment resulted in the complete abolishment of transmission, suggesting that this is a complex biological process. This work illustrates the potential to block the transmission of vector-borne pathogens without directly affecting either organism. PMID:22101059

  5. EPA LABORATORIES IMPLEMENT EMS PROGRAM

    EPA Science Inventory

    This paper highlights the breadth and magnitude of carrying out an effective Environmental Management System (EMS) program at the U.S. EPA's research and development laboratories. Federal research laboratories have unique operating challenges compared to more centralized industr...

  6. Thiazomycins, thiazolyl peptide antibiotics from Amycolatopsis fastidiosa.

    PubMed

    Zhang, Chaowei; Herath, Kithsiri; Jayasuriya, Hiranthi; Ondeyka, John G; Zink, Deborah L; Occi, James; Birdsall, Gwyneth; Venugopal, Jayashree; Ushio, Misti; Burgess, Bruce; Masurekar, Prakash; Barrett, John F; Singh, Sheo B

    2009-05-22

    Thiazolyl peptides are a class of highly rigid trimacrocyclic compounds consisting of varying but large numbers of thiazole rings. The need for new antibacterial agents to treat infections caused by resistant bacteria prompted a reinvestigation of this class, leading to the previous isolation of thiazolyl peptides, namely, thiazomycin (5) and thiazomycin A (6), congeners of nocathiacins (1-4). Continued chemical screening led to the isolation of six new thiazolyl peptide congeners (8-13), of which three had truncated structures lacking an indole residue. From these, compound 8 showed activity similar to thiazomycin. Two compounds (9 and 10) showed intermediate activities, and the three truncated compounds (11-13) were essentially inactive. The discovery of the truncated compounds revealed the minimal structural requirements for activity and suggested probable biosynthetic pathways for more advanced compounds. The isolation, structure elucidation, antibacterial activity, and proposed biogenesis of thiazomycins are herein described. PMID:19334707

  7. Is EMS communicating with the FCC?

    PubMed

    Johnson, M S; VanCott, C; Glass, C; Anderson, P B

    1989-07-01

    Radio communication problems in EMS run the spectrum from annoying to deadly. Dedicated radio frequencies for EMS, much like those exclusive to police and fire departments, are long overdue. PMID:10293680

  8. Alveolar Echinococcosis: Characterization of Diagnostic Antigen Em18 and Serological Evaluation of Recombinant Em18

    PubMed Central

    Sako, Yasuhito; Nakao, Minoru; Nakaya, Kazuhiro; Yamasaki, Hiroshi; Gottstein, Bruno; Lightowers, Marshall W.; Schantz, Peter M.; Ito, Akira

    2002-01-01

    The Echinococcus multilocularis protein Em18 is one of the most promising antigens for use in serodiagnosis of alveolar echinococcosis in human patients. Here we identify an antigenic relationship between Em18 and a 65-kDa immunodominant E. multilocularis surface protein previously identified as either EM10 or EmII/3. The NH2-terminal sequence of native Em18 was determined, revealing it to be a fragment of EM10. Experiments were undertaken to investigate the effect of proteinase inhibitors on the degradation of EM10 in crude extracts of E. multilocularis protoscoleces. Em18 was found to be the product of degradation of EM10 by cysteine proteinase. A recombinant Em18 (RecEm18, derived from 349K to 508K of EM10) was successfully expressed by using Escherichia coli expression system and then evaluated for use in serodiagnosis of alveolar echinococcosis. RecEm18 was recognized by 27 (87.1%) and 28 (90.3%) of 31 serum samples from clinically and/or pathologically confirmed alveolar echinococcosis patients by enzyme-linked immunosorbent assay and immunoblotting, respectively. Of 33 serum samples from cystic echinococcosis patients, 1 was recorded as having a weak positive reaction to RecEm18; however, none of the serum samples which were tested from neurocysticercosis patients (n = 10) or healthy people (n = 15) showed positive reactions. RecEm18 has the potential for use in the differential serodiagnosis of alveolar echinococcosis. PMID:12149326

  9. The European Mobile System (EMS)

    NASA Technical Reports Server (NTRS)

    Jongejans, A.; Rogard, R.; Mistretta, I.; Ananasso, F.

    1993-01-01

    The European Space Agency is presently procuring an L band payload in order to promote a regional European L band system coping with the specific needs of the European market. The payload, and the two communications systems to be supported, are described below. The potential market for EMS in Europe is discussed.

  10. A Two-Dimensional Multiphase Model of Biofilm Formation in Microfluidic Chambers.

    PubMed

    Whidden, Mark; Cogan, Nick; Donahue, Matt; Navarrete, Fernando; De La Fuente, Leonardo

    2015-12-01

    The bacterial pathogen Xylella fastidiosa is the causal agent of many pathological conditions of economically important agricultural crops. There is no known cure for X. fastidiosa diseases, and management of the problem is based solely in controlling the population of insect vectors, which is somewhat effective. The bacterium causes disease by forming biofilms inside the vascular system of the plant, a process that is poorly understood. In microfluidic chambers, used as artificial xylem vessels, this bacterium has been observed to reproducibly cluster into a distinct, regular pattern of aggregates, spatially separated by channels of non-biofilm components. We develop a multiphase model in two dimensions, which recapitulates this spatial patterning, suggesting that bacterial growth and attachment/detachment processes are strongly influential modulators of these patterns. This indicates plausible strategies, such as the addition of metals and chelators, for mitigating the severity of diseases induced by this bacterial pathogen. PMID:26621357

  11. Compositions and Methods for the Treatment of Pierce's Disease

    DOEpatents

    Gupta, Goutam

    2008-10-07

    Chimeric anti-microbial proteins, compositions, and methods for the therapeutic and prophylactic treatment of plant diseases caused by the bacterial pathogen Xylella fastidiosa are provided. The anti-microbial proteins of the invention generally comprise a surface recognition domain polypeptide, capable of binding to a bacterial membrane component, fused to a bacterial lysis domain polypeptide, capable of affecting lysis or rupture of the bacterial membrane, typically via a fused polypeptide linker. In particular, methods and compositions for the treatment or prevention of Pierce's disease of grapevines are provided. Methods for the generation of transgenic Vitus vinefera plants expressing xylem-secreted anti-microbial chimeras are also provided.

  12. Bacteria causing important diseases of citrus utilise distinct modes of pathogenesis to attack a common host.

    PubMed

    Vojnov, Adrián Alberto; do Amaral, Alexandre Morais; Dow, John Maxwell; Castagnaro, Atilio Pedro; Marano, Marìa Rosa

    2010-06-01

    In this review, we summarise the current knowledge on three pathogens that exhibit distinct tissue specificity and modes of pathogenesis in citrus plants. Xanthomonas axonopodis pv. citri causes canker disease and invades the host leaf mesophyll tissue through natural openings and can also survive as an epiphyte. Xylella fastidiosa and Candidatus Liberibacter are vectored by insects and proliferate in the vascular system of the host, either in the phloem (Candidatus Liberibacter) or xylem (X. fastidiosa) causing variegated chlorosis and huanglongbing diseases, respectively. Candidatus Liberibacter can be found within host cells and is thus unique as an intracellular phytopathogenic bacterium. Genome sequence comparisons have identified groups of species-specific genes that may be associated with the particular lifestyle, mode of transmission or symptoms produced by each phytopathogen. In addition, components that are conserved amongst bacteria may have diverse regulatory actions underpinning the different bacterial lifestyles; one example is the divergent role of the Rpf/DSF cell-cell signalling system in X. citri and X. fastidiosa. Biofilm plays a key role in epiphytic fitness and canker development in X. citri and in the symptoms produced by X. fastidiosa. Bacterial aggregation may be associated with vascular occlusion of the xylem vessels and symptomatology of variegated chlorosis. PMID:20449739

  13. DOE/EM Criticality Safety Needs Assessment

    SciTech Connect

    Westfall, Robert Michael; Hopper, Calvin Mitchell

    2011-02-01

    The issue of nuclear criticality safety (NCS) in Department of Energy Environmental Management (DOE/EM) fissionable material operations presents challenges because of the large quantities of material present in the facilities and equipment that are committed to storage and/or material conditioning and dispositioning processes. Given the uncertainty associated with the material and conditions for many DOE/EM fissionable material operations, ensuring safety while maintaining operational efficiency requires the application of the most-effective criticality safety practices. In turn, more-efficient implementation of these practices can be achieved if the best NCS technologies are utilized. In 2002, DOE/EM-1 commissioned a survey of criticality safety technical needs at the major EM sites. These needs were documented in the report Analysis of Nuclear Criticality Safety Technology Supporting the Environmental Management Program, issued May 2002. Subsequent to this study, EM safety management personnel made a commitment to applying the best and latest criticality safety technology, as described by the DOE Nuclear Criticality Safety Program (NCSP). Over the past 7 years, this commitment has enabled the transfer of several new technologies to EM operations. In 2008, it was decided to broaden the basis of the EM NCS needs assessment to include not only current needs for technologies but also NCS operational areas with potential for improvements in controls, analysis, and regulations. A series of NCS workshops has been conducted over the past years, and needs have been identified and addressed by EM staff and contractor personnel. These workshops were organized and conducted by the EM Criticality Safety Program Manager with administrative and technical support by staff at Oak Ridge National Laboratory (ORNL). This report records the progress made in identifying the needs, determining the approaches for addressing these needs, and assimilating new NCS technologies into EM

  14. School Budget Hold'em Facilitator's Guide

    ERIC Educational Resources Information Center

    Education Resource Strategies, 2012

    2012-01-01

    "School Budget Hold'em" is a game designed to help school districts rethink their budgeting process. It evolved out of Education Resource Strategies' (ERS) experience working with large urban districts around the country. "School Budget Hold'em" offers a completely new approach--one that can turn the budgeting process into a long-term visioning…

  15. View of Spacelab engineering Model (EM)

    NASA Technical Reports Server (NTRS)

    1981-01-01

    View of Spacelab engineering Model (EM) as it is being brought in the O and C bldg at Kenndey Space Center (27464); view of the EM as it is being offloaded from the C-54 aircraft. Kennedy Space Center alternative photo number is 108-KSC-80-OC-666 (27465); model taken out to launch pad (27466).

  16. EM international activities. February 1997 highlights

    SciTech Connect

    1997-02-01

    EM International Highlights is a brief summary of on-going international projects within the Department of Energy`s Office of Environmental Management (EM). This document contains sections on: Global Issues, activities in Western Europe, activities in central and Eastern Europe, activities in Russia, activities in Asia and the Pacific Rim, activities in South America, activities in North America, and International Organizations.

  17. EM International, July 1994, Volume 2

    SciTech Connect

    Not Available

    1994-10-01

    The Office of Environmental Management (EM) at the Department of Energy (DOE) is seeking out and leveraging foreign technology, data, and resources in keeping with EM`s mandate to protect public health and the environment through the safe and cost-effective remediation of the Department`s nuclear weapons sites. EM works closely with foreign governments, industry, and universities to obtain innovative environmental technologies, scientific and engineering expertise, and operations experience that will support EM`s objectives. Where appropriate, these international resources are used to manage the more urgent risks at our sites, secure a safe workplace, help build consensus on critical issues, and strengthen our technology development program. Through international agreements EM engages in cooperative exchange of information, technology, and individuals. Currently, we are managing agreements with a dozen countries in Europe, Latin America, and Asia. These agreements focus on environmental restoration, waste management, transportation of radioactive wastes, and decontamination and decommissioning. This publication contains the following articles: in situ remediation integrated program; in-situ characterization and inspection of tanks; multimedia environmental pollutant assessment system (MEPAS); LLNL wet oxidation -- AEA technology. Besides these articles, this publication covers: EU activities with Russia; technology transfer activities; and international organization activities.

  18. Many factors complicate EM susceptibility tests

    NASA Astrophysics Data System (ADS)

    Richards, R. J.

    1982-09-01

    Procedures and apparatus currently employed for assaying the EM susceptibility of communication, navigation, and EW equipment are described. Susceptibility is examined in either conducted susceptibility tests, where signals are introduced into the input port of the device under test, or in radiated modes, where the entire device is exposed to an EM field to test for component and system failure. Noting that military standards require up to 10 times the EM resistance as commercial standards, the use of shielded enclosures in both commercial and military testing facilities is explored. RF-tight enclosures are filled with a homogeneous EM field produced by, optimally, broadband generators which emit signals which are amplified to desired levels. Sweep functions permit testing under broadband conditions. Attention is given to radiator selection and antenna choice to produce satisfactory test conditions at all frequencies.

  19. Unified Data Resource for CryoEM

    PubMed Central

    Lawson, Catherine L.

    2010-01-01

    3D cryo-electron microscopy reconstruction methods are uniquely able to reveal structures of many important macromolecules and macromolecular complexes. EMDataBank.org, a joint effort of the Protein Data Bank in Europe (PDBe), the Research Collaboratory for Structural Bioinformatics (RCSB), and the National Center for Macromolecular Imaging (NCMI), is a “one-stop shop” resource for global deposition and retrieval of cryoEM map, model and associated metadata. The resource unifies public access to the two major EM Structural Data archives: EM Data Bank (EMDB) and Protein Data Bank (PDB), and facilitates use of EM structural data of macromolecules and macromolecular complexes by the wider scientific community. PMID:20888470

  20. EMS in Taiwan: past, present, and future.

    PubMed

    Chiang, Wen-Chu; Ko, Patrick Chow-In; Wang, Hui-Chih; Yang, Chi-Wei; Shih, Fuh-Yuan; Hsiung, Kuang-Hua; Ma, Matthew Huei-Ming

    2009-01-01

    Taiwan is a small island country located in East Asia. From around 1995 modern concepts of the EMS were imported and supported by legislation. Considerable progress has since been made towards the construction of an effective pre-hospital care system. This article introduces the current status of the EMS in Taiwan, including the systems, response configurations, funding, personnel, medical directorship, and outcome research. The features and problems of in-hospital emergency care are also discussed. Key areas for further development in the country vary depending on regional differences in available resource and population density. An analysis of the strength, weakness, opportunity, and threats of the evolving EMS in Taiwan could be an example for other countries where the EMS is undergoing a similar process of development and optimisation. PMID:19059690

  1. EMS adaptation for climate change

    NASA Astrophysics Data System (ADS)

    Pan, C.; Chang, Y.; Wen, J.; Tsai, M.

    2010-12-01

    The purpose of this study was to find an appropriate scenario of pre-hospital transportation of an emergency medical service (EMS) system for burdensome casualties resulting from extreme climate events. A case of natural catastrophic events in Taiwan, 88 wind-caused disasters, was reviewed and analyzed. A sequential-conveyance method was designed to shorten the casualty transportation time and to promote the efficiency of ambulance services. A proposed mobile emergency medical center was first constructed in a safe area, but nearby the disaster area. The Center consists of professional medical personnel who process the triage of incoming patients and take care of casualties with minor injuries. Ambulances in the Center were ready to sequentially convey the casualties with severer conditions to an assigned hospital that is distant from the disaster area for further treatment. The study suggests that if we could construct a spacious and well-equipped mobile emergency medical center, only a small portion of casualties would need to be transferred to distant hospitals. This would reduce the over-crowding problem in hospital ERs. First-line ambulances only reciprocated between the mobile emergency medical center and the disaster area, saving time and shortening the working distances. Second-line ambulances were highly regulated between the mobile emergency medical center and requested hospitals. The ambulance service of the sequential-conveyance method was found to be more efficient than the conventional method and was concluded to be more profitable and reasonable on paper in adapting to climate change. Therefore, additional practical work should be launched to collect more precise quantitative data.

  2. 10 CFR Appendixes E-M to Part 52 - [Reserved

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 10 Energy 2 2010-01-01 2010-01-01 false E Appendixes E-M to Part 52 Energy NUCLEAR REGULATORY COMMISSION (CONTINUED) LICENSES, CERTIFICATIONS, AND APPROVALS FOR NUCLEAR POWER PLANTS Appendixes E-M to Part 52...

  3. 10 CFR Appendixes E-M to Part 52 - [Reserved

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 10 Energy 2 2014-01-01 2014-01-01 false E Appendixes E-M to Part 52 Energy NUCLEAR REGULATORY COMMISSION (CONTINUED) LICENSES, CERTIFICATIONS, AND APPROVALS FOR NUCLEAR POWER PLANTS Appendixes E-M to Part 52...

  4. 10 CFR Appendixes E-M to Part 52 - [Reserved

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 10 Energy 2 2012-01-01 2012-01-01 false E Appendixes E-M to Part 52 Energy NUCLEAR REGULATORY COMMISSION (CONTINUED) LICENSES, CERTIFICATIONS, AND APPROVALS FOR NUCLEAR POWER PLANTS Appendixes E-M to Part 52...

  5. 10 CFR Appendixes E-M to Part 52 - [Reserved

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 10 Energy 2 2013-01-01 2013-01-01 false E Appendixes E-M to Part 52 Energy NUCLEAR REGULATORY COMMISSION (CONTINUED) LICENSES, CERTIFICATIONS, AND APPROVALS FOR NUCLEAR POWER PLANTS Appendixes E-M to Part 52...

  6. 10 CFR Appendixes E-M to Part 52 - [Reserved

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 10 Energy 2 2011-01-01 2011-01-01 false E Appendixes E-M to Part 52 Energy NUCLEAR REGULATORY COMMISSION (CONTINUED) LICENSES, CERTIFICATIONS, AND APPROVALS FOR NUCLEAR POWER PLANTS Appendixes E-M to Part 52...

  7. Processing of Cryo-EM Movie Data.

    PubMed

    Ripstein, Z A; Rubinstein, J L

    2016-01-01

    Direct detector device (DDD) cameras dramatically enhance the capabilities of electron cryomicroscopy (cryo-EM) due to their improved detective quantum efficiency (DQE) relative to other detectors. DDDs use semiconductor technology that allows micrographs to be recorded as movies rather than integrated individual exposures. Movies from DDDs improve cryo-EM in another, more surprising, way. DDD movies revealed beam-induced specimen movement as a major source of image degradation and provide a way to partially correct the problem by aligning frames or regions of frames to account for this specimen movement. In this chapter, we use a self-consistent mathematical notation to explain, compare, and contrast several of the most popular existing algorithms for computationally correcting specimen movement in DDD movies. We conclude by discussing future developments in algorithms for processing DDD movies that would extend the capabilities of cryo-EM even further. PMID:27572725

  8. Risk Communication Within the EM Program

    SciTech Connect

    Edelson, M.

    2003-02-26

    The U.S. Department of Energy Environmental Management program (EM) conducts the most extensive environmental remediation effort in the world. The annual EM budgets have exceeded $6,000,000,000 for approximately ten years and EM has assumed responsibility for the cleanup of the largest DOE reservations (i.e., at Hanford, Washington, Aiken, South Carolina, and Idaho Falls, Idaho) as well as the facilities at Rocky Flats, Colorado and in Ohio. Each of these sites has areas of extensive radioactive and chemical contamination, numerous surplus facilities that require decontamination and removal, while some have special nuclear material that requires secure storage. The EM program has been criticized for being ineffective (1) and has been repeatedly reorganized to address perceived shortcomings. The most recent reorganization was announced in 2001 to become effective at the beginning of the 2003 Federal Fiscal Year (i.e., October 2002). It was preceded by a ''top to bottom'' review (TTBR) of the program (2) that identified several deficiencies that were to be corrected as a result of the reorganization. One prominent outcome of the TTBR was the identification of ''risk reduction'' as an organizing principle to prioritize the activities of the new EM program. The new program also sought to accelerate progress by identifying a set of critical activities at each site that could be accelerated and result in more rapid site closure, with attendant risk, cost, and schedule benefits. This paper investigates how the new emphasis on risk reduction in the EM program has been communicated to EM stakeholders and regulators. It focuses on the Rocky Flats Environmental Technology Site (RFETS) as a case study and finds that there is little evidence for a new emphasis on risk reduction in EM communications with RFETS stakeholders. Discussions between DOE and RFETS stakeholders often refer to ''risk,'' but the word serves as a placeholder for other concepts. Thus ''risk'' communication

  9. Biological and genetic factors regulating natural competence in a bacterial plant pathogen.

    PubMed

    Kung, Stephanie H; Almeida, Rodrigo P P

    2014-01-01

    For naturally competent bacteria, spatially structured growth can provide an environment for enhanced horizontal gene transfer through transformation and recombination. DNA is often present in the extracellular environment, such as in the extracellular matrix of biofilms, and the lysis of a single cell can result in high local DNA concentrations. Xylella fastidiosa is a naturally competent plant pathogen that typically lives in a surface-attached state, yet previous work characterizing the competence of this organism was conducted with planktonic cells in liquid environments. Here, we show that transformation and recombination efficiencies are two to three orders of magnitude higher for cells grown on solid compared with liquid media, with maximum recombination efficiencies of about 10(-3). Cells were highly competent throughout their exponential growth phase, with no significant change in recombination efficiencies until population growth rates began to slow. Mutations in type IV pili, competency-related, and cell-cell signalling genes significantly impacted the ability of X. fastidiosa to acquire and incorporate DNA. Because X. fastidiosa is highly competent when growing in a surface-attached state, as it does within its insect vectors and host plants, recombination of naturally transformed DNA could be a significant route by which horizontal gene transfer occurs in natural environments. PMID:24149707

  10. Overexpression of antimicrobial lytic peptides protects grapevine from Pierce's disease under greenhouse but not field conditions.

    PubMed

    Li, Zhijian T; Hopkins, Donald L; Gray, Dennis J

    2015-10-01

    Pierce's disease (PD) caused by Xylella fastidiosa prevents cultivation of grapevine (Vitis vinifera) and susceptible hybrids in the southeastern United States and poses a major threat to the grape industry of California and Texas. Genetic resistance is the only proven control of X. fastidiosa. Genetic engineering offers an alternative to heretofore ineffective conventional breeding in order to transfer only PD resistance traits into elite cultivars. A synthetic gene encoding lytic peptide LIMA-A was introduced into V. vinifera and a Vitis hybrid to assess in planta inhibition of X. fastidiosa. Over 1050 independent transgenic plant lines were evaluated in the greenhouse, among which nine lines were selected and tested under naturally-inoculated field conditions. These selected plant lines in the greenhouse remain disease-free for 10 years, to date, even with multiple manual pathogen inoculations. However, all these lines in the field, including a grafted transgenic rootstock, succumbed to PD within 7 years. We conclude that in planta production of antimicrobial lytic peptides does not provide durable PD resistance to grapevine under field conditions. PMID:25894660

  11. Do earthquakes generate EM signals?

    NASA Astrophysics Data System (ADS)

    Walter, Christina; Onacha, Stephen; Malin, Peter; Shalev, Eylon; Lucas, Alan

    2010-05-01

    study areas, large swarms of earthquakes were located very close to the electromagnetic coils. This abstract focuses on the data from the Wairakei area. Preliminary data analysis has been carried out by band pass filtering and removing of the harmonics of the 50 Hz power line frequency. The initial results clearly show that electromagnetic signals accompany the seismic P and S waves (coseismic signal). Further data analysis involves the extraction of the seismoelectric signal generated at the onset of the earthquake and at interfaces from the coseismic signal and other ‘noise' sources. This processing step exhibits a major challenge in seismoelectric data processing. Unlike in other studies we measured the EM field and the seismic field at one location. Therefore the seismoelectric wave travelling at the speed of light cannot be determined as easily in the arrival times as when an array of coils is used. This makes the determination of the origin time much more difficult. Hence other processing techniques need to be explored.

  12. The E-MS Algorithm: Model Selection with Incomplete Data

    PubMed Central

    Jiang, Jiming; Nguyen, Thuan; Rao, J. Sunil

    2014-01-01

    We propose a procedure associated with the idea of the E-M algorithm for model selection in the presence of missing data. The idea extends the concept of parameters to include both the model and the parameters under the model, and thus allows the model to be part of the E-M iterations. We develop the procedure, known as the E-MS algorithm, under the assumption that the class of candidate models is finite. Some special cases of the procedure are considered, including E-MS with the generalized information criteria (GIC), and E-MS with the adaptive fence (AF; Jiang et al. 2008). We prove numerical convergence of the E-MS algorithm as well as consistency in model selection of the limiting model of the E-MS convergence, for E-MS with GIC and E-MS with AF. We study the impact on model selection of different missing data mechanisms. Furthermore, we carry out extensive simulation studies on the finite-sample performance of the E-MS with comparisons to other procedures. The methodology is also illustrated on a real data analysis involving QTL mapping for an agricultural study on barley grains. PMID:26783375

  13. Percutaneous absorption and disposition of Tinopal EMS.

    PubMed

    Black, J G; Moule, R C; Philp, J

    1977-08-01

    A cotton-substantive, anionic, fluorescent whitening agent manufactured by several suppliers under various trade names e.g. Tinopal EMS, has been synthesized in radioactive form. Intubation of detergent or aqueous solution into rats resulted in little absorption from the intestinal tract as evidenced by low radioactivity in the urine and tissues. Most of the dose was excreted rapidly in the faeces. After parenteral administration to rats, the radioactivity was rapidly excreted in the faeces with small amounts remaining in tissues and organs. There was slight evidence of retention of radioactivity in the kidneys. Very small amounts of Tinopal EMS in detergent were absorbed through rat skin, but only when concentrations greater than those normally used by the consumer, together with occlusion of the skin were employed. Small amounts were absorbed throught skin when applied in ethanol. It is concluded that the possibility of systemic toxic effects in man as a result of percutaneous absorption is remote. PMID:929616

  14. Crosshole EM in steel-cased boreholes

    SciTech Connect

    Wilt, M.; Lee, K.H.; Becker, A.; Spies, B.; Wang, B.

    1996-07-01

    The application of crosshole EM methods through steel well-casing was investigated in theoretical, laboratory and field studies. A numerical code was developed that calculates the attenuation and phase delay of an EM dipole signal propagated through a steel well casing lodged in a homogeneous medium. The code was validated with a scale model and used for sensitivity studies of casing and formation properties. Finally, field measurements were made in an oil field undergoing waterflooding. Our most important findings are that (1) crosshole surveys are feasible using a well pair with one metallic and one non-metallic casing. (2) The casing effect seems be localized within the pipe section that includes the sensor. (3) The effects of the casing can be corrected using simple means and (4) crosshole field data that are sensitive to both formation and casing were acquired in a working environment.

  15. TandEM: Titan and Enceladus mission

    USGS Publications Warehouse

    Coustenis, A.; Atreya, S.K.; Balint, T.; Brown, R.H.; Dougherty, M.K.; Ferri, F.; Fulchignoni, M.; Gautier, D.; Gowen, R.A.; Griffith, C.A.; Gurvits, L.I.; Jaumann, R.; Langevin, Y.; Leese, M.R.; Lunine, J.I.; McKay, C.P.; Moussas, X.; Muller-Wodarg, I.; Neubauer, F.; Owen, T.C.; Raulin, F.; Sittler, E.C.; Sohl, F.; Sotin, C.; Tobie, G.; Tokano, T.; Turtle, E.P.; Wahlund, J.-E.; Waite, J.H.; Baines, K.H.; Blamont, J.; Coates, A.J.; Dandouras, I.; Krimigis, T.; Lellouch, E.; Lorenz, R.D.; Morse, A.; Porco, C.C.; Hirtzig, M.; Saur, J.; Spilker, T.; Zarnecki, J.C.; Choi, E.; Achilleos, N.; Amils, R.; Annan, P.; Atkinson, D.H.; Benilan, Y.; Bertucci, C.; Bezard, B.; Bjoraker, G.L.; Blanc, M.; Boireau, L.; Bouman, J.; Cabane, M.; Capria, M.T.; Chassefiere, E.; Coll, P.; Combes, M.; Cooper, J.F.; Coradini, A.; Crary, F.; Cravens, T.; Daglis, I.A.; de Angelis, E.; De Bergh, C.; de Pater, I.; Dunford, C.; Durry, G.; Dutuit, O.; Fairbrother, D.; Flasar, F.M.; Fortes, A.D.; Frampton, R.; Fujimoto, M.; Galand, M.; Grasset, O.; Grott, M.; Haltigin, T.; Herique, A.; Hersant, F.; Hussmann, H.; Ip, W.; Johnson, R.; Kallio, E.; Kempf, S.; Knapmeyer, M.; Kofman, W.; Koop, R.; Kostiuk, T.; Krupp, N.; Kuppers, M.; Lammer, H.; Lara, L.-M.; Lavvas, P.; Le, Mouelic S.; Lebonnois, S.; Ledvina, S.; Li, J.; Livengood, T.A.; Lopes, R.M.; Lopez-Moreno, J. -J.; Luz, D.; Mahaffy, P.R.; Mall, U.; Martinez-Frias, J.; Marty, B.; McCord, T.; Salvan, C.M.; Milillo, A.; Mitchell, D.G.; Modolo, R.; Mousis, O.; Nakamura, M.; Neish, C.D.; Nixon, C.A.; Mvondo, D.N.; Orton, G.; Paetzold, M.; Pitman, J.; Pogrebenko, S.; Pollard, W.; Prieto-Ballesteros, O.; Rannou, P.; Reh, K.; Richter, L.; Robb, F.T.; Rodrigo, R.; Rodriguez, S.; Romani, P.; Bermejo, M.R.; Sarris, E.T.; Schenk, P.; Schmitt, B.; Schmitz, N.; Schulze-Makuch, D.; Schwingenschuh, K.; Selig, A.; Sicardy, B.; Soderblom, L.; Spilker, L.J.; Stam, D.; Steele, A.; Stephan, K.; Strobel, D.F.; Szego, K.; Szopa

    2009-01-01

    TandEM was proposed as an L-class (large) mission in response to ESA's Cosmic Vision 2015-2025 Call, and accepted for further studies, with the goal of exploring Titan and Enceladus. The mission concept is to perform in situ investigations of two worlds tied together by location and properties, whose remarkable natures have been partly revealed by the ongoing Cassini-Huygens mission. These bodies still hold mysteries requiring a complete exploration using a variety of vehicles and instruments. TandEM is an ambitious mission because its targets are two of the most exciting and challenging bodies in the Solar System. It is designed to build on but exceed the scientific and technological accomplishments of the Cassini-Huygens mission, exploring Titan and Enceladus in ways that are not currently possible (full close-up and in situ coverage over long periods of time). In the current mission architecture, TandEM proposes to deliver two medium-sized spacecraft to the Saturnian system. One spacecraft would be an orbiter with a large host of instruments which would perform several Enceladus flybys and deliver penetrators to its surface before going into a dedicated orbit around Titan alone, while the other spacecraft would carry the Titan in situ investigation components, i.e. a hot-air balloon (Montgolfi??re) and possibly several landing probes to be delivered through the atmosphere. ?? Springer Science + Business Media B.V. 2008.

  16. Helicopter EMS: Research Endpoints and Potential Benefits

    PubMed Central

    Thomas, Stephen H.; Arthur, Annette O.

    2012-01-01

    Patients, EMS systems, and healthcare regions benefit from Helicopter EMS (HEMS) utilization. This article discusses these benefits in terms of specific endpoints utilized in research projects. The endpoint of interest, be it primary, secondary, or surrogate, is important to understand in the deployment of HEMS resources or in planning further HEMS outcomes research. The most important outcomes are those which show potential benefits to the patients, such as functional survival, pain relief, and earlier ALS care. Case reports are also important “outcomes” publications. The benefits of HEMS in the rural setting is the ability to provide timely access to Level I or Level II trauma centers and in nontrauma, interfacility transport of cardiac, stroke, and even sepsis patients. Many HEMS crews have pharmacologic and procedural capabilities that bring a different level of care to a trauma scene or small referring hospital, especially in the rural setting. Regional healthcare and EMS system's benefit from HEMS by their capability to extend the advanced level of care throughout a region, provide a “backup” for areas with limited ALS coverage, minimize transport times, make available direct transport to specialized centers, and offer flexibility of transport in overloaded hospital systems. PMID:22203905

  17. Blocking the Transmission of a Noncirculative Vector-Borne Plant Pathogenic Bacterium.

    PubMed

    Labroussaa, Fabien; Zeilinger, Adam R; Almeida, Rodrigo P P

    2016-07-01

    The successful control of insect-borne plant pathogens is often difficult to achieve due to the ecologically complex interactions among pathogens, vectors, and host plants. Disease management often relies on pesticides and other approaches that have limited long-term sustainability. To add a new tool to control vector-borne diseases, we attempted to block the transmission of a bacterial insect-transmitted pathogen, the bacterium Xylella fastidiosa, by disrupting bacteria-insect vector interactions. X. fastidiosa is known to attach to and colonize the cuticular surface of the mouthparts of vectors; a set of recombinant peptides was generated and the chemical affinities of these peptides to chitin and related carbohydrates was assayed in vitro. Two candidates, the X. fastidiosa hypothetical protein PD1764 and an N-terminal region of the hemagglutinin-like protein B (HxfB) showed affinity for these substrates. These proteins were provided to vectors via an artificial diet system in which insects acquire X. fastidiosa, followed by an inoculation access period on plants under greenhouse conditions. Both PD1764 and HxfAD1-3 significantly blocked transmission. Furthermore, bacterial populations within insects over a 10-day period demonstrated that these peptides inhibited cell adhesion to vectors but not bacterial multiplication, indicating that the mode of action of these peptides is restricted to limiting cell adhesion to insects, likely via competition for adhesion sites. These results open a new venue in the search for sustainable disease-control strategies that are pathogen specific and may have limited nontarget effects. PMID:27049684

  18. Test beam performance of CDF plug upgrade EM calorimeter

    SciTech Connect

    Fukui, Y.; CDF Upgrade Group

    1998-01-01

    CDF Plug Upgrade(tile-fiber) EM Calorimeter performed resolution of 15%/{radical}E{circle_plus}0.7% with non-linearity less than 1% in a energy range of 5-180 GeV at Fermilab Test Beam. Transverse uniformity of inside-tower-response of the EM Calorimeter was 2.2% with 56 GeV positron, which was reduced to 1.0% with response map correction. We observed 300 photo electron/GeV in the EM Calorimeter. Ratios of EM Calorimeter response to positron beam to that to {sup 137}Cs Source was stable within 1% in the period of 8 months.

  19. 2. HI PAR (ACQUISITION RADAR) TOWER AND ENLISTED MEN (EM) ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    2. HI PAR (ACQUISITION RADAR) TOWER AND ENLISTED MEN (EM) BARRACKS WITH RADAR ATTACHED. - Nike Hercules Missile Battery Summit Site, Battery Control Administration & Barracks Building, Anchorage, Anchorage, AK

  20. Communication - An Effective Tool for Implementing ISO 14001/EMS

    SciTech Connect

    Rachel Damewood; Bowen Huntsman

    2004-04-01

    The Idaho National Engineering and Environmental Laboratory (INEEL) received ISO 14001/EMS certification in June 2002. Communication played an effective role in implementing ISO 14001/EMS at the INEEL. This paper describes communication strategies used during the implementation and certification processes. The INEEL achieved Integrated Safety Management System (ISMS) and Voluntary Protection Program (VPP) Star status in 2001. ISMS implemented a formal process to plan and execute work. VPP facilitated worker involvement by establishing geographic units at various facilities with employee points of contact and management champions. The INEEL Environmental Management System (EMS) was developed to integrate the environmental functional area into its ISMS and VPP. Since the core functions of ISMS, VPP, and EMS are interchangeable, they were easy to integrate. Communication is essential to successfully implement an EMS. (According to ISO 14001 requirements, communication interacts with 12 other elements of the requirements.) We developed communication strategies that integrated ISMS, VPP, and EMS. For example, the ISMS, VPP, and EMS Web sites communicated messages to the work force, such as “VPP emphasizes the people side of doing business, ISMS emphasizes the system side of doing business, and EMS emphasizes the systems to protect the environment; but they all define work, identify and analyze hazards, and mitigate the hazards.” As a result of this integration, the work force supported and implemented the EMS. In addition, the INEEL established a cross-functional communication team to assist with implementing the EMS. The team included members from the Training and Communication organizations, VPP office, Pollution Prevention, Employee and Media Relations, a union representative, facility environmental support, and EMS staff. This crossfunctional team used various communication strategies to promote our EMS to all organization levels and successfully implemented EMS

  1. EM threat analysis for wireless systems.

    SciTech Connect

    Burkholder, R. J. (Ohio State University Electroscience Laboratory); Mariano, Robert J.; Schniter, P. (Ohio State University Electroscience Laboratory); Gupta, I. J. (Ohio State University Electroscience Laboratory)

    2006-06-01

    Modern digital radio systems are complex and must be carefully designed, especially when expected to operate in harsh propagation environments. The ability to accurately predict the effects of propagation on wireless radio performance could lead to more efficient radio designs as well as the ability to perform vulnerability analyses before and after system deployment. In this report, the authors--experts in electromagnetic (EM) modeling and wireless communication theory--describe the construction of a simulation environment that is capable of quantifying the effects of wireless propagation on the performance of digital communication.

  2. Stochastic EM-based TFBS motif discovery with MITSU

    PubMed Central

    Kilpatrick, Alastair M.; Ward, Bruce; Aitken, Stuart

    2014-01-01

    Motivation: The Expectation–Maximization (EM) algorithm has been successfully applied to the problem of transcription factor binding site (TFBS) motif discovery and underlies the most widely used motif discovery algorithms. In the wider field of probabilistic modelling, the stochastic EM (sEM) algorithm has been used to overcome some of the limitations of the EM algorithm; however, the application of sEM to motif discovery has not been fully explored. Results: We present MITSU (Motif discovery by ITerative Sampling and Updating), a novel algorithm for motif discovery, which combines sEM with an improved approximation to the likelihood function, which is unconstrained with regard to the distribution of motif occurrences within the input dataset. The algorithm is evaluated quantitatively on realistic synthetic data and several collections of characterized prokaryotic TFBS motifs and shown to outperform EM and an alternative sEM-based algorithm, particularly in terms of site-level positive predictive value. Availability and implementation: Java executable available for download at http://www.sourceforge.net/p/mitsu-motif/, supported on Linux/OS X. Contact: a.m.kilpatrick@sms.ed.ac.uk PMID:24931999

  3. CryoEM at IUCrJ: a new era

    PubMed Central

    Subramaniam, Sriram; Kühlbrandt, Werner; Henderson, Richard

    2016-01-01

    In this overview, we briefly outline recent advances in electron cryomicroscopy (cryoEM) and explain why the journal IUCrJ, published by the International Union of Crystallography, could provide a natural home for publications covering many present and future developments in the cryoEM field. PMID:26870375

  4. 7 CFR 1945.20 - Making EM loans available.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... Secretary in making a decision on the requested natural disaster determination. (4) The Secretary will... 7 Agriculture 13 2012-01-01 2012-01-01 false Making EM loans available. 1945.20 Section 1945.20...) PROGRAM REGULATIONS (CONTINUED) EMERGENCY Disaster Assistance-General § 1945.20 Making EM loans...

  5. 7 CFR 1945.20 - Making EM loans available.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... Secretary in making a decision on the requested natural disaster determination. (4) The Secretary will... 7 Agriculture 13 2011-01-01 2009-01-01 true Making EM loans available. 1945.20 Section 1945.20...) PROGRAM REGULATIONS (CONTINUED) EMERGENCY Disaster Assistance-General § 1945.20 Making EM loans...

  6. Click-EM for imaging metabolically tagged nonprotein biomolecules.

    PubMed

    Ngo, John T; Adams, Stephen R; Deerinck, Thomas J; Boassa, Daniela; Rodriguez-Rivera, Frances; Palida, Sakina F; Bertozzi, Carolyn R; Ellisman, Mark H; Tsien, Roger Y

    2016-06-01

    EM has long been the main technique for imaging cell structures with nanometer resolution but has lagged behind light microscopy in the crucial ability to make specific molecules stand out. Here we introduce click-EM, a labeling technique for correlative light microscopy and EM imaging of nonprotein biomolecules. In this approach, metabolic labeling substrates containing bioorthogonal functional groups are provided to cells for incorporation into biopolymers by endogenous biosynthetic machinery. The unique chemical functionality of these analogs is exploited for selective attachment of singlet oxygen-generating fluorescent dyes via bioorthogonal 'click chemistry' ligations. Illumination of dye-labeled structures generates singlet oxygen to locally catalyze the polymerization of diaminobenzidine into an osmiophilic reaction product that is readily imaged by EM. We describe the application of click-EM in imaging metabolically tagged DNA, RNA and lipids in cultured cells and neurons and highlight its use in tracking peptidoglycan synthesis in the Gram-positive bacterium Listeria monocytogenes. PMID:27110681

  7. The Orthogonally Partitioned EM Algorithm: Extending the EM Algorithm for Algorithmic Stability and Bias Correction Due to Imperfect Data.

    PubMed

    Regier, Michael D; Moodie, Erica E M

    2016-05-01

    We propose an extension of the EM algorithm that exploits the common assumption of unique parameterization, corrects for biases due to missing data and measurement error, converges for the specified model when standard implementation of the EM algorithm has a low probability of convergence, and reduces a potentially complex algorithm into a sequence of smaller, simpler, self-contained EM algorithms. We use the theory surrounding the EM algorithm to derive the theoretical results of our proposal, showing that an optimal solution over the parameter space is obtained. A simulation study is used to explore the finite sample properties of the proposed extension when there is missing data and measurement error. We observe that partitioning the EM algorithm into simpler steps may provide better bias reduction in the estimation of model parameters. The ability to breakdown a complicated problem in to a series of simpler, more accessible problems will permit a broader implementation of the EM algorithm, permit the use of software packages that now implement and/or automate the EM algorithm, and make the EM algorithm more accessible to a wider and more general audience. PMID:27227718

  8. Evaluation of Fracture Azimuth by EM Wave and Elastic Wave

    NASA Astrophysics Data System (ADS)

    Feng, X.; Wang, Q.; Liu, C.; Lu, Q.; Zeng, Z.; Liang, W.; Yu, Y.; Ren, Q.

    2013-12-01

    Fracture system plays an important role in the development of underground energy, for example enhanced geothermal system (EGS), oil shale and shale gas, etc. Therefore, it becomes more and more important to detect and evaluate the fracture system. Geophysical prospecting is an useful method to evaluate the characteristics of the subsurface fractures. Currently, micro-seismology, multi-wave seismic exploration, and electromagnetic (EM) survey are reported to be used for the purpose. We are studying a method using both elastic wave and EM wave to detect and evaluate the fracture azimuth in laboratory. First, we build a 3D horizontal transverse isotropy (HTI) model, shown in the figure 1, by dry parallel fractures system, which was constructed by plexiglass plates and papers. Then, we used the ultrasonic system to obtain reflected S-wave data. Depending on the shear wave splitting, we evaluated the fracture azimuth by the algorithm of Pearson correlation coefficient. In addition, we used the full Polarimetric ultra wide band electromagnetic (FP-UWB-EM) wave System, shown in the figure 2, to obtain full polarimetric reflected EM-wave data. Depending on the rotation of the EM wave polarimetry, we evaluated the fracture azimuth by the the ration between maximum amplitude of co-polarimetric EM wave and maximum amplitude of cross-polarimetric EM wave. Finally, we used both EM-wave data and S-wave data to evaluate the fracture azimuth by the method of cross plot and statistical mathematics. To sum up, we found that FP-UWB-EM wave can be used to evaluated the fracture azimuth and is more accurate than ultrasound wave. Also joint evaluation using both data could improve the precision.

  9. Refinement of atomic models in high resolution EM reconstructions using Flex-EM and local assessment

    PubMed Central

    Joseph, Agnel Praveen; Malhotra, Sony; Burnley, Tom; Wood, Chris; Clare, Daniel K.; Winn, Martyn; Topf, Maya

    2016-01-01

    As the resolutions of Three Dimensional Electron Microscopic reconstructions of biological macromolecules are being improved, there is a need for better fitting and refinement methods at high resolutions and robust approaches for model assessment. Flex-EM/MODELLER has been used for flexible fitting of atomic models in intermediate-to-low resolution density maps of different biological systems. Here, we demonstrate the suitability of the method to successfully refine structures at higher resolutions (2.5–4.5 Å) using both simulated and experimental data, including a newly processed map of Apo-GroEL. A hierarchical refinement protocol was adopted where the rigid body definitions are relaxed and atom displacement steps are reduced progressively at successive stages of refinement. For the assessment of local fit, we used the SMOC (segment-based Manders’ overlap coefficient) score, while the model quality was checked using the Qmean score. Comparison of SMOC profiles at different stages of refinement helped in detecting regions that are poorly fitted. We also show how initial model errors can have significant impact on the goodness-of-fit. Finally, we discuss the implementation of Flex-EM in the CCP-EM software suite. PMID:26988127

  10. Refinement of atomic models in high resolution EM reconstructions using Flex-EM and local assessment.

    PubMed

    Joseph, Agnel Praveen; Malhotra, Sony; Burnley, Tom; Wood, Chris; Clare, Daniel K; Winn, Martyn; Topf, Maya

    2016-05-01

    As the resolutions of Three Dimensional Electron Microscopic reconstructions of biological macromolecules are being improved, there is a need for better fitting and refinement methods at high resolutions and robust approaches for model assessment. Flex-EM/MODELLER has been used for flexible fitting of atomic models in intermediate-to-low resolution density maps of different biological systems. Here, we demonstrate the suitability of the method to successfully refine structures at higher resolutions (2.5-4.5Å) using both simulated and experimental data, including a newly processed map of Apo-GroEL. A hierarchical refinement protocol was adopted where the rigid body definitions are relaxed and atom displacement steps are reduced progressively at successive stages of refinement. For the assessment of local fit, we used the SMOC (segment-based Manders' overlap coefficient) score, while the model quality was checked using the Qmean score. Comparison of SMOC profiles at different stages of refinement helped in detecting regions that are poorly fitted. We also show how initial model errors can have significant impact on the goodness-of-fit. Finally, we discuss the implementation of Flex-EM in the CCP-EM software suite. PMID:26988127

  11. Environmental Education and Development Division (EM-522). Annual report, Fiscal year 1993

    SciTech Connect

    Not Available

    1993-12-31

    The Environmental Education and Development Division (EM-522) is one of three divisions within the Office of Technology Integration and Environmental Education and Development (EM-52) in Environmental Restoration and Waste Management`s (EM`s) Office of Technology Development (EM-50). The primary design criterion for EM-522 education activities is directly related to meeting EM`s goal of environmental compliance on an accelerated basis and cleanup of the 1989 inventory of inactive sites and facilities by the year 2019. Therefore, EM-522`s efforts are directed specifically toward stimulating knowledge and capabilities to achieve the goals of EM while contributing to DOE`s overall goal of increasing scientific, mathematical, and technical literacy and competency. This report discusses fiscal year 1993 activities.

  12. Electromagnetic optimization of EMS-MAGLEV systems

    SciTech Connect

    Andriollo, M.; Martinelli, G.; Morini, A.; Tortella, A.

    1998-07-01

    In EMS-MAGLEV high-speed transport systems, devices for propulsion, levitation and contactless on-board electric power transfer are combined in a single electromagnetic structure. The strong coupling among the windings affects the performance of each device and requires the utilization of numerical codes. The paper describes an overall optimization procedure, based on a suitable mathematical model of the system, which takes into account several items of the system performance. The parameters of the model are calculated by an automated sequence of FEM analyses of the configuration. Both the linear generator output characteristics and the propulsion force ripple are improved applying the procedure to a reference configuration. The results are compared with the results obtained by a sequence of partial optimizations operating separately on two different subsets of the geometric parameters.

  13. DOE EM industry programs robotics development

    SciTech Connect

    Staubly, R.; Kothari, V.

    1997-12-01

    The Office of Science and Technology (OST) manages an aggressive program for RD&D, as well as testing and evaluation for the U.S. Department of Energy`s (DOE) Environmental Management (EM) organization. The goal is to develop new and improved environmental restoration and waste management technologies to clean up the inventory of the DOE weapons complex faster, safer, and cheaper than is possible with currently available technologies. OST has organized technology management activities along focus teams for each major problem area. There are currently five focus areas: decontamination and decommissioning, tanks, subsurface contaminants, mixed waste, and plutonium. In addition, OST is pursuing research and development (R&D) that cuts across these focus areas by having applications in two or more focus areas. Currently, there are three cross-cutting programs: the robotics technology development; characterization, monitoring, and sensor technologies; and efficient separations and processing.

  14. Online EM with weight-based forgetting.

    PubMed

    Celaya, Enric; Agostini, Alejandro

    2015-05-01

    In the online version of the EM algorithm introduced by Sato and Ishii ( 2000 ), a time-dependent discount factor is introduced for forgetting the effect of the old estimated values obtained with an earlier, inaccurate estimator. In their approach, forgetting is uniformly applied to the estimators of each mixture component depending exclusively on time, irrespective of the weight attributed to each unit for the observed sample. This causes an excessive forgetting in the less frequently sampled regions. To address this problem, we propose a modification of the algorithm that involves a weight-dependent forgetting, different for each mixture component, in which old observations are forgotten according to the actual weight of the new samples used to replace older values. A comparison of the time-dependent versus the weight-dependent approach shows that the latter improves the accuracy of the approximation and exhibits much greater stability. PMID:25710091

  15. Processing of Structurally Heterogeneous Cryo-EM Data in RELION.

    PubMed

    Scheres, S H W

    2016-01-01

    This chapter describes algorithmic advances in the RELION software, and how these are used in high-resolution cryo-electron microscopy (cryo-EM) structure determination. Since the presence of projections of different three-dimensional structures in the dataset probably represents the biggest challenge in cryo-EM data processing, special emphasis is placed on how to deal with structurally heterogeneous datasets. As such, this chapter aims to be of practical help to those who wish to use RELION in their cryo-EM structure determination efforts. PMID:27572726

  16. Validation of real-time PCR assays for bioforensic detection of model plant pathogens.

    PubMed

    James, Mindy; Blagden, Trenna; Moncrief, Ian; Burans, James P; Schneider, Katherine; Fletcher, Jacqueline

    2014-03-01

    The U.S. agricultural sector is vulnerable to intentionally introduced microbial threats because of its wide and open distribution and economic importance. To investigate such events, forensically valid assays for plant pathogen detection are needed. In this work, real-time PCR assays were developed for three model plant pathogens: Pseudomonas syringae pathovar tomato, Xylella fastidiosa, and Wheat streak mosaic virus. Validation included determination of the linearity and range, limit of detection, sensitivity, specificity, and exclusivity of each assay. Additionally, positive control plasmids, distinguishable from native signature by restriction enzyme digestion, were developed to support forensic application of the assays. Each assay displayed linear amplification of target nucleic acid, detected 100 fg or less of target nucleic acid, and was specific to its target pathogen. Results obtained with these model pathogens provide the framework for development and validation of similar assays for other plant pathogens of high consequence. PMID:24261870

  17. RNAi effects on actin mRNAs in Homalodisca vitripennis cells

    PubMed Central

    Rosa, Cristina; Kamita, Shizuo G; Dequine, Haley; Wuriyanghan, Ha; Lindbo, John A; Falk, Bryce W

    2010-01-01

    The xylem feeding leafhopper Homalodisaca vitripennis (H. vitripennis) is an unusually robust and efficient vector of Xylella fastidiosa, a Gram-negative bacterium which causes several very important plant diseases. Here we investigated RNA interference (RNAi) to target actin, a key component of insect cells and whole bodies, in H. vitripennis cells. RNAi effectors were delivered via lipid based transfection and real-time RT-PCR, RNA hybridization, and microscopic analyses were employed to verify RNAi effects. When actin dsRNAs were used, a 10-fold decrease in the target H. vitripennis actin mRNA level was seen in cells. Altered phenotypic effects also were evident in transfected cells, as were small interfering RNAs, hallmarks of RNAi. The use of H. vitripennis cells and RNAi offers new opportunities to research hemipterans, the most important insect vectors of plant pathogens. PMID:20628496

  18. Pattern formation exhibited by biofilm formation within microfluidic chambers.

    PubMed

    Cogan, N G; Donahue, M R; Whidden, Mark; De La Fuente, Leonardo

    2013-05-01

    This article investigates the dynamics of an important bacterial pathogen, Xylella fastidiosa, within artificial plant xylem. The bacterium is the causative agent of a variety of diseases that strike fruit-bearing plants including Pierce's disease of grapevine. Biofilm colonization within microfluidic chambers was visualized in a laboratory setting, showing robust, regular spatial patterning. We also develop a mathematical model, based on a multiphase approach that is able to capture the spacing of the pattern and points to the role of the exopolymeric substance as the main source of control of the pattern dynamics. We concentrate on estimating the attachment/detachment processes within the chamber because these are two mechanisms that have the potential to be engineered by applying various chemicals to prevent or treat the disease. PMID:23663829

  19. Pattern Formation Exhibited by Biofilm Formation within Microfluidic Chambers

    PubMed Central

    Cogan, N.G.; Donahue, M.R.; Whidden, Mark; De La Fuente, Leonardo

    2013-01-01

    This article investigates the dynamics of an important bacterial pathogen, Xylella fastidiosa, within artificial plant xylem. The bacterium is the causative agent of a variety of diseases that strike fruit-bearing plants including Pierce’s disease of grapevine. Biofilm colonization within microfluidic chambers was visualized in a laboratory setting, showing robust, regular spatial patterning. We also develop a mathematical model, based on a multiphase approach that is able to capture the spacing of the pattern and points to the role of the exopolymeric substance as the main source of control of the pattern dynamics. We concentrate on estimating the attachment/detachment processes within the chamber because these are two mechanisms that have the potential to be engineered by applying various chemicals to prevent or treat the disease. PMID:23663829

  20. Test beam performance of CDF plug upgrade EM calorimeter

    SciTech Connect

    Fukui, Y.

    1998-11-01

    CDF Plug Upgrade(tile-fiber) EM Calorimeter performed resolution of 15{percent}/{radical} (E) {circle_plus}0.7{percent} with non-linearity less than 1{percent} in a energy range of 5{endash}180 GeV at Fermilab Test Beam. Transverse uniformity of inside-tower-response of the EM Calorimeter was 2.2{percent} with 56 GeV positron, which was reduced to 1.0{percent} with response map correction. We observed 300 photo electron/GeV in the EM Calorimeter. Ratios of EM Calorimeter response to positron beam to that to {sup 137}C{sub s} Source was stable within 1{percent} in the period of 8 months. {copyright} {ital 1998 American Institute of Physics.}

  1. E.M. and Hadronic Shower Simulation with FLUKA

    SciTech Connect

    Battistoni, G.; Fasso, A.; Ferrari, A.; Ranft, J.; Rubbia, A.; Sala, P.R.; /INFN, Milan /SLAC /CERN /Siegen U. /Zurich, ETH

    2005-10-03

    A description of the main features of e.m. and hadronic shower simulation models used in the FLUKA code is summarized and some recent applications are discussed. The general status of the FLUKA project is also reported.

  2. 7 CFR 759.6 - EM to be made available.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... Agriculture under the Plant Protection Act or the animal quarantine laws, as defined in section 2509 of the Food, Agriculture, Conservation, and Trade Act of 1990, automatically authorizes EM for production...

  3. 7 CFR 759.6 - EM to be made available.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... Agriculture under the Plant Protection Act or the animal quarantine laws, as defined in section 2509 of the Food, Agriculture, Conservation, and Trade Act of 1990, automatically authorizes EM for production...

  4. NASA EM Followup of LIGO-Virgo Candidate Events

    NASA Technical Reports Server (NTRS)

    Blackburn, Lindy L.

    2011-01-01

    We present a strategy for a follow-up of LIGO-Virgo candidate events using offline survey data from several NASA high-energy photon instruments aboard RXTE, Swift, and Fermi. Time and sky-location information provided by the GW trigger allows for a targeted search for prompt and afterglow EM signals. In doing so, we expect to be sensitive to signals which are too weak to be publicly reported as astrophysical EM events.

  5. EM Telemetry Tool for Deep Well Drilling Applications

    SciTech Connect

    Jeffrey M. Gabelmann

    2005-11-15

    This final report discusses the successful development and testing of a deep operational electromagnetic (EM) telemetry system, produced under a cooperative agreement with the United States Department of Energy's National Energy Technology Laboratory. This new electromagnetic telemetry system provides a wireless communication link between sensors deployed deep within oil and gas wells and data acquisition equipment located on the earth's surface. EM based wireless telemetry is a highly appropriate technology for oil and gas exploration in that it avoids the need for thousands of feet of wired connections. In order to achieve the project performance objectives, significant improvements over existing EM telemetry systems were made. These improvements included the development of new technologies that have improved the reliability of the communications link while extending operational depth. A key element of the new design is the incorporation of a data-fusion methodology which enhances the communication receiver's ability to extract very weak signals from large amounts of ambient environmental noise. This innovative data-fusion receiver based system adapts advanced technologies, not normally associated with low-frequency communications, and makes them work within the harsh drilling environments associated with the energy exploration market. Every element of a traditional EM telemetry system design, from power efficiency to reliability, has been addressed. The data fusion based EM telemetry system developed during this project is anticipated to provide an EM tool capability that will impact both onshore and offshore oil and gas exploration operations, for conventional and underbalanced drilling applications.

  6. A HF EM installation allowing simultaneous whole body and deep local EM hyperthermia.

    PubMed

    Mazokhin, V N; Kolmakov, D N; Lucheyov, N A; Gelvich, E A; Troshin, I I

    1999-01-01

    The structure and main features of a HF EM installation based upon a new approach for creating electromagnetic fields destined for whole body (WBH) and deep local (DLH) hyperthermia are discussed. The HF EM field, at a frequency of 13.56 MHz, is created by a coplanar capacity type applicator positioned under a distilled water filled bolus that the patient is lying on. The EM energy being released directly in the deep tissues ensures effective whole body heating to required therapeutic temperatures of up to 43.5 degrees C, whereas the skin temperature can be maintained as low as 39-40.5 degrees C. For DLH, the installation is equipped with additional applicators and a generator operating at a frequency of 40.68 MHz. High efficiency of the WBH applicator makes it possible to carry out the WBH procedure without any air-conditioning cabin. Due to this, a free access to the patient's body during the WBH treatment is provided and a simultaneous WBH/DLH or WBH/LH procedure by means of additional applicators is possible. Controllable power output in the range of 100-800 W at a frequency of 13.56 MHz and 50-350 W at a frequency of 40.68 MHz allows accurate temperature control during WBH, DLH and WBH/DLH procedures. SAR patterns created by the WBH and DLH applicators in a liquid muscle phantom and measured by means of a non-perturbing E-dipole are investigated. The scattered EM field strength measured in the vicinity of the operating installation during the WBH, DLH and WBH/DLH procedures does not exceed security standards. Examples of temperature versus time graphs in the course of WBH, DLH and WBH/DLH procedures in clinics are presented. The installation is successfully used in leading oncological institutions of Russia and Belarus, though combined WBH/DLH procedures are evidently more complicated and demand thorough planning and temperature measurements to avoid overheating. PMID:10458570

  7. Analysis of EM dataset with several sensor configurations obtained by the loop-loop EM survey on magnetic anomalies

    NASA Astrophysics Data System (ADS)

    CHOI, J.; Yi, M. J.; Sasaki, Y.; Son, J.; Nam, M. J.

    2015-12-01

    Most of mineral mines in Korea are located in rugged mountain area embedding small-scale anomalies. Loop-loop EM survey system can be a better choice for exploring those mines because no ground contact is required and portable loops are freely positioned. Survey design is very important for detecting small amount of mineral deposits efficiently and spatial limits of survey lines should be considered. Along a same survey line, surveys with different separations between a transmitter and a receiver are applicable. EM responses are calculated in a layered-earth model embedding magnetic anomalies and analyses considering electric conductivity and magnetic permeability are made for the loop-loop EM survey data. Combining EM dataset with multi-frequency and multi-separation slightly enhanced a reconstructed image. Loop-loop EM survey using PROMOIS system was conducted on a small magnetite mine. Inversion with and without considering magnetic permeability was conducted for EM data with multi-frequency and multi-separation between a transmitter and a receiver.

  8. DOE EM industry programs robotics development

    SciTech Connect

    Staubly, R.; Kothari, V.

    1998-12-31

    The Office of Science and Technology (OST) manages an aggressive program for RD and D, as well as testing and evaluation for the Department of Energy`s (DOE`s) Environmental Management (EM) organization. The goal is to develop new and improved environmental restoration and waste management technologies to clean up the inventory of the DOE weapons complex faster, safer, and cheaper than is possible with currently available technologies. Robotic systems reduce worker exposure to the absolute minimum, while providing proven, cost-effective, and, for some applications, the only acceptable technique for addressing challenging problems. Development of robotic systems for remote operations occurs in three main categories: tank waste characterization and retrieval; decontamination and dismantlement; and characterization, mapping, and inspection systems. In addition, the Federal Energy Technology Center (FETC) has some other projects which fall under the heading of supporting R and D. The central objective of all FETC robotic projects is to make robotic systems more attractive by reducing costs and health risks associated with the deployment of robotic technologies in the cleanup of the nuclear weapons complex. This will be accomplished through development of robots that are cheaper, faster, safer, and more reliable, as well as more straightforward to modify/adapt and more intuitive to operate with autonomous capabilities and intelligent controls that prevent accidents and optimize task execution.

  9. Databases and Archiving for CryoEM.

    PubMed

    Patwardhan, A; Lawson, C L

    2016-01-01

    CryoEM in structural biology is currently served by three public archives-EMDB for 3DEM reconstructions, PDB for models built from 3DEM reconstructions, and EMPIAR for the raw 2D image data used to obtain the 3DEM reconstructions. These archives play a vital role for both the structural community and the wider biological community in making the data accessible so that results may be reused, reassessed, and integrated with other structural and bioinformatics resources. The important role of the archives is underpinned by the fact that many journals mandate the deposition of data to PDB and EMDB on publication. The field is currently undergoing transformative changes where on the one hand high-resolution structures are becoming a routine occurrence while on the other hand electron tomography is enabling the study of macromolecules in the cellular context. Concomitantly the archives are evolving to best serve their stakeholder communities. In this chapter, we describe the current state of the archives, resources available for depositing, accessing, searching, visualizing and validating data, on-going community-wide initiatives and opportunities, and challenges for the future. PMID:27572735

  10. Degradation of Benzodiazepines after 120 Days of EMS Deployment

    PubMed Central

    McMullan, Jason T.; Jones, Elizabeth; Barnhart, Bruce; Denninghoff, Kurt; Spaite, Daniel; Zaleski, Erin; Silbergleit, Robert

    2014-01-01

    Introduction EMS treatment of status epilepticus improves outcomes, but the benzodiazepine best suited for EMS use is unclear, given potential high environmental temperature exposures. Objective To describe the degradation of diazepam, lorazepam, and midazolam as a function of temperature exposure and time over 120 days of storage on active EMS units. Methods Study boxes containing vials of diazepam, lorazepam, and midazolam were distributed to 4 active EMS units in each of 2 EMS systems in the southwestern United States during May–August 2011. The boxes logged temperature every minute and were stored in EMS units per local agency policy. Two vials of each drug were removed from each box at 30-day intervals and underwent high-performance liquid chromatography to determine drug concentration. Concentration was analyzed as mean (and 95%CI) percent of initial labeled concentration as a function of time and mean kinetic temperature (MKT). Results 192 samples were collected (2 samples of each drug from each of 4 units per city at 4 time-points). After 120 days, the mean relative concentration (95%CI) of diazepam was 97.0% (95.7–98.2%) and of midazolam was 99.0% (97.7–100.2%). Lorazepam experienced modest degradation by 60 days (95.6% [91.6–99.5%]) and substantial degradation at 90 days (90.3% [85.2-95.4%]) and 120 days (86.5% [80.7–92.3%]). Mean MKT was 31.6°C (95%CI 27.1–36.1). Increasing MKT was associated with greater degradation of lorazepam, but not midazolam or diazepam. Conclusions Midazolam and diazepam experienced minimal degradation throughout 120 days of EMS deployment in high-heat environments. Lorazepam experienced significant degradation over 120 days and appeared especially sensitive to higher MKT exposure. PMID:24548058

  11. Emergency medical service (EMS): A unique flight environment

    NASA Technical Reports Server (NTRS)

    Shively, R. Jay

    1993-01-01

    The EMS flight environment is unique in today's aviation. The pilots must respond quickly to emergency events and often fly to landing zones where they have never been before . The time from initially receiving a call to being airborne can be as little as two to three minutes. Often the EMS pilot is the only aviation professional on site, they have no operations people or other pilots to aid them in making decisons. Further, since they are often flying to accident scenes, not airports, there is often complete weather and condition information. Therefore, the initial decision that the pilot must make, accepting or declining a flight, can become very difficult. The accident rate of EMS helicopters has been relatively high over the past years. NASA-Ames research center has taken several steps in an attempt to aid EMS pilots in their decision making and situational awareness. A preflight risk assessment system (SAFE) was developed to aid pilots in their decision making, and was tested at an EMS service. The resutls of the study were promising and a second version incorporating the lessons learned is under development. A second line of research was the development of a low cost electronic chart display (ECD). This is a digital map display to help pilots maintain geographical orientation. Another thrust was undertaken in conjunction with the Aviation Safety Reporting System (ASRS). This involved publicizing the ASRS to EMS pilots and personnel, and calling each of the reporters back to gather additional information. This paper will discuss these efforts and how they may positively impact the safety of EMS operations.

  12. Persistent topology for cryo-EM data analysis.

    PubMed

    Xia, Kelin; Wei, Guo-Wei

    2015-08-01

    In this work, we introduce persistent homology for the analysis of cryo-electron microscopy (cryo-EM) density maps. We identify the topological fingerprint or topological signature of noise, which is widespread in cryo-EM data. For low signal-to-noise ratio (SNR) volumetric data, intrinsic topological features of biomolecular structures are indistinguishable from noise. To remove noise, we employ geometric flows that are found to preserve the intrinsic topological fingerprints of cryo-EM structures and diminish the topological signature of noise. In particular, persistent homology enables us to visualize the gradual separation of the topological fingerprints of cryo-EM structures from those of noise during the denoising process, which gives rise to a practical procedure for prescribing a noise threshold to extract cryo-EM structure information from noise contaminated data after certain iterations of the geometric flow equation. To further demonstrate the utility of persistent homology for cryo-EM data analysis, we consider a microtubule intermediate structure Electron Microscopy Data (EMD 1129). Three helix models, an alpha-tubulin monomer model, an alpha-tubulin and beta-tubulin model, and an alpha-tubulin and beta-tubulin dimer model, are constructed to fit the cryo-EM data. The least square fitting leads to similarly high correlation coefficients, which indicates that structure determination via optimization is an ill-posed inverse problem. However, these models have dramatically different topological fingerprints. Especially, linkages or connectivities that discriminate one model from another, play little role in the traditional density fitting or optimization but are very sensitive and crucial to topological fingerprints. The intrinsic topological features of the microtubule data are identified after topological denoising. By a comparison of the topological fingerprints of the original data and those of three models, we found that the third model is

  13. Colloidal Oatmeal <em>(Avena Sativa)em> Improves Skin Barrier Through Multi-Therapy Activity.

    PubMed

    Ilnytska, Olha; Kaur, Simarna; Chon, Suhyoun; Reynertson, Kurt A; Nebus, Judith; Garay, Michelle; Mahmood, Khalid; Southall, Michael D

    2016-06-01

    Oats (Avena sativa) are a centuries-old topical treatment for a variety of skin barrier conditions, including dry skin, skin rashes, and eczema; however, few studies have investigated the actual mechanism of action for the skin barrier strengthening activity of colloidal oatmeal. Four extracts of colloidal oatmeal were prepared with various solvents and tested in vitro for skin barrier related gene expression and activity. Extracts of colloidal oatmeal were found to induce the expression of genes related to epidermal differentiation, tight junctions and lipid regulation in skin, and provide pH-buffering capacity. Colloidal oatmeal boosted the expression of multiple target genes related to skin barrier, and resulted in recovery of barrier damage in an in vitro model of atopic dermatitis. In addition, an investigator-blinded study was performed with 50 healthy female subjects who exhibited bilateral moderate to severe dry skin on their lower legs. Subjects were treated with a colloidal oatmeal skin protectant lotion. Clinically, the colloidal oatmeal lotion showed significant clinical improvements in skin dryness, moisturization, and barrier. Taken together, these results demonstrate that colloidal oatmeal can provide clinically effective benefits for dry and compromised skin by strengthening skin barrier.

    <em>J Drugs Dermatolem>. 2016;15(6):684-690. PMID:27272074

  14. Learning when to Hold'em and When to Fold'em: ERS's Budget Hold'em Game Facilitates the Budget Development Process in Memphis

    ERIC Educational Resources Information Center

    Education Resource Strategies, 2010

    2010-01-01

    If your school district is facing a budget issue, it might surprise you to learn that the solution might very well lie in a game of cards. That certainly was the case earlier this year for the city schools of Memphis, Tennessee. The game is called Budget Hold'em, and it was developed by Education Resource Strategies (ERS) of Watertown,…

  15. Topical Treatment With an Agent Disruptive to <em>P. acnesem> Biofilm Provides Positive Therapeutic Response: Results of a Randomized Clinical Trial.

    PubMed

    Bernhardt, Michael J; Myntti, Matthew F

    2016-06-01

    The traditional disease model of acne has been one of follicular plugging due to 'sticky epithelial cells' associated with increased sebum production with deep follicular anaerobic conditions favoring <em>P. acnesem>- generated inflammation. <em>P. acnesem> biofilms have been found more frequently in patients with acne than controls. Biofilms are genetically coded to create adhesion to the pilosebaceous unit followed by production of a mucopolysaccharide coating capable of binding to lipid surfaces. Traditional therapies for acne have involved mixtures of oral and topical antibiotics admixed with topical keratolytics and retinoids, which are aimed at traditional bacterial reduction as well as downregulating the inflammatory cascade. These approaches are limited by side effect and compliance/tolerability issues. As the <em>P. acnesem> biofilm may, in fact, be the instigator of this process, we studied the use of a topical agent designed to reduce the <em>P. acnesem> biofilm to see if reducing the biofilm would be therapeutically efficacious. We present data of a proprietary topical non-prescription agent with a novel pharmaco mechanism designed to attack the biofilm produced by <em>P. acnesem>. Our data shows a decrease of inflammatory lesions by 44% and non-inflammatory lesions by 32% after 12 weeks and also provided for a meaningful improvement in the quality of life of the patients in the study. These improvements were achieved with a product that was not associated with burning, chafing, irritation, or erythema, which can be seen with topical treatments. It is apparent from this study that by addressing the biofilm which protects the <em>P. acnesem> bacteria through the use of the Acne Gel, the incidence of acne symptoms can be greatly reduced, while having no negative impacts on the patients' skin (ClinicalTrials.gov registry number NCT02404285).

    <em>J Drugs Dermatol. em>2016;15(6):677-683. PMID:27272073

  16. Refinement of Atomic Structures Against cryo-EM Maps.

    PubMed

    Murshudov, G N

    2016-01-01

    This review describes some of the methods for atomic structure refinement (fitting) against medium/high-resolution single-particle cryo-EM reconstructed maps. Some of the tools developed for macromolecular X-ray crystal structure analysis, especially those encapsulating prior chemical and structural information can be transferred directly for fitting into cryo-EM maps. However, despite the similarities, there are significant differences between data produced by these two techniques; therefore, different likelihood functions linking the data and model must be used in cryo-EM and crystallographic refinement. Although tools described in this review are mostly designed for medium/high-resolution maps, if maps have sufficiently good quality, then these tools can also be used at moderately low resolution, as shown in one example. In addition, the use of several popular crystallographic methods is strongly discouraged in cryo-EM refinement, such as 2Fo-Fc maps, solvent flattening, and feature-enhanced maps (FEMs) for visualization and model (re)building. Two problems in the cryo-EM field are overclaiming resolution and severe map oversharpening. Both of these should be avoided; if data of higher resolution than the signal are used, then overfitting of model parameters into the noise is unavoidable, and if maps are oversharpened, then at least parts of the maps might become very noisy and ultimately uninterpretable. Both of these may result in suboptimal and even misleading atomic models. PMID:27572731

  17. Active geophysical monitoring of hydrocarbon reservoirs using EM methods

    NASA Astrophysics Data System (ADS)

    Gribenko, A.; Black, N.; Zhdanov, M. S.

    2008-12-01

    Marine controlled-source electromagnetic (MCSEM) technology has been successfully established as an effective tool for offshore hydrocarbon (HC) exploration. In this paper we consider another application of the MCSEM method for HC reservoir monitoring. We demonstrate that EM methods can be successfully used for the monitoring of producing wells in connection with the enhanced recovery of hydrocarbons. We have developed a new powerful EM modeling technique based on the integral equation method with an inhomogeneous background conductivity (IE IBC). This new method and the corresponding computer software make it possible to model the EM response over a realistic complex model of a sea-bottom HC reservoir. The numerical modeling results demonstrate that the MCSEM method has the ability to map changes in resistivity caused by the production of hydrocarbons over time. In addition, the EM data help to visualize the changes in the location of the oil-water contact within the reservoir. This result opens the possibility for practical application of the EM method in HC reservoir monitoring.

  18. Application of the EM algorithm to radiographic images.

    PubMed

    Brailean, J C; Little, D; Giger, M L; Chen, C T; Sullivan, B J

    1992-01-01

    The expectation maximization (EM) algorithm has received considerable attention in the area of positron emitted tomography (PET) as a restoration and reconstruction technique. In this paper, the restoration capabilities of the EM algorithm when applied to radiographic images is investigated. This application does not involve reconstruction. The performance of the EM algorithm is quantitatively evaluated using a "perceived" signal-to-noise ratio (SNR) as the image quality metric. This perceived SNR is based on statistical decision theory and includes both the observer's visual response function and a noise component internal to the eye-brain system. For a variety of processing parameters, the relative SNR (ratio of the processed SNR to the original SNR) is calculated and used as a metric to compare quantitatively the effects of the EM algorithm with two other image enhancement techniques: global contrast enhancement (windowing) and unsharp mask filtering. The results suggest that the EM algorithm's performance is superior when compared to unsharp mask filtering and global contrast enhancement for radiographic images which contain objects smaller than 4 mm. PMID:1435595

  19. Breaking Cryo-EM Resolution Barriers to Facilitate Drug Discovery.

    PubMed

    Merk, Alan; Bartesaghi, Alberto; Banerjee, Soojay; Falconieri, Veronica; Rao, Prashant; Davis, Mindy I; Pragani, Rajan; Boxer, Matthew B; Earl, Lesley A; Milne, Jacqueline L S; Subramaniam, Sriram

    2016-06-16

    Recent advances in single-particle cryoelecton microscopy (cryo-EM) are enabling generation of numerous near-atomic resolution structures for well-ordered protein complexes with sizes ≥ ∼200 kDa. Whether cryo-EM methods are equally useful for high-resolution structural analysis of smaller, dynamic protein complexes such as those involved in cellular metabolism remains an important question. Here, we present 3.8 Å resolution cryo-EM structures of the cancer target isocitrate dehydrogenase (93 kDa) and identify the nature of conformational changes induced by binding of the allosteric small-molecule inhibitor ML309. We also report 2.8-Å- and 1.8-Å-resolution structures of lactate dehydrogenase (145 kDa) and glutamate dehydrogenase (334 kDa), respectively. With these results, two perceived barriers in single-particle cryo-EM are overcome: (1) crossing 2 Å resolution and (2) obtaining structures of proteins with sizes < 100 kDa, demonstrating that cryo-EM can be used to investigate a broad spectrum of drug-target interactions and dynamic conformational states. PMID:27238019

  20. International Space Station (ISS) Emergency Mask (EM) Development

    NASA Technical Reports Server (NTRS)

    Toon, Katherine P.; Hahn, Jeffrey; Fowler, Michael; Young, Kevin

    2011-01-01

    The Emergency Mask (EM) is considered a secondary response emergency Personal Protective Equipment (PPE) designed to provide respiratory protection to the International Space Station (ISS) crewmembers in response to a post-fire event or ammonia leak. The EM is planned to be delivered to ISS in 2012 to replace the current air purifying respirator (APR) onboard ISS called the Ammonia Respirator (AR). The EM is a one ]size ]fits ]all model designed to fit any size crewmember, unlike the APR on ISS, and uses either two Fire Cartridges (FCs) or two Commercial Off-the-Shelf (COTS) 3M(Trademark). Ammonia Cartridges (ACs) to provide the crew with a minimum of 8 hours of respiratory protection with appropriate cartridge swap ]out. The EM is designed for a single exposure event, for either post ]fire or ammonia, and is a passive device that cannot help crewmembers who cannot breathe on their own. The EM fs primary and only seal is around the wearer fs neck to prevent a crewmember from inhaling contaminants. During the development of the ISS Emergency Mask, several design challenges were faced that focused around manufacturing a leak free mask. The description of those challenges are broadly discussed but focuses on one key design challenge area: bonding EPDM gasket material to Gore(Registered Trademark) fabric hood.

  1. [Faunistic analysis of leafhopper (Hemiptera: Cicadellidae) species in vineyards of Rio Grande do Sul State, Brazil].

    PubMed

    Ringenberg, Rudiney; Lopes, João R S; Botton, Marcos; Azevedo-Filho, Wilson S De; Cavichioli, Rodney R

    2010-01-01

    In some American countries, grapevines are affected by Pierce's disease (PD), which is caused by a particular strain of Xylella fastidiosa not yet reported in Brazil. In order to investigate the potential for PD spread in Brazil in case of pathogen introduction, we conducted a faunistic analysis of leafhoppers (Hemiptera: Cicadellidae) associated to vineyards in the State of Rio Grande do Sul, with emphasis in the subfamily Cicadellinae (sharpshooters), which includes the main potential vectors of X. fastidiosa. Leafhopper samplings were carried out fortnightly during two years (9/2004-9/2006) in four Vitis vinifera vineyards in the municipalities of Bento Gonçalves and Farroupilha (RS). Thirtyfour leafhopper and six spittlebug species were collected, but most (98.4%) of the 3,893 specimens trapped were leafhoppers, distributed in the subfamilies Cicadellinae (60.2%), Gyponinae (34.1%), Deltocephalinae (3.8%) and Coelidinae (0.3%). The sharpshooter specimens were divided in the tribes Cicadellini (68.5%; 12 species) and Proconiini (31.5%; 11 species). Based on the faunistic indices, five species of Cicadellini, Bucephalogonia xanthophis (Berg), Dilobopterus dispar (Germar), Macugonalia cavifrons Stal, Sibovia sagata (Signoret) and Spinagonalia rubrovittata Cavichioli, and three of Proconiini, Molomea consolida (Schöder), Oncometopia facialis (Signoret) and Oncometopia fusca Melichar were prevalent in the vineyards. The high diversity of native sharpshooters in Rio Grande do Sul indicates the existence of a high risk of PD spread if the pathogen is introduced in grapevines. PMID:20498954

  2. An engineered innate immune defense protects grapevines from Pierce disease.

    PubMed

    Dandekar, Abhaya M; Gouran, Hossein; Ibáñez, Ana María; Uratsu, Sandra L; Agüero, Cecilia B; McFarland, Sarah; Borhani, Yasmin; Feldstein, Paul A; Bruening, George; Nascimento, Rafael; Goulart, Luiz R; Pardington, Paige E; Chaudhary, Anu; Norvell, Meghan; Civerolo, Edwin; Gupta, Goutam

    2012-03-01

    We postulated that a synergistic combination of two innate immune functions, pathogen surface recognition and lysis, in a protein chimera would lead to a robust class of engineered antimicrobial therapeutics for protection against pathogens. In support of our hypothesis, we have engineered such a chimera to protect against the gram-negative Xylella fastidiosa (Xf), which causes diseases in multiple plants of economic importance. Here we report the design and delivery of this chimera to target the Xf subspecies fastidiosa (Xff), which causes Pierce disease in grapevines and poses a great threat to the wine-growing regions of California. One domain of this chimera is an elastase that recognizes and cleaves MopB, a conserved outer membrane protein of Xff. The second domain is a lytic peptide, cecropin B, which targets conserved lipid moieties and creates pores in the Xff outer membrane. A flexible linker joins the recognition and lysis domains, thereby ensuring correct folding of the individual domains and synergistic combination of their functions. The chimera transgene is fused with an amino-terminal signal sequence to facilitate delivery of the chimera to the plant xylem, the site of Xff colonization. We demonstrate that the protein chimera expressed in the xylem is able to directly target Xff, suppress its growth, and significantly decrease the leaf scorching and xylem clogging commonly associated with Pierce disease in grapevines. We believe that similar strategies involving protein chimeras can be developed to protect against many diseases caused by human and plant pathogens. PMID:22355130

  3. An engineered innate immune defense protects grapevines from Pierce disease

    PubMed Central

    Dandekar, Abhaya M.; Gouran, Hossein; Ibáñez, Ana María; Uratsu, Sandra L.; Agüero, Cecilia B.; McFarland, Sarah; Borhani, Yasmin; Feldstein, Paul A.; Bruening, George; Nascimento, Rafael; Goulart, Luiz R.; Pardington, Paige E.; Chaudhary, Anu; Norvell, Meghan; Civerolo, Edwin; Gupta, Goutam

    2012-01-01

    We postulated that a synergistic combination of two innate immune functions, pathogen surface recognition and lysis, in a protein chimera would lead to a robust class of engineered antimicrobial therapeutics for protection against pathogens. In support of our hypothesis, we have engineered such a chimera to protect against the Gram-negative Xylella fastidiosa (Xf), which causes diseases in multiple plants of economic importance. Here we report the design and delivery of this chimera to target the Xf subspecies fastidiosa (Xff), which causes Pierce disease in grapevines and poses a great threat to the wine-growing regions of California. One domain of this chimera is an elastase that recognizes and cleaves MopB, a conserved outer membrane protein of Xff. The second domain is a lytic peptide, cecropin B, which targets conserved lipid moieties and creates pores in the Xff outer membrane. A flexible linker joins the recognition and lysis domains, thereby ensuring correct folding of the individual domains and synergistic combination of their functions. The chimera transgene is fused with an amino-terminal signal sequence to facilitate delivery of the chimera to the plant xylem, the site of Xff colonization. We demonstrate that the protein chimera expressed in the xylem is able to directly target Xff, suppress its growth, and significantly decrease the leaf scorching and xylem clogging commonly associated with Pierce disease in grapevines. We believe that similar strategies involving protein chimeras can be developed to protect against many diseases caused by human and plant pathogens. PMID:22355130

  4. Temporal and spatial scaling of the genetic structure of a vector-borne plant pathogen.

    PubMed

    Coletta-Filho, Helvécio D; Francisco, Carolina S; Almeida, Rodrigo P P

    2014-02-01

    The ecology of plant pathogens of perennial crops is affected by the long-lived nature of their immobile hosts. In addition, changes to the genetic structure of pathogen populations may affect disease epidemiology and management practices; examples include local adaptation of more fit genotypes or introduction of novel genotypes from geographically distant areas via human movement of infected plant material or insect vectors. We studied the genetic structure of Xylella fastidiosa populations causing disease in sweet orange plants in Brazil at multiple scales using fast-evolving molecular markers (simple-sequence DNA repeats). Results show that populations of X. fastidiosa were regionally isolated, and that isolation was maintained for populations analyzed a decade apart from each other. However, despite such geographic isolation, local populations present in year 2000 were largely replaced by novel genotypes in 2009 but not as a result of migration. At a smaller spatial scale (individual trees), results suggest that isolates within plants originated from a shared common ancestor. In summary, new insights on the ecology of this economically important plant pathogen were obtained by sampling populations at different spatial scales and two different time points. PMID:24397266

  5. A Computerized Evaluation Methodology for Pre-Hospital EMS Cardiac Care

    PubMed Central

    Nagurney, Frank K.

    1980-01-01

    The computerized application of cardiac care protocols for pre-hospital EMS care is presented. The program logic is reviewed and an example of its application is provided. Uses of the results of the program in EMS management are suggested.

  6. EMS Stretcher “Misadventures” in a Large, Urban EMS System: A Descriptive Analysis of Contributing Factors and Resultant Injuries

    PubMed Central

    Goodloe, Jeffrey M.; Crowder, Christopher J.; Arthur, Annette O.; Thomas, Stephen H.

    2012-01-01

    Purpose. There is a paucity of data regarding EMS stretcher-operation-related injuries. This study describes and analyzes characteristics associated with undesirable stretcher operations, with or without resultant injury in a large, urban EMS agency. Methods. In the study agency, all stretcher-related “misadventures” are required to be documented, regardless of whether injury results. All stretcher-related reports between July 1, 2009 and June 30, 2010 were queried in retrospective analysis, avoiding Hawthorne effect in stretcher operations. Results. During the year studied, 129,110 patients were transported. 23 stretcher incidents were reported (0.16 per 1,000 transports). No patient injury occurred. Four EMS providers sustained minor injuries. Among contributing aspects, the most common involved operations surrounding the stretcher-ambulance safety latch, 14/23 (60.9%). From a personnel injury prevention perspective, there exists a significant relationship between combative patients and crew injury related to stretcher operation, Fisher's exact test 0.048. Conclusions. In this large, urban EMS system, the incidence of injury related to stretcher operations in the one-year study period is markedly low, with few personnel injuries and no patient injuries incurred. Safety for EMS personnel and patients could be advanced by educational initiatives that highlight specific events and conditions contributing to stretcher-related adverse events. PMID:22606379

  7. Recent technical advancements enabled atomic resolution CryoEM

    NASA Astrophysics Data System (ADS)

    Xueming, Li

    2016-01-01

    With recent breakthroughs in camera and image processing technologies single-particle electron cryo-microscopy (CryoEM) has suddenly gained the attention of structural biologists as a powerful tool able to solve the atomic structures of biological complexes and assemblies. Compared with x-ray crystallography, CryoEM can be applied to partially flexible structures in solution and without the necessity of crystallization, which is especially important for large complexes and assemblies. This review briefly explains several key bottlenecks for atomic resolution CryoEM, and describes the corresponding solutions for these bottlenecks based on the recent technical advancements. The review also aims to provide an overview about the technical differences between its applications in biology and those in material science. Project supported by Tsinghua-Peking Joint Center for Life Sciences, China.

  8. Virus particle dynamics derived from CryoEM studies.

    PubMed

    Doerschuk, Peter C; Gong, Yunye; Xu, Nan; Domitrovic, Tatiana; Johnson, John E

    2016-06-01

    The direct electron detector has revolutionized electron cryo-microscopy (CryoEM). Icosahedral virus structures are routinely produced at 4Å resolution or better and the approach has largely displaced virus crystallography, as it requires less material, less purity and often produces a structure more rapidly. Largely ignored in this new era of CryoEM is the dynamic information in the data sets that was not available in X-ray structures. Here we review an approach that captures the dynamic character of viruses displayed in the CryoEM ensemble of particles at the moment of freezing. We illustrate the approach with a simple model, briefly describe the details and provide a practical application to virus particle maturation. PMID:27085980

  9. Geographic distribution and relative abundance of the invasive glassy-winged sharpshooter: effects of temperature and egg parasitoids.

    PubMed

    Gutierrez, Andrew Paul; Ponti, Luigi; Hoddle, Mark; Almeida, Rodrigo P P; Irvin, Nicola A

    2011-08-01

    The capacity to predict the geographic distribution and relative abundance of invasive species is pivotal to developing policy for eradication or control and management. Commonly used methods fall under the ambit of ecological niche models (ENMs). These methods were reviewed and shortcomings identified. Weather-driven physiologically based demographic models (PBDMs) are proposed that resolve many of the deficiencies of ENMs. The PBDM approach is used to analyze the invasiveness of the polyphagous glassy-winged sharpshooter (Homalodisca vitripennis [Germar]), a pest native to the southeastern United States and northeastern Mexico that extended its range into California in 1989. Glassy-winged sharpshooter vectors the pathogenic bacterium, Xylella fastidiosa (Wells) that causes Pierce's disease in grape and scorch-like diseases in other plants. PBDMs for glassy-winged sharpshooter and its egg parasitoids (Gonatocerus ashmeadi Girault and G. triguttatus Girault) were developed and linked to a PBDM for grape published by Wermelinger et al. (1991). Daily weather data from 108 locations across California for the period 1995-2006 were used to drive the PBDM system, and GRASS GIS was used to map the simulation results. The geographic distribution of glassy-winged sharpshooter, as observed, is predicted to be largely restricted to the warm areas of southern California, with the action of the two egg parasitoids reducing its abundance >90%. The average indispensable mortality contributed by G. triguttatus is <1%. A temperature-dependent developmental rate model for X. fastidiosa was developed that suggests its geographic range is also limited to the warm inland areas of southern California. Biological control of glassy-winged sharpshooter further decreases the pathogen's relative range. Climate warming scenarios of +2°C and +3°C suggest that the distribution and severity of glassy-winged sharpshooter and X. fastidiosa will increase in the agriculturally rich central valley

  10. Geospatial Analysis of Pediatric EMS Run Density and Endotracheal Intubation

    PubMed Central

    Hansen, Matthew; Loker, William; Warden, Craig

    2016-01-01

    Introduction The association between geographic factors, including transport distance, and pediatric emergency medical services (EMS) run clustering on out-of-hospital pediatric endotracheal intubation is unclear. The objective of this study was to determine if endotracheal intubation procedures are more likely to occur at greater distances from the hospital and near clusters of pediatric calls. Methods This was a retrospective observational study including all EMS runs for patients less than 18 years of age from 2008 to 2014 in a geographically large and diverse Oregon county that includes densely populated urban areas near Portland and remote rural areas. We geocoded scene addresses using the automated address locator created in the cloud-based mapping platform ArcGIS, supplemented with manual address geocoding for remaining cases. We then use the Getis-Ord Gi spatial statistic feature in ArcGIS to map statistically significant spatial clusters (hot spots) of pediatric EMS runs throughout the county. We then superimposed all intubation procedures performed during the study period on maps of pediatric EMS-run hot spots, pediatric population density, fire stations, and hospitals. We also performed multivariable logistic regression to determine if distance traveled to the hospital was associated with intubation after controlling for several confounding variables. Results We identified a total of 7,797 pediatric EMS runs during the study period and 38 endotracheal intubations. In univariate analysis we found that patients who were intubated were similar to those who were not in gender and whether or not they were transported to a children’s hospital. Intubated patients tended to be transported shorter distances and were older than non-intubated patients. Increased distance from the hospital was associated with reduced odds of intubation after controlling for age, sex, scene location, and trauma system entry status in a multivariate logistic regression. The

  11. Speaker verification using combined acoustic and EM sensor signal processing

    SciTech Connect

    Ng, L C; Gable, T J; Holzrichter, J F

    2000-11-10

    Low Power EM radar-like sensors have made it possible to measure properties of the human speech production system in real-time, without acoustic interference. This greatly enhances the quality and quantity of information for many speech related applications. See Holzrichter, Burnett, Ng, and Lea, J. Acoustic. SOC. Am . 103 ( 1) 622 (1998). By combining the Glottal-EM-Sensor (GEMS) with the Acoustic-signals, we've demonstrated an almost 10 fold reduction in error rates from a speaker verification system experiment under a moderate noisy environment (-10dB).

  12. Low Bandwidth Vocoding using EM Sensor and Acoustic Signal Processing

    SciTech Connect

    Ng, L C; Holzrichter, J F; Larson, P E

    2001-10-25

    Low-power EM radar-like sensors have made it possible to measure properties of the human speech production system in real-time, without acoustic interference [1]. By combining these data with the corresponding acoustic signal, we've demonstrated an almost 10-fold bandwidth reduction in speech compression, compared to a standard 2.4 kbps LPC10 protocol used in the STU-III (Secure Terminal Unit, third generation) telephone. This paper describes a potential EM sensor/acoustic based vocoder implementation.

  13. Developments in the EM-CCD camera for OGRE

    NASA Astrophysics Data System (ADS)

    Tutt, James H.; McEntaffer, Randall L.; DeRoo, Casey; Schultz, Ted; Miles, Drew M.; Zhang, William; Murray, Neil J.; Holland, Andrew D.; Cash, Webster; Rogers, Thomas; O'Dell, Steve; Gaskin, Jessica; Kolodziejczak, Jeff; Evagora, Anthony M.; Holland, Karen; Colebrook, David

    2014-07-01

    The Off-plane Grating Rocket Experiment (OGRE) is a sub-orbital rocket payload designed to advance the development of several emerging technologies for use on space missions. The payload consists of a high resolution soft X-ray spectrometer based around an optic made from precision cut and ground, single crystal silicon mirrors, a module of off-plane gratings and a camera array based around Electron Multiplying CCD (EM-CCD) technology. This paper gives an overview of OGRE with emphasis on the detector array; specifically this paper will address the reasons that EM-CCDs are the detector of choice and the advantages and disadvantages that this technology offers.

  14. Item Parameter Estimation via Marginal Maximum Likelihood and an EM Algorithm: A Didactic.

    ERIC Educational Resources Information Center

    Harwell, Michael R.; And Others

    1988-01-01

    The Bock and Aitkin Marginal Maximum Likelihood/EM (MML/EM) approach to item parameter estimation is an alternative to the classical joint maximum likelihood procedure of item response theory. This paper provides the essential mathematical details of a MML/EM solution and shows its use in obtaining consistent item parameter estimates. (TJH)

  15. 7 CFR 1945.35 - Special EM loan training.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... Agriculture Regulations of the Department of Agriculture (Continued) RURAL HOUSING SERVICE, RURAL BUSINESS...) PROGRAM REGULATIONS (CONTINUED) EMERGENCY Disaster Assistance-General § 1945.35 Special EM loan training. (a) General. When it is evident that a large number of farmers were affected by a widespread...

  16. Reservoir characterization and steam flood monitoring with crosshole EM

    SciTech Connect

    Wilt, M.; Torres-Verdin, C.

    1995-06-01

    Crosshole electromagnetic (EM) imaging is applied to reservoir characterization and steam flood monitoring in a central California oil field. Steam was injected into three stacked eastward-dipping, unconsolidated oil sands within the upper 200 m. The steam plume is expected to develop as an ellipse aligned with the regional northwest-southeast strike. EM measurements were made from two fiberglass-cased observation wells straddling the steam injector on a northeast-southwest profile using the LLNL frequency domain crosshole EM system. Field data were collected before the initiation of a steam drive to map the distribution of the oil sands and then 6 and 12 months later to monitor the progress of the steam chest. Resistivity images derived from the EM data before steam injection clearly delineate the distribution and dipping structure on the target oil sands. Difference images, from data collected before and after steam flooding, show resistivity changes that indicate that the steam chest has developed only in the deeper oil sands although steam injection occurred in all three sand layers.

  17. 7 CFR 1945.20 - Making EM loans available.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 7 Agriculture 13 2010-01-01 2009-01-01 true Making EM loans available. 1945.20 Section 1945.20 Agriculture Regulations of the Department of Agriculture (Continued) RURAL HOUSING SERVICE, RURAL BUSINESS-COOPERATIVE SERVICE, RURAL UTILITIES SERVICE, AND FARM SERVICE AGENCY, DEPARTMENT OF AGRICULTURE...

  18. Functionalized anatomical models for EM-neuron Interaction modeling.

    PubMed

    Neufeld, Esra; Cassará, Antonino Mario; Montanaro, Hazael; Kuster, Niels; Kainz, Wolfgang

    2016-06-21

    The understanding of interactions between electromagnetic (EM) fields and nerves are crucial in contexts ranging from therapeutic neurostimulation to low frequency EM exposure safety. To properly consider the impact of in vivo induced field inhomogeneity on non-linear neuronal dynamics, coupled EM-neuronal dynamics modeling is required. For that purpose, novel functionalized computable human phantoms have been developed. Their implementation and the systematic verification of the integrated anisotropic quasi-static EM solver and neuronal dynamics modeling functionality, based on the method of manufactured solutions and numerical reference data, is described. Electric and magnetic stimulation of the ulnar and sciatic nerve were modeled to help understanding a range of controversial issues related to the magnitude and optimal determination of strength-duration (SD) time constants. The results indicate the importance of considering the stimulation-specific inhomogeneous field distributions (especially at tissue interfaces), realistic models of non-linear neuronal dynamics, very short pulses, and suitable SD extrapolation models. These results and the functionalized computable phantom will influence and support the development of safe and effective neuroprosthetic devices and novel electroceuticals. Furthermore they will assist the evaluation of existing low frequency exposure standards for the entire population under all exposure conditions. PMID:27224508

  19. Functionalized anatomical models for EM-neuron Interaction modeling

    NASA Astrophysics Data System (ADS)

    Neufeld, Esra; Cassará, Antonino Mario; Montanaro, Hazael; Kuster, Niels; Kainz, Wolfgang

    2016-06-01

    The understanding of interactions between electromagnetic (EM) fields and nerves are crucial in contexts ranging from therapeutic neurostimulation to low frequency EM exposure safety. To properly consider the impact of in vivo induced field inhomogeneity on non-linear neuronal dynamics, coupled EM-neuronal dynamics modeling is required. For that purpose, novel functionalized computable human phantoms have been developed. Their implementation and the systematic verification of the integrated anisotropic quasi-static EM solver and neuronal dynamics modeling functionality, based on the method of manufactured solutions and numerical reference data, is described. Electric and magnetic stimulation of the ulnar and sciatic nerve were modeled to help understanding a range of controversial issues related to the magnitude and optimal determination of strength-duration (SD) time constants. The results indicate the importance of considering the stimulation-specific inhomogeneous field distributions (especially at tissue interfaces), realistic models of non-linear neuronal dynamics, very short pulses, and suitable SD extrapolation models. These results and the functionalized computable phantom will influence and support the development of safe and effective neuroprosthetic devices and novel electroceuticals. Furthermore they will assist the evaluation of existing low frequency exposure standards for the entire population under all exposure conditions.

  20. Texas Hold 'em Online Poker: A Further Examination

    ERIC Educational Resources Information Center

    Hopley, Anthony A. B.; Dempsey, Kevin; Nicki, Richard

    2012-01-01

    Playing Texas Hold 'em Online Poker (THOP) is on the rise. However, there is relatively little research examining factors that contribute to problem gambling in poker players. The aim of this study was to extend the research findings of Hopley and Nicki (2010). The negative mood states of depression, anxiety and stress were found to be linked to…

  1. Airborne EM for geothermal and hydrogeological mapping

    NASA Astrophysics Data System (ADS)

    Menghini, A.; Manzella, A.; Viezzoli, A.; Montanari, D.; Maggi, S.

    2012-12-01

    Within the "VIGOR" project, aimed at assessing the geothermal potential of four regions in southern Italy, Airborne EM data have been acquired, modeled and interpreted. The system deployed was SkyTEM, a time-domain helicopter electromagnetic system designed for hydrogeophysical, environmental and mineral investigations. The AEM data provide, after data acquisition, analysis, processing, and modeling, a distribution volume of electrical resistivity, spanning an investigation depth from ground surface of few hundred meters, depending on resistivity condition. Resistivity is an important physical parameter for geothermal investigation, since it proved to be very effective in mapping anomalies due to hydrothermal fluid circulation, which usually has high salt content and produces clayey alteration minerals. Since the project required, among other issues, to define geothermal resources at shallow level, it was decided to perform a test with an airborne electromagnetic geophysical survey, to verify the advantages offered by the system in covering large areas in a short time. The geophysical survey was carried out in Sicily, Italy, in late 2011, over two test sites named "Termini" and "Western Sicily". The two areas were chosen on different basis. "Termini" area is covered by extensive geological surveys, and was going to be investigated also by means of electrical tomography in its northern part. Since geological condition of Sicily, even at shallow depth, is very complex, this area provided a good place for defining the resistivity values of the main geological units outcropping in the region. "Termini" survey has been also an occasion to define relations between resistivity distribution, lithological units and thermal conductivity. The "Western Sicily" area cover the main thermal manifestations of western Sicily, and the research target was to establish whether they are characterized by common hydrogeological or tectonic features that could be mapped by resistivity

  2. Development of the Emergency Medical Services Role Identity Scale (EMS-RIS).

    PubMed

    Donnelly, Elizabeth A; Siebert, Darcy; Siebert, Carl

    2015-01-01

    This article describes the development and validation of the theoretically grounded Emergency Medical Services Role Identity Scale (EMS-RIS), which measures four domains of EMS role identity. The EMS-RIS was developed using a mixed methods approach. Key informants informed item development and the scale was validated using a representative probability sample of EMS personnel. Factor analyses revealed a conceptually consistent, four-factor solution with sound psychometric properties as well as evidence of convergent and discriminant validities. Social workers work with EMS professionals in crisis settings and as their counselors when they are distressed. The EMS-RIS provides useful information for the assessment of and intervention with distressed EMS professionals, as well as how role identity may influence occupational stress. PMID:25760489

  3. Nonlinear Smoothing and the EM Algorithm for Positive Integral Equations of the First Kind

    SciTech Connect

    Eggermont, P. P. B.

    1999-01-15

    We study a modification of the EMS algorithm in which each step of the EMS algorithm is preceded by a nonlinear smoothing step of the form Nf-exp(S*log f) , where S is the smoothing operator of the EMS algorithm. In the context of positive integral equations (a la positron emission tomography) the resulting algorithm is related to a convex minimization problem which always admits a unique smooth solution, in contrast to the unmodified maximum likelihood setup. The new algorithm has slightly stronger monotonicity properties than the original EM algorithm. This suggests that the modified EMS algorithm is actually an EM algorithm for the modified problem. The existence of a smooth solution to the modified maximum likelihood problem and the monotonicity together imply the strong convergence of the new algorithm. We also present some simulation results for the integral equation of stereology, which suggests that the new algorithm behaves roughly like the EMS algorithm.

  4. GPS computer navigators to shorten EMS response and transport times.

    PubMed

    Ota, F S; Muramatsu, R S; Yoshida, B H; Yamamoto, L G

    2001-05-01

    GPS (global positioning satellite system to determine one's position on earth) units have become inexpensive and compact. The purpose of this study is to assess the effectiveness of a GPS enhanced computer street map navigator to improve the ability of EMS drivers in an urban setting to locate their destination and shorten response times. For part I, residential addresses in the city were randomly selected from a telephone directory. Two driver/navigator teams were assigned to drive to the address adhering to speed limits. One team used a standard street map, whereas the other team used a GPS computer navigator. The travel time and distance of the runs were compared. For part II, the computer GPS navigator was placed on an ambulance to supplement their normal methods of navigation to find the address requesting EMS. After the run was completed, EMS providers were interviewed to determine their opinion of whether the GPS navigator was helpful. For part I the results showed that in the 29 initial test runs, comparing the GPS team versus the standard map team, the mean distances traveled were 8.7 versus 9.0 kilometers (not significant) and the mean travel times were 13.5 versus 14.6 minutes (P=.02), respectively. The GPS team arrived faster in 72% runs. For part II the results showed that most EMS providers surveyed noted that the GPS computer navigator enhanced their ability to find the destination and all EMS providers acknowledged that it would enhance their ability to find a destination in an area in which they were unfamiliar. These results suggest that a portable GPS computer navigator system is helpful and can enhance the ability of prehospital care providers to locate their destination. Because these units are accurate and inexpensive, GPS computer navigators may be a valuable tool in reducing pre-hospital transport times. PMID:11326345

  5. The B and Be States of the Star EM Cepheus

    NASA Astrophysics Data System (ADS)

    Kjurkchieva, Diana; Marchev, Dragomir; Sigut, T. A. A.; Dimitrov, Dinko

    2016-09-01

    We present 11 yr of high-resolution, spectroscopic observations for the star EM Cep. EM Cep switches between B and Be star states, as revealed by the level of Hα emission, but spends most of its time in the B star state. EM Cep has been considered to be an eclipsing, near-contact binary of nearly equal-mass B stars in order to reproduce regular photometric variations; however, this model is problematic due to the lack of any observed Doppler shift in the spectrum. Our observations confirm that there are no apparent Doppler shifts in the wide spectral lines Hα and He i λ6678 in either the B or Be star states. The profiles of He i λ6678 typically exhibited a filled-in absorption core, but we detected weak emission in this line during the highest Be state. Given the lack of observed Doppler shifts, we model EM Cep as an isolated Be star with a variable circumstellar disk. We can reproduce the observed Hα emission profiles over the 11 yr period reasonably well with disk masses on the order of 3{--}10× {10}-11 {M}* in the Be state with the circumstellar disk seen at an inclination of 78° to the line of sight. From a disk ejection episode in 2014, we estimate a mass-loss rate of ≈ 3× {10}-9 {M}ȯ {{yr}}-1. The derived disk density parameters are typical of those found for the classical Be stars. We therefore suggest that the EM Cep is a classical Be star and that its photometric variations are the result of β Cep or nonradial pulsations.

  6. Similarity-regulation of OS-EM for accelerated SPECT reconstruction.

    PubMed

    Vaissier, P E B; Beekman, F J; Goorden, M C

    2016-06-01

    Ordered subsets expectation maximization (OS-EM) is widely used to accelerate image reconstruction in single photon emission computed tomography (SPECT). Speedup of OS-EM over maximum likelihood expectation maximization (ML-EM) is close to the number of subsets used. Although a high number of subsets can shorten reconstruction times significantly, it can also cause severe image artifacts such as improper erasure of reconstructed activity if projections contain few counts. We recently showed that such artifacts can be prevented by using a count-regulated OS-EM (CR-OS-EM) algorithm which automatically adapts the number of subsets for each voxel based on the estimated number of counts that the voxel contributed to the projections. While CR-OS-EM reached high speed-up over ML-EM in high-activity regions of images, speed in low-activity regions could still be very slow. In this work we propose similarity-regulated OS-EM (SR-OS-EM) as a much faster alternative to CR-OS-EM. SR-OS-EM also automatically and locally adapts the number of subsets, but it uses a different criterion for subset regulation: the number of subsets that is used for updating an individual voxel depends on how similar the reconstruction algorithm would update the estimated activity in that voxel with different subsets. Reconstructions of an image quality phantom and in vivo scans show that SR-OS-EM retains all of the favorable properties of CR-OS-EM, while reconstruction speed can be up to an order of magnitude higher in low-activity regions. Moreover our results suggest that SR-OS-EM can be operated with identical reconstruction parameters (including the number of iterations) for a wide range of count levels, which can be an additional advantage from a user perspective since users would only have to post-filter an image to present it at an appropriate noise level. PMID:27206135

  7. Similarity-regulation of OS-EM for accelerated SPECT reconstruction

    NASA Astrophysics Data System (ADS)

    Vaissier, P. E. B.; Beekman, F. J.; Goorden, M. C.

    2016-06-01

    Ordered subsets expectation maximization (OS-EM) is widely used to accelerate image reconstruction in single photon emission computed tomography (SPECT). Speedup of OS-EM over maximum likelihood expectation maximization (ML-EM) is close to the number of subsets used. Although a high number of subsets can shorten reconstruction times significantly, it can also cause severe image artifacts such as improper erasure of reconstructed activity if projections contain few counts. We recently showed that such artifacts can be prevented by using a count-regulated OS-EM (CR-OS-EM) algorithm which automatically adapts the number of subsets for each voxel based on the estimated number of counts that the voxel contributed to the projections. While CR-OS-EM reached high speed-up over ML-EM in high-activity regions of images, speed in low-activity regions could still be very slow. In this work we propose similarity-regulated OS-EM (SR-OS-EM) as a much faster alternative to CR-OS-EM. SR-OS-EM also automatically and locally adapts the number of subsets, but it uses a different criterion for subset regulation: the number of subsets that is used for updating an individual voxel depends on how similar the reconstruction algorithm would update the estimated activity in that voxel with different subsets. Reconstructions of an image quality phantom and in vivo scans show that SR-OS-EM retains all of the favorable properties of CR-OS-EM, while reconstruction speed can be up to an order of magnitude higher in low-activity regions. Moreover our results suggest that SR-OS-EM can be operated with identical reconstruction parameters (including the number of iterations) for a wide range of count levels, which can be an additional advantage from a user perspective since users would only have to post-filter an image to present it at an appropriate noise level.

  8. Service Discovery Framework Supported by EM Algorithm and Bayesian Classifier

    NASA Astrophysics Data System (ADS)

    Peng, Yanbin

    Service oriented computing has become the main stream research field nowadays. Meanwhile, machine learning is a promising AI technology which can enhance the performance of traditional algorithm. Therefore, aiming at solving service discovery problem, this paper imports Bayesian classifier to web service discovery framework, which can improve service querying speed. In this framework, services in service library become training set of Bayesian classifier, service query becomes a testing sample. Service matchmaking process can be executed in related service class, which has fewer services, thus can save time. Due to don't know the class of service in training set, EM algorithm is used to estimate prior probability and likelihood functions. Experiment results show that the EM algorithm and Bayesian classifier supported method outperforms other methods in time complexity.

  9. Magen David Adom--the EMS in Israel.

    PubMed

    Ellis, Daniel Y; Sorene, Eliot

    2008-01-01

    Israel is a small country with a population of around 7 million. The sole EMS provider for Israel is Magen David Adom (MDA) (translated as 'Red Shield of David'). MDA also carries out the functions of a National Society (similar to the Red Cross) and provides all the blood and blood product services for the country. Nationwide, the organisation responds to over 1000 emergency calls a day and uses doctors, paramedics, emergency medical technicians and volunteers. Local geopolitics has meant that MDA has to be prepared for anything from everyday emergency calls to suicide bombings and regional wars. MDA also prides itself in being able to rapidly assemble and dispatch mobile aid teams to scenes of international disasters. Such a broad range of activities is unusual for a single EMS organisation. PMID:17767990

  10. Superconducting Electromagnetic Suspension (EMS) system for Grumman Maglev concept

    NASA Technical Reports Server (NTRS)

    Kalsi, Swarn S.

    1994-01-01

    The Grumman developed Electromagnetic Suspension (EMS) Maglev system has the following key characteristics: a large operating airgap--40 mm; levitation at all speeds; both high speed and low speed applications; no deleterious effects on SC coils at low vehicle speeds; low magnetic field at the SC coil--less than 0.35 T; no need to use non-magnetic/non-metallic rebar in the guideway structure; low magnetic field in passenger cabin--approximately 1 G; low forces on the SC coil; employs state-of-the-art NbTi wire; no need for an active magnet quench protection system; and lower weight than a magnet system with copper coils. The EMS Maglev described in this paper does not require development of any new technologies. The system could be built with the existing SC magnet technology.

  11. Environmental Restoration and Waste Management (EM) program: An introduction

    SciTech Connect

    Not Available

    1990-12-01

    This booklet introduces the reader to the mission and functions of a major new unit within the US Department of Energy (DOE): the Office of Environmental Restoration and Waste Management (EM). The Secretary of Energy established EM in November 1989, implementing a central purpose of DOE's first annual Environmental Restoration and Waste Management Five-Year Plan, which had appeared three months earlier. The contents of this booklet, and their arrangement, reflect the annual update of the Five-Year Plan. The Five-Year Plan supports DOE's strategy for meeting its 30-year compliance and cleanup goal. This strategy involves: focusing DOE's activities on eliminating or reducing known or recognized potential risks to worker and public health and the environment, containing or isolating, removing, or detoxifying onsite and offsite contamination, and developing technology to achieve DOE's environmental goals.

  12. Generalized single-particle cryo-EM--a historical perspective.

    PubMed

    Frank, Joachim

    2016-02-01

    This is a brief account of the earlier history of single-particle cryo-EM of biological molecules lacking internal symmetry, which goes back to the mid-seventies. The emphasis of this review is on the mathematical concepts and computational approaches. It is written as the field experiences a turning point in the wake of the introduction of digital cameras capable of single electron counting, and near-atomic resolution can be reached even for smaller molecules. PMID:26566976

  13. EMS-STARS: Emergency Medical Services "Superuser" Transport Associations: An Adult Retrospective Study.

    PubMed

    Hall, M Kennedy; Raven, Maria C; Hall, Jane; Yeh, Clement; Allen, Elaine; Rodriguez, Robert M; Tangherlini, Niels L; Sporer, Karl A; Brown, John F

    2015-01-01

    Abstract Objective. Emergency medical services (EMS) "superusers" -those who use EMS services at extremely high rates -have not been well characterized. Recent interest in the small group of individuals who account for a disproportionate share of health-care expenditures has led to research on frequent users of emergency departments and other health services, but little research has been done regarding those who use EMS services. To inform policy and intervention implementation, we undertook a descriptive analysis of EMS superusers in a large urban community. In this paper we compare EMS superusers to low, moderate, and high users to characterize factors contributing to EMS use. We also estimate the financial impact of EMS superusers. Methods. We conducted a retrospective cross-sectional study based on 1 year of data from an urban EMS system. Data for all EMS encounters with patients age ≥18 years were extracted from electronic records generated on scene by paramedics. We identified demographic and clinical variables associated with levels of EMS use. EMS users were characterized by the annual number of EMS encounters: low (1), moderate (2-4), high (5-14), and superusers (≥15). In addition, we performed a financial analysis using San Francisco Fire Department (SFFD) 2009 charge and reimbursement data. Results. A total of 31,462 adults generated 43,559 EMS ambulance encounters, which resulted in 39,107 transports (a 90% transport rate). Encounters for general medical reasons were common among moderate and high users and less frequent among superusers and low users, while alcohol use was exponentially correlated with encounter frequency. Superusers were significantly younger than moderate EMS users, and more likely to be male. The superuser group created a significantly higher financial burden/person than any other group, comprising 0.3% of the study population, but over 6% of annual EMS charges and reimbursements. Conclusions. In this retrospective study, adult

  14. Improving EM&V for Energy Efficiency Programs (Fact Sheet)

    SciTech Connect

    Not Available

    2012-07-01

    This fact sheet describes the objectives of the U.S. Department of Energy Uniform Methods Project to bring consistency to energy savings calculations in U.S. energy efficiency programs. The U.S. Department of Energy (DOE) is developing a framework and a set of protocols for determining gross energy savings from energy efficiency measures and programs. The protocols represent a refinement of the body of knowledge supporting energy efficiency evaluation, measurement, and verification (EM&V) activities. They have been written by technical experts within the field and reviewed by industry experts. Current EM&V practice allows for multiple methods for calculating energy savings. These methods were developed to meet the needs of energy efficiency program administrators and regulators. Although they served their original objectives well, they have resulted in inconsistent and incomparable savings results - even for identical measures. The goal of the Uniform Methods Project is to strengthen the credibility of energy savings determinations by improving EM&V, increasing the consistency and transparency of how energy savings are determined.

  15. The US DOE-EM International Program - 13004

    SciTech Connect

    Elmetti, Rosa R.; Han, Ana M.; Williams, Alice C.

    2013-07-01

    The U.S. Department of Energy (DOE) Office of Environmental Management (EM) conducts international collaboration activities in support of U.S. policies and objectives regarding the accelerated risk reduction and remediation of environmental legacy of the nations' nuclear weapons program and government sponsored nuclear energy research. The EM International Program supported out of the EM Office of the Associate Principal Deputy Assistant Secretary pursues collaborations with foreign government organizations, educational institutions and private industry to assist in identifying technologies and promote international collaborations that leverage resources and link international experience and expertise. In fiscal year (FY) 2012, the International Program awarded eight international collaborative projects for work scope spanning waste processing, groundwater and soil remediation, deactivation and decommissioning (D and D) and nuclear materials disposition initiatives to seven foreign organizations. Additionally, the International Program's scope and collaboration opportunities were expanded to include technical as well as non-technical areas. This paper will present an overview of the on-going tasks awarded in FY 2012 and an update of upcoming international activities and opportunities for expansion into FY 2013 and beyond. (authors)

  16. Volta phase plate cryo-EM of the small protein complex Prx3.

    PubMed

    Khoshouei, Maryam; Radjainia, Mazdak; Phillips, Amy J; Gerrard, Juliet A; Mitra, Alok K; Plitzko, Jürgen M; Baumeister, Wolfgang; Danev, Radostin

    2016-01-01

    Cryo-EM of large, macromolecular assemblies has seen a significant increase in the numbers of high-resolution structures since the arrival of direct electron detectors. However, sub-nanometre resolution cryo-EM structures are rare compared with crystal structure depositions, particularly for relatively small particles (<400 kDa). Here we demonstrate the benefits of Volta phase plates for single-particle analysis by time-efficient cryo-EM structure determination of 257 kDa human peroxiredoxin-3 dodecamers at 4.4 Å resolution. The Volta phase plate improves the applicability of cryo-EM for small molecules and accelerates structure determination. PMID:26817416

  17. Standard errors for EM estimates in generalized linear models with random effects.

    PubMed

    Friedl, H; Kauermann, G

    2000-09-01

    A procedure is derived for computing standard errors of EM estimates in generalized linear models with random effects. Quadrature formulas are used to approximate the integrals in the EM algorithm, where two different approaches are pursued, i.e., Gauss-Hermite quadrature in the case of Gaussian random effects and nonparametric maximum likelihood estimation for an unspecified random effect distribution. An approximation of the expected Fisher information matrix is derived from an expansion of the EM estimating equations. This allows for inferential arguments based on EM estimates, as demonstrated by an example and simulations. PMID:10985213

  18. Modeling of MHD edge containment in strip casting with ELEKTRA and CaPS-EM codes

    SciTech Connect

    Chang, F. C.

    2000-01-12

    This paper presents modeling studies of magnetohydrodynamics analysis in twin-roll casting. Argonne National Laboratory (ANL) and ISPAT Inland Inc. (Inland), formerly Inland Steel Co., have worked together to develop a three-dimensional (3-D) computer model that can predict eddy currents, fluid flows, and liquid metal containment of an electromagnetic (EM) edge containment device. The model was verified by comparing predictions with experimental results of liquid metal containment and fluid flow in EM edge dams (EMDs) that were designed at Inland for twin-roll casting. This mathematical model can significantly shorten casting research on the use of EM fields for liquid metal containment and control. The model can optimize the EMD design so it is suitable for application, and minimize expensive time-consuming full-scale testing. Numerical simulation was performed by coupling a 3-D finite-element EM code (ELEKTRA) and a 3-D finite-difference fluids code (CaPS-EM) to solve heat transfer, fluid flow, and turbulence transport in a casting process that involves EM fields. ELEKTRA can predict the eddy-current distribution and the EM forces in complex geometries. CaPS-EM can model fluid flows with free surfaces. The computed 3-D magnetic fields and induced eddy currents in ELEKTRA are used as input to temperature- and flow-field computations in CaPS-EM. Results of the numerical simulation compared well with measurements obtained from both static and dynamic tests.

  19. Volta phase plate cryo-EM of the small protein complex Prx3

    NASA Astrophysics Data System (ADS)

    Khoshouei, Maryam; Radjainia, Mazdak; Phillips, Amy J.; Gerrard, Juliet A.; Mitra, Alok K.; Plitzko, Jürgen M.; Baumeister, Wolfgang; Danev, Radostin

    2016-01-01

    Cryo-EM of large, macromolecular assemblies has seen a significant increase in the numbers of high-resolution structures since the arrival of direct electron detectors. However, sub-nanometre resolution cryo-EM structures are rare compared with crystal structure depositions, particularly for relatively small particles (<400 kDa). Here we demonstrate the benefits of Volta phase plates for single-particle analysis by time-efficient cryo-EM structure determination of 257 kDa human peroxiredoxin-3 dodecamers at 4.4 Å resolution. The Volta phase plate improves the applicability of cryo-EM for small molecules and accelerates structure determination.

  20. Covariance Matrix Estimation for the Cryo-EM Heterogeneity Problem*

    PubMed Central

    Katsevich, E.; Katsevich, A.; Singer, A.

    2015-01-01

    In cryo-electron microscopy (cryo-EM), a microscope generates a top view of a sample of randomly oriented copies of a molecule. The problem of single particle reconstruction (SPR) from cryo-EM is to use the resulting set of noisy two-dimensional projection images taken at unknown directions to reconstruct the three-dimensional (3D) structure of the molecule. In some situations, the molecule under examination exhibits structural variability, which poses a fundamental challenge in SPR. The heterogeneity problem is the task of mapping the space of conformational states of a molecule. It has been previously suggested that the leading eigenvectors of the covariance matrix of the 3D molecules can be used to solve the heterogeneity problem. Estimating the covariance matrix is challenging, since only projections of the molecules are observed, but not the molecules themselves. In this paper, we formulate a general problem of covariance estimation from noisy projections of samples. This problem has intimate connections with matrix completion problems and high-dimensional principal component analysis. We propose an estimator and prove its consistency. When there are finitely many heterogeneity classes, the spectrum of the estimated covariance matrix reveals the number of classes. The estimator can be found as the solution to a certain linear system. In the cryo-EM case, the linear operator to be inverted, which we term the projection covariance transform, is an important object in covariance estimation for tomographic problems involving structural variation. Inverting it involves applying a filter akin to the ramp filter in tomography. We design a basis in which this linear operator is sparse and thus can be tractably inverted despite its large size. We demonstrate via numerical experiments on synthetic datasets the robustness of our algorithm to high levels of noise. PMID:25699132

  1. Perda de massa em ventos empoeirados de estrelas supergigantes

    NASA Astrophysics Data System (ADS)

    Vidotto, A. A.; Jatenco-Pereira, V.

    2003-08-01

    Em praticamente todas as regiões do diagrama HR, as estrelas apresentam evidências observacionais de perda de massa. Na literatura, pode-se encontrar trabalhos que tratam tanto do diagnóstico da perda de massa como da construção de modelos que visam explicá-la. O amortecimento de ondas Alfvén tem sido utilizado como mecanismo de aceleração de ventos homogêneos. Entretanto, sabe-se que os envelopes de estrelas frias contêm grãos sólidos e moléculas. Com o intuito de estudar a interação entre as ondas Alfvén e a poeira e a sua conseqüência na aceleração do vento estelar, Falceta-Gonçalves & Jatenco-Pereira (2002) desenvolveram um modelo de perda de massa para estrelas supergigantes. Neste trabalho, apresentamos um estudo do modelo acima proposto para avaliar a dependência da taxa de perda de massa com alguns parâmetros iniciais como, por exemplo, a densidade r0, o campo magnético B0, o comprimento de amortecimento da onda L0, seu fluxo f0, entre outros. Sendo assim, aumentando f0 de 10% a partir de valores de referência, vimos que aumenta consideravelmente, enquanto que um aumento de mesmo valor em r0, B0 e L0 acarreta uma diminuição em .

  2. EM-21 Retrieval Knowledge Center: Waste Retrieval Challenges

    SciTech Connect

    Fellinger, Andrew P.; Rinker, Michael W.; Berglin, Eric J.; Minichan, Richard L.; Poirier, Micheal R.; Gauglitz, Phillip A.; Martin, Bruce A.; Hatchell, Brian K.; Saldivar, Eloy; Mullen, O Dennis; Chapman, Noel F.; Wells, Beric E.; Gibbons, Peter W.

    2009-04-10

    EM-21 is the Waste Processing Division of the Office of Engineering and Technology, within the U.S. Department of Energy’s (DOE) Office of Environmental Management (EM). In August of 2008, EM-21 began an initiative to develop a Retrieval Knowledge Center (RKC) to provide the DOE, high level waste retrieval operators, and technology developers with centralized and focused location to share knowledge and expertise that will be used to address retrieval challenges across the DOE complex. The RKC is also designed to facilitate information sharing across the DOE Waste Site Complex through workshops, and a searchable database of waste retrieval technology information. The database may be used to research effective technology approaches for specific retrieval tasks and to take advantage of the lessons learned from previous operations. It is also expected to be effective for remaining current with state-of-the-art of retrieval technologies and ongoing development within the DOE Complex. To encourage collaboration of DOE sites with waste retrieval issues, the RKC team is co-led by the Savannah River National Laboratory (SRNL) and the Pacific Northwest National Laboratory (PNNL). Two RKC workshops were held in the Fall of 2008. The purpose of these workshops was to define top level waste retrieval functional areas, exchange lessons learned, and develop a path forward to support a strategic business plan focused on technology needs for retrieval. The primary participants involved in these workshops included retrieval personnel and laboratory staff that are associated with Hanford and Savannah River Sites since the majority of remaining DOE waste tanks are located at these sites. This report summarizes and documents the results of the initial RKC workshops. Technology challenges identified from these workshops and presented here are expected to be a key component to defining future RKC-directed tasks designed to facilitate tank waste retrieval solutions.

  3. Single-particle cryo-EM at crystallographic resolution

    PubMed Central

    Cheng, Yifan

    2015-01-01

    Until only a few years ago, single-particle electron cryo-microscopy (cryo-EM) was usually not the first choice for many structural biologists due to its limited resolution in the range of nanometer to subnanometer. Now, this method rivals X-ray crystallography in terms of resolution and can be used to determine atomic structures of macromolecules that are either refractory to crystallization or difficult to crystallize in specific functional states. In this review, I discuss the recent breakthroughs in both hardware and software that transformed cryo-microscopy, enabling understanding of complex biomolecules and their functions at atomic level. PMID:25910205

  4. A compulsator driven rapid-fire EM-gun

    SciTech Connect

    Pratap, S.B.; Bird, W.L.

    1984-03-01

    A compulsator-driven railgun is an attractive alternative to the homopolar generator-inductor-switch configuration, especially for repetitive duty. A conceptual design of a rapid-fire EM-gun system is presented. The generator is sized to accelerate a 0.08-kg projectile to 2 to 3 km/s at a 60 pulse-per-second repetition rate. Initial design parameters are discussed, and example current and velocity waveforms are given. The generator is discharged at the proper phase angle to provide a current zero just as the projectile exits the muzzle of the railgun.

  5. EM Modelling of RF Propagation Through Plasma Plumes

    NASA Astrophysics Data System (ADS)

    Pandolfo, L.; Bandinelli, M.; Araque Quijano, J. L.; Vecchi, G.; Pawlak, H.; Marliani, F.

    2012-05-01

    Electric propulsion is a commercially attractive solution for attitude and position control of geostationary satellites. Hall-effect ion thrusters generate a localized plasma flow in the surrounding of the satellite, whose impact on the communication system needs to be qualitatively and quantitatively assessed. An electromagnetic modelling tool has been developed and integrated into the Antenna Design Framework- ElectroMagnetic Satellite (ADF-EMS). The system is able to guide the user from the plume definition phases through plume installation and simulation. A validation activity has been carried out and the system has been applied to the plume modulation analysis of SGEO/Hispasat mission.

  6. Developing State and National Evaluation Infrastructures- Guidance for the Challenges and Opportunities of EM&V

    SciTech Connect

    Schiller, Steven R.; Goldman, Charles A.

    2011-06-24

    Evaluating the impacts and effectiveness of energy efficiency programs is likely to become increasingly important for state policymakers and program administrators given legislative mandates and regulatory goals and increasing reliance on energy efficiency as a resource. In this paper, we summarize three activities that the authors have conducted that highlight the expanded role of evaluation, measurement and verification (EM&V): a study that identified and analyzed challenges in improving and scaling up EM&V activities; a scoping study that identified issues involved in developing a national efficiency EM&V standard; and lessons learned from providing technical assistance on EM&V issues to states that are ramping up energy efficiency programs. The lessons learned are summarized in 13 EM&V issues that policy makers should address in each jurisdiction and which are listed and briefly described. The paper also discusses how improving the effectiveness and reliability of EM&V will require additional capacity building, better access to existing EM&V resources, new methods to address emerging issues and technologies, and perhaps foundational documents and approaches to improving the credibility and cross jurisdictional comparability of efficiency investments. Two of the potential foundational documents discussed are a national EM&V standard or resource guide and regional deemed savings and algorithm databases.

  7. EMS Instructor Training Program. National Standard Curriculum. Instructor Guide. Student Guide.

    ERIC Educational Resources Information Center

    National Highway Traffic Safety Administration (DOT), Washington, DC.

    This guide for teaching a course to prepare emergency medical service (EMS) trainers focuses on the skills necessary to present any of the Department of Transportation (DOT), National Highway Traffic Safety Administration (NHTSA) EMS courses. Course topics are as follows: (1) introduction; (2) instructor roles and responsibilities; (3) legal…

  8. Method for evaluating compatibility of commercial electromagnetic (EM) microsensor tracking systems with surgical and imaging tables

    NASA Astrophysics Data System (ADS)

    Nafis, Christopher; Jensen, Vern; von Jako, Ron

    2008-03-01

    Electromagnetic (EM) tracking systems have been successfully used for Surgical Navigation in ENT, cranial, and spine applications for several years. Catheter sized micro EM sensors have also been used in tightly controlled cardiac mapping and pulmonary applications. EM systems have the benefit over optical navigation systems of not requiring a line-of-sight between devices. Ferrous metals or conductive materials that are transient within the EM working volume may impact tracking performance. Effective methods for detecting and reporting EM field distortions are generally well known. Distortion compensation can be achieved for objects that have a static spatial relationship to a tracking sensor. New commercially available micro EM tracking systems offer opportunities for expanded image-guided navigation procedures. It is important to know and understand how well these systems perform with different surgical tables and ancillary equipment. By their design and intended use, micro EM sensors will be located at the distal tip of tracked devices and therefore be in closer proximity to the tables. Our goal was to define a simple and portable process that could be used to estimate the EM tracker accuracy, and to vet a large number of popular general surgery and imaging tables that are used in the United States and abroad.

  9. Covariance Structure Model Fit Testing under Missing Data: An Application of the Supplemented EM Algorithm

    ERIC Educational Resources Information Center

    Cai, Li; Lee, Taehun

    2009-01-01

    We apply the Supplemented EM algorithm (Meng & Rubin, 1991) to address a chronic problem with the "two-stage" fitting of covariance structure models in the presence of ignorable missing data: the lack of an asymptotically chi-square distributed goodness-of-fit statistic. We show that the Supplemented EM algorithm provides a convenient…

  10. The Relationship between the Bock-Aitkin Procedure and the EM Algorithm for IRT Model Estimation.

    ERIC Educational Resources Information Center

    Hsu, Yaowen; Ackerman, Terry A.; Fan, Meichu

    It has previously been shown that the Bock-Aitkin procedure (R. Bock and M. Aitkin, 1981) is an instance of the EM algorithm when trying to find the marginal maximum likelihood estimate for a discrete latent ability variable (latent trait). In this paper, it is shown that the Bock-Aitkin procedure is a numerical implementation of the EM algorithm…

  11. Global Convergence of the EM Algorithm for Unconstrained Latent Variable Models with Categorical Indicators

    ERIC Educational Resources Information Center

    Weissman, Alexander

    2013-01-01

    Convergence of the expectation-maximization (EM) algorithm to a global optimum of the marginal log likelihood function for unconstrained latent variable models with categorical indicators is presented. The sufficient conditions under which global convergence of the EM algorithm is attainable are provided in an information-theoretic context by…

  12. Signaling of cell fate determination by the TPD1 small protein and EMS1 receptor kinase

    PubMed Central

    Jia, Gengxiang; Liu, Xiaodong; Owen, Heather A.; Zhao, Dazhong

    2008-01-01

    Sexual reproduction requires the specification of cells with distinct fates in plants and animals. The EMS1 (also known as EXS) leucine-rich repeat receptor-like kinase (LRR-RLK) and TPD1 small protein play key roles in regulating somatic and reproductive cell fate determination in Arabidopsis anthers. Here, we show that ectopic expression of TPD1 causes abnormal differentiation of somatic and reproductive cells in anthers. In addition, ectopic TPD1 activity requires functional EMS1. Yeast two-hybrid, pull-down, and coimmunoprecipitation analyses further demonstrate that TPD1 interacts with EMS1 in vitro and in vivo. Moreover, TPD1 induces EMS1 phosphorylation in planta. Thus, our results suggest that TPD1 serves as a ligand for the EMS1 receptor kinase to signal cell fate determination during plant sexual reproduction. PMID:18250314

  13. A Generalized Fast Frequency Sweep Algorithm for Coupled Circuit-EM Simulations

    SciTech Connect

    Rockway, J D; Champagne, N J; Sharpe, R M; Fasenfest, B

    2004-01-14

    Frequency domain techniques are popular for analyzing electromagnetics (EM) and coupled circuit-EM problems. These techniques, such as the method of moments (MoM) and the finite element method (FEM), are used to determine the response of the EM portion of the problem at a single frequency. Since only one frequency is solved at a time, it may take a long time to calculate the parameters for wideband devices. In this paper, a fast frequency sweep based on the Asymptotic Wave Expansion (AWE) method is developed and applied to generalized mixed circuit-EM problems. The AWE method, which was originally developed for lumped-load circuit simulations, has recently been shown to be effective at quasi-static and low frequency full-wave simulations. Here it is applied to a full-wave MoM solver, capable of solving for metals, dielectrics, and coupled circuit-EM problems.

  14. AVALIAÇÃO DA PRESENÇA DE ENDOSSIMBIONTES Cardinium em DIFERENTES ESPÉCIES DE ARTRÓPODES.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A presença de endossimbiontes do gênero Cardinium em alguns grupos de artrópodes foi recentemente relatada e relacionada com diversas alterações reprodutivas em seus hospedeiros, tais como feminilização de ácaros, partenogênese em parasitóides, incompatibilidade citoplasmática e aumento da fecundida...

  15. Aristarchus's <em>On the Sizes and Distances of the Sun and the Moonem>: Greek and Arabic Texts

    NASA Astrophysics Data System (ADS)

    Berggren, J. L.; Sidoli, N.

    2007-05-01

    In the 1920s, T. L. Heath pointed out that historians of mathematics have "given too little attention to Aristarchus". This is still true today. The Greek text of Aristarchus's On the Sizes and Distances of the Sun and the Moonem> has received little attention; the Arabic editions virtually none. Much of what this text has to tell us about ancient and medieval mathematics and the mathematical sciences has gone unnoticed. It should be taken as an important source for our understanding of the mathematical sciences of the early Hellenistic period.

  16. The EM fields in the Solid Generated by a Fault in a Porous Region

    NASA Astrophysics Data System (ADS)

    Ren, H.; Huang, Q.; Chen, X.

    2015-12-01

    Electrokinetic effect, as one of the most possible generation mechanisms of the seismo-electromagnetic phenomenons associated with natural earthquakes, has interested many researchers. Besides, it is also considered as a potential tool for the water/oil exploration. Recently, we numerically investigated the electromagnetic (EM) fields due to the electrokinetic effect in mixed layered model. The mixed model comprises not only porous layers but also solid layers. We firstly tested a two-layer mixed model. The numerical results show that, in addition to the radiation EM fields, another kind of evanescent EM fields can be generated by the seismic waves arriving at the interface with incident angles greater the critical angle. The evanescent EM fields decay faster than the radiation EM fields when getting away from the interface. For the seismic frequency band, the evanescent EM fields in the solid are still measurable at a distance of, e.g., 2km to the interface. We then tested a eight-layer mixed model. The top and bottom layers are solid and the other layers are porous. A finite fault of 20x10km is located in the porous region. The focal depth is 8km. The applied source time function is a ramp fuction with an arise time of 0.8s. Point stacking method was used to compute the wave-fields caused by the finite fault. Our nuemrical results show that, this model can generate the EM fields before the arrival of seismic waves as well as the residual EM fields. Both the two kinds of EM fields have been observed in field observations. There is a possibility that the anomalous EM activities before big earthquakes may be caused by the fluid flow in the shallow Earth as a result of the stress changes.

  17. Review of selected oceanic EM/EO scattering problems

    NASA Astrophysics Data System (ADS)

    Haller, Merrick C.

    2010-02-01

    Electromagnetic and electro-optical (EM/EO) propagation and scattering in the ocean is of interest for a wide range of science problems. For example, the biological productivity of ocean waters through photochemical processes is governed by the vertical attenuation of solar radiation. Also, EO scattering theory is the primary basis for determining biogeochemical parameters (e.g. phytoplankton, suspended sediments, and dissolved matter) from the water leaving optical radiance. In addition, EO scattering from suspended sediments and bubbles is the limiting factor for active lidar systems used to map the sea bottom. This work will review specific applications of EO/EM scattering theory with regard to the influence of bubbles and droplets on remote sensing in the nearshore ocean. The current state of understanding concerning models and applications for optical scattering from bubbles in the water column as well as microwave scattering from water droplets produced by breaking waves at the ocean surface will be discussed as well as future research directions.

  18. The Search for Gravitational Wave EM Counterparts with Swift

    NASA Astrophysics Data System (ADS)

    Kennea, Jamie; Evans, Phil; Swift GW follow-up Team

    2016-04-01

    We present the plan to search for electromagnetic counterparts of Gravitational Waves (GWs) discovered during the current and upcoming runs of the LIGO and Virgo detectors. As we enter a period where the sensitivity of the current generation of GW detectors approaches a high probability of the first detection of a real GW signal, confirmation of the reality of these triggers will be greatly improved if an EM counterpart can be found. Swift’s ability to rapidly respond to high priority target-of-opportunity observations, it’s multi-wavelength capabilities and low overhead observing make it a seemingly ideal follow-up facility. However comparing the size of the expected GW error regions with the fields of view of the Swift XRT and UVOT telescopes, we find that covering the large GW error regions would require a unreasonably large number of pointings. We present our method of meeting this challenge, by both reducing the problem using Galaxy targeting, and by operating Swift in an entirely new way in order to cover the still large number of fields needed to chase down the EM counterpart before it disappears.

  19. Chapter 19: HVAC Controls (DDC/EMS/BAS) Evaluation Protocol

    SciTech Connect

    Romberger, J.

    2014-11-01

    The HVAC Controls Evaluation Protocol is designed to address evaluation issues for direct digital controls/energy management systems/building automation systems (DDC/EMS/BAS) that are installed to control heating, ventilation, and air-conditioning (HVAC) equipment in commercial and institutional buildings. (This chapter refers to the DDC/EMS/BAS measure as HVAC controls.) This protocol may also be applicable to industrial facilities such as clean rooms and labs, which have either significant HVAC equipment or spaces requiring special environmental conditions. This protocol addresses only HVAC-related equipment and the energy savings estimation methods associated with installing such control systems as an energy efficiency measure. The affected equipment includes: Air-side equipment (air handlers, direct expansion systems, furnaces, other heating- and cooling-related devices, terminal air distribution equipment, and fans); Central plant equipment (chillers, cooling towers, boilers, and pumps). These controls may also operate or affect other end uses, such as lighting, domestic hot water, irrigation systems, and life safety systems such as fire alarms and other security systems. Considerable nonenergy benefits, such as maintenance scheduling, system component troubleshooting, equipment failure alarms, and increased equipment lifetime, may also be associated with these systems. When connected to building utility meters, these systems can also be valuable demand-limiting control tools. However, this protocol does not evaluate any of these additional capabilities and benefits.

  20. Contactless ultrasonic treatment of melts using EM induction

    NASA Astrophysics Data System (ADS)

    Bojarevics, V.; Djambazov, G.; Lebon, G. S. B.; Pericleous, K. A.

    2015-06-01

    Ultrasound Treatment (UT) is commonly used in light alloys during solidification to refine microstructure, or disperse immersed particles. A sonotrode probe introduced into the melt generates sound waves that are strong enough to produce cavitation of dissolved gases. The same method cannot be used in high temperature melts, or for highly reactive alloys, due to probe erosion and melt contamination. An alternative, contactless method of generating sound waves is proposed and investigated theoretically in this paper, using electromagnetic (EM) induction. In addition to strong vibration, the EM induction currents generate strong stirring in the melt that aids distribution of the UT effect to large volumes of material. In a typical application, the same induction coil surrounding the crucible used to melt the alloy may be adopted for UT with suitable frequency tuning. Alternatively - or in addition - a top coil may be used. For industrial use, instead of multiple sonotrodes as has been the practice in scaling up, modelling shows that one simply has to alter the coil geometry and current to suit. To reach sinusoidal pressure fluctuations suitable for cavitation it may be necessary to tune the induction coil frequency for resonance, given the crucible dimensions.