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avrpto enhances growth and necrosis caused by pseudomonas syringae pv.tomato in tomato lines lacking either pto or prf.avrpto was introduced into three tomato genotypes with two biotic agents to study its role in compatible interactions. avrpto enhanced the capacity of the pseudomonas syringae pv. tomato strain t1 to induce necrotic symptoms on tomato plants that lacked either pto or prf genes. the enhanced necrosis correlated with a small increase in bacterial growth. in planta expression of avrpto in isolation did not elicit necrosis in the absence of a functional prf gene.200010796023
molecular and biological analysis of eight genetic islands that distinguish neisseria meningitidis from the closely related pathogen neisseria gonorrhoeae.the pathogenic species neisseria meningitidis and neisseria gonorrhoeae cause dramatically different diseases despite strong relatedness at the genetic and biochemical levels. n. meningitidis can cross the blood-brain barrier to cause meningitis and has a propensity for toxic septicemia unlike n. gonorrhoeae. we previously used subtractive hybridization to identify dna sequences which might encode functions specific to bacteremia and invasion of the meninges because they are specific to n. menin ...200010722605
mechanism of inactivation of ornithine transcarbamoylase by ndelta -(n'-sulfodiaminophosphinyl)-l-ornithine, a true transition state analogue? crystal structure and implications for catalytic mechanism.the crystal structure is reported at 1.8 a resolution of escherichia coli ornithine transcarbamoylase in complex with the active derivative of phaseolotoxin from pseudomonas syringae pv. phaseolicola, n(delta)-(n'-sulfodiaminophosphinyl)-l-ornithine. electron density reveals that the complex is not a covalent adduct as previously thought. kinetic data confirm that n(delta)-(n'-sulfodiaminophosphinyl)-l-ornithine exhibits reversible inhibition with a half-life in the order of approximately 22 h a ...200010747936
differential regulation of plastidial and cytosolic isoforms of peptide methionine sulfoxide reductase in arabidopsis.we report the characterization of two members of a gene family from arabidopsis that encode, respectively, cytosolic (cpmsr) and plastid-targeted (ppmsr) isoforms of the oxidative-stress-repair enzyme peptide methionine sulfoxide reductase. overexpression of these proteins in escherichia coli confirmed that each had pmsr enzyme activity with a synthetic substrate, n-acetyl-[(3)h]-methionine sulfoxide, or a biological substrate, alpha-1 proteinase inhibitor. the ppmsr was imported into intact chl ...200010806242
cloning and expression of ntnd, encoding a novel nad(p)(+)-independent 4-nitrobenzyl alcohol dehydrogenase from pseudomonas sp. strain tw3.pseudomonas sp. strain tw3 is able to metabolize 4-nitrotoluene to 4-nitrobenzoate and toluene to benzoate aerobically via a route analogous to the upper pathway of the tol plasmids. we report the cloning and characterization of a benzyl alcohol dehydrogenase gene (ntnd) which encodes the enzyme for the catabolism of 4-nitrobenzyl alcohol and benzyl alcohol to 4-nitrobenzaldehyde and benzaldehyde, respectively. the gene is located downstream of the previously reported ntn gene cluster. ntnd bear ...200010809692
helicobacter pylori uptake and efflux: basis for intrinsic susceptibility to antibiotics in vitro.we previously demonstrated (m. m. exner, p. doig, t. j. trust, and r. e. w. hancock, infect. immun. 63:1567-1572, 1995) that helicobacter pylori has at least one nonspecific porin, hope, which has a low abundance in the outer membrane but forms large channels. h. pylori is relatively susceptible to most antimicrobial agents but less susceptible to the polycationic antibiotic polymyxin b. we demonstrate here that h. pylori is able to take up higher basal levels of the hydrophobic fluorescent prob ...200010639345
arabidopsis cytochrome p450s that catalyze the first step of tryptophan-dependent indole-3-acetic acid biosynthesis.plants synthesize numerous secondary metabolites that are used as developmental signals or as defense against pathogens. tryptophan (trp)-derived secondary metabolites include camalexin, indole glucosinolates, and indole-3-acetic acid (iaa); however, the steps in their synthesis from trp or its precursors remain unclear. we have identified two arabidopsis cytochrome p450s (cyp79b2 and cyp79b3) that can convert trp to indole-3-acetaldoxime (iaox), a precursor to iaa and indole glucosinolates.200010681464
pilus formation and protein secretion by the same machinery in escherichia coli.the secreton (type ii secretion) and type iv pilus biogenesis branches of the general secretory pathway in gram-negative bacteria share many features that suggest a common evolutionary origin. five components of the secreton, the pseudopilins, are similar to subunits of type iv pili. here, we report that when the 15 genes encoding the pullulanase secreton of klebsiella oxytoca were expressed on a high copy number plasmid in escherichia coli, one pseudopilin, pulg, was assembled into pilus-like b ...200010811613
the gene coding for the hrp pilus structural protein is required for type iii secretion of hrp and avr proteins in pseudomonas syringae pv. tomato.bacterial surface appendages called pili often are associated with dna and/or protein transfer between cells. the exact function of pili in the transfer process is not understood and is a matter of considerable debate. the hrp pilus is assembled by the hrp type iii protein secretion system of pseudomonas syringae pv. tomato (pst) strain dc3000. in this study, we show that the hrpa gene, which encodes the major subunit of the hrp pilus, is required for secretion of putative virulence proteins, su ...200010681465
salicylic acid mediated by the oxidative burst is a key molecule in local and systemic responses of cotton challenged by an avirulent race of xanthomonas campestris pv malvacearum.we analyzed the production of reactive oxygen species, the accumulation of salicylic acid (sa), and peroxidase activity during the incompatible interaction between cotyledons of the cotton (gossypium hirsutum) cv reba b50/xanthomonas campestris pv malvacearum (xcm) race 18. sa was detected in petioles of cotyledons 6 h after infection and 24 h post inoculation in cotyledons and untreated leaves. the first peak of sa occurred 3 h after generation of superoxide (o(2)(.-)), and was inhibited by inf ...200010712539
haloalkane-utilizing rhodococcus strains isolated from geographically distinct locations possess a highly conserved gene cluster encoding haloalkane catabolism.the sequences of the 16s rrna and haloalkane dehalogenase (dhaa) genes of five gram-positive haloalkane-utilizing bacteria isolated from contaminated sites in europe, japan, and the united states and of the archetypal haloalkane-degrading bacterium rhodococcus sp. strain ncimb13064 were compared. the 16s rrna gene sequences showed less than 1% sequence divergence, and all haloalkane degraders clearly belonged to the genus rhodococcus. all strains shared a completely conserved dhaa gene, suggesti ...200010781539
a cluster of mutations disrupt the avirulence but not the virulence function of avrpto.avrpto in pseudomonas syringae pv. tomato encodes an avirulence protein that triggers race-specific resistance in tomato plants carrying pto. the avrpto protein is secreted from p. syringae pv. tomato to plant cells through the type iii secretion pathway and activates race-specific resistance by a direct interaction with the pto protein. here we report that avrpto enhances the virulence of p. syringae pv. tomato in a strain-dependent manner in tomato plants lacking pto. to determine whether the ...200010830258
purification and characterization of a catalase from the facultatively psychrophilic bacterium vibrio rumoiensis s-1(t) exhibiting high catalase activity.catalase from the facultatively psychrophilic bacterium vibrio rumoiensis s-1(t), which was isolated from an environment exposed to h(2)o(2) and exhibited high catalase activity, was purified and characterized, and its localization in the cell was determined. its molecular mass was 230 kda, and the molecule consisted of four identical subunits. the enzyme, which was not apparently reduced by dithionite, showed a soret peak at 406 nm in a resting state. the catalytic activity was 527,500 u. mg of ...200010714995
methods for intense aeration, growth, storage, and replication of bacterial strains in microtiter plates.miniaturized growth systems for heterogeneous culture collections are not only attractive in reducing demands for incubation space and medium but also in making the parallel handling of large numbers of strains more practicable. we report here on the optimization of oxygen transfer rates in deep-well microtiter plates and the development of a replication system allowing the simultaneous and reproducible sampling of 96 frozen glycerol stock cultures while the remaining culture volume remains froz ...200010831450
detection of pseudomonas savastanoi pv. savastanoi in olive plants by enrichment and pcr.the sequence of the gene iaal of pseudomonas savastanoi ew2009 was used to design primers for pcr amplification. the iaal-derived primers directed the amplification of a 454-bp fragment from genomic dna isolated from 70 strains of p. savastanoi, whereas genomic dna from 93 non-p. savastanoi isolates did not yield this amplified product. a previous bacterial enrichment in the semiselective liquid medium pvf-1 improved the pcr sensitivity level, allowing detection of 10 to 100 cfu/ml of plant extr ...200010831456
arabidopsis rela/spot homologs implicate (p)ppgpp in plant signaling.arabidopsis rpp5 is a member of a large class of pathogen resistance genes encoding nucleotide-binding sites and leucine-rich repeat domains. yeast two-hybrid analysis showed that rpp5 specifically interacts with at-rsh1, an arabidopsis rela/spot homolog. in escherichia coli, rela and spot determine the level of guanosine tetraphosphate (ppgpp) and guanosine pentaphosphate (pppgpp), which are the effector nucleotides of the bacterial stringent response. functional analysis in e. coli and in stre ...200010725385
the arcb sensor kinase of escherichia coli: genetic exploration of the transmembrane region.the arc two-component signal transduction system of escherichia coli regulates the expression of numerous operons in response to respiratory growth conditions. cellular redox state or proton motive force (delta(h(+))) has been proposed to be the signal for the membrane-associated arcb sensor kinase. this study provided evidence for a short arcb periplasmic bridge that contains a his47. the dispensability of this amino acid, the only amino acid with a pk in the physiological range, renders the de ...200010781568
inorganic polyphosphate is needed for swimming, swarming, and twitching motilities of pseudomonas aeruginosa.polyphosphate kinase (ppk), encoded by the ppk gene, is the principal enzyme in many bacteria for the synthesis of inorganic polyphosphate (poly p) from atp. a knockout mutant in the ppk gene of pseudomonas aeruginosa pao1 is impaired in flagellar swimming motility on semisolid agar plates. the mutant is deficient in type iv pili-mediated twitching motility and in a "swarming motility" previously unobserved in p. aeruginosa. in swarming cultures, the polar monotrichous bacteria have differentiat ...200010758151
identification of sinorhizobium meliloti genes regulated during symbiosis.rna fingerprinting by arbitrarily primed pcr was used to isolate sinorhizobium meliloti genes regulated during the symbiotic interaction with alfalfa (medicago sativa). sixteen partial cdnas were isolated whose corresponding genes were differentially expressed between symbiotic and free-living conditions. thirteen sequences corresponded to genes up-regulated during symbiosis, whereas three were instead repressed during establishment of the symbiotic interaction. seven cdnas corresponded to known ...200010850975
genotypic characterization of xanthomonad strains isolated from passion fruit plants (passiflora spp.) and their relatedness to different xanthomonas species.the genetic diversity of 55 xanthomonad strains isolated from passion fruit plants (passiflora spp.) and identified as xanthomonas campestris pv. passiflorae was initially assessed by randomly amplified polymorphic dna (rapd) analysis. the strains showed a high level of polymorphism with almost unique fingerprints. fifteen clusters with a similarity of approximately 70% were identified, three of which were prevalent. there was a correlation between the clusters and the geographic origin of the s ...200010758892
structure of the o polysaccharide and serological classification of pseudomonas syringae pv. ribicola ncppb 1010.the o polysaccharide (ops) moiety of the lipopolysaccharide (lps) of a phytopathogenic bacterium pseudomonas syringae pv. ribicola ncppb 1010 was studied by sugar and methylation analyses, smith degradation, and 1h- and 13c-nmr spectroscopy, including 2d cosy, tocsy, noesy and h-detected 1h,13c hmqc experiments. the ops structure was elucidated, and shown to be composed of branched pentasaccharide repeating units (o repeats) of two types, major (1) and minor (2), differing in the position of sub ...200010759863
phosphorelay as the sole physiological route of signal transmission by the arc two-component system of escherichia coli.the arc two-component system, comprising a tripartite sensor kinase (arcb) and a response regulator (arca), modulates the expression of numerous genes involved in respiratory functions. in this study, the steps of phosphoryl group transfer from phosphorylated arcb to arca were examined in vivo by using single copies of wild-type and mutant arcb alleles. the results indicate that the signal transmission occurs solely by his-asp-his-asp phosphorelay.200010851007
leucine aminopeptidases: the ubiquity of lap-n and the specificity of lap-a.the wound-induced leucine aminopeptidase (ec 3.4.11.1) genes, lapa1 and lapa2, from tomato (lycopersicon esculentum mill.) were isolated and characterized. the genes were organized in a tandem array with approximately 6 kb separating their coding regions. quantitation of lapa mrna levels in conjunction with nuclear run-on experiments indicated that lapa genes were primarily under transcriptional control after wounding and infection with pseudomonas syringae pv. tomato. in contrast, actin genes w ...200010787049
development of a highly sensitive nested-pcr procedure using a single closed tube for detection of erwinia amylovora in asymptomatic plant material.a novel method, which involves a nested pcr in a single closed tube, was developed for the sensitive detection of erwinia amylovora in plant material. the external and internal primer pairs used had different annealing temperatures and directed the amplification of a specific dna fragment from plasmid pea29. the procedure involved two consecutive pcrs, the first of which was performed at a higher annealing temperature that allowed amplification only by the external primer pair. using pure cultur ...200010788384
the gacs sensor kinase regulates alginate and poly-beta-hydroxybutyrate production in azotobacter vinelandii.azotobacter vinelandii produces two polymers: the extracellular polysaccharide alginate and the intracellular polyester poly-beta-hydroxybutyrate (phb). a cosmid clone (psmu588) from an a. vinelandii gene library diminished alginate production by a. vinelandii mucoid strain atcc 9046. the nucleotide sequence and predicted amino acid sequence of the locus responsible for the mucoidy suppression revealed 65% identity to pseudomonas gacs, a transmembrane sensor kinase of the two-component regulator ...200010762268
syringomycin e inhibition of saccharomyces cerevisiae: requirement for biosynthesis of sphingolipids with very-long-chain fatty acids and mannose- and phosphoinositol-containing head groups.syringomycin e is an antifungal cyclic lipodepsinonapeptide that inhibits the growth of saccharomyces cerevisiae by interaction with the plasma membrane. a screen conducted to find the yeast genes necessary for its fungicidal action identified two novel syringomycin e response genes, syr3 and syr4. a syr3 mutant allele was complemented by elo2 and elo3. these genes encode enzymes that catalyze the elongation of sphingolipid very long chain fatty acids. tetrad analysis showed that syr3 was elo2. ...200010770748
relationship between the tsh autotransporter and pathogenicity of avian escherichia coli and localization and analysis of the tsh genetic region.the temperature-sensitive hemagglutinin tsh is a member of the autotransporter group of proteins and was first identified in avian-pathogenic escherichia coli (apec) strain chi7122. the prevalence of tsh was investigated in 300 e. coli isolates of avian origin and characterized for virulence in a 1-day-old chick lethality test. results indicate that among the tsh-positive apec isolates, 90.6% belonged to the highest virulence class. experimental inoculation of chickens with chi7122 and an isogen ...200010858231
cytochrome p450-dependent metabolism of oxylipins in tomato. cloning and expression of allene oxide synthase and fatty acid hydroperoxide lyase.allene oxide synthase (aos) and fatty acid hydroperoxide lyase (hpl) are plant-specific cytochrome p450s that commit fatty acid hydroperoxides to different branches of oxylipin metabolism. here we report the cloning and characterization of aos (leaos) and hpl (lehpl) cdnas from tomato (lycopersicon esculentum). functional expression of the cdnas in escherichia coli showed that leaos and lehpl encode enzymes that metabolize 13- but not 9-hydroperoxide derivatives of c(18) fatty acids. leaos was a ...200010859201
the pseudomonas syringae hrp pathogenicity island has a tripartite mosaic structure composed of a cluster of type iii secretion genes bounded by exchangeable effector and conserved effector loci that contribute to parasitic fitness and pathogenicity in plants.the plant pathogenic bacterium pseudomonas syringae is divided into pathovars differing in host specificity, with p. syringae pv. syringae (psy) and p. syringae pv. tomato (pto) representing particularly divergent pathovars. p. syringae hrp/hrc genes encode a type iii protein secretion system that appears to translocate avr and hop effector proteins into plant cells. dna sequence analysis of the hrp/hrc regions in psy 61, psy b728a, and pto dc3000 has revealed a hrp pathogenicity island (pai) wi ...200010781092
functional analysis of pvds, an iron starvation sigma factor of pseudomonas aeruginosa.in pseudomonas aeruginosa, iron modulates gene expression through a cascade of negative and positive regulatory proteins. the master regulator fur is involved in iron-dependent repression of several genes. one of these genes, pvds, was predicted to encode a putative sigma factor responsible for the transcription of a subset of genes of the fur regulon. pvds appears to belong to a structurally and functionally distinct subgroup of the extracytoplasmic function family of alternative sigma factors. ...200010692351
lon protease influences antibiotic production and uv tolerance of pseudomonas fluorescens pf-5.pseudomonas fluorescens pf-5 is a soil bacterium that suppresses plant pathogens due in part to its production of the antibiotic pyoluteorin. previous characterization of pf-5 revealed three global regulators, including the stationary-phase sigma factor sigma(s) and the two-component regulators gaca and gacs, that influence both antibiotic production and stress response. in this report, we describe the serine protease lon as a fourth global regulator influencing these phenotypes in pf-5. lon mut ...200010877760
atbi-1, a plant homologue of bax inhibitor-1, suppresses bax-induced cell death in yeast and is rapidly upregulated during wounding and pathogen challenge.extensive searches have so far failed to identify functional plant homologues of the mammalian apoptotic machinery. here we report the isolation and characterisation of an arabidopsis thaliana homologue of human bax inhibitor-1, atbi-1, isolated during a differential screen of plants challenged with the phytopathogen pseudomonas syringae. atbi is a member of a small gene family in arabidopsis, members of which display extensive amino acid identity to human bi-1. atbi-1 is also functionally simil ...200010758491
characterization of an atrazine-degrading pseudaminobacter sp. isolated from canadian and french agricultural soils.atrazine, a herbicide widely used in corn production, is a frequently detected groundwater contaminant. fourteen bacterial strains able to use this herbicide as a sole source of nitrogen were isolated from soils obtained from two farms in canada and two farms in france. these strains were indistinguishable from each other based on repetitive extragenic palindromic pcr genomic fingerprinting performed with primers eric1r, eric2, and boxa1r. based on 16s rrna sequence analysis of one representativ ...200010877767
identification of the pgmg gene, encoding a bifunctional protein with phosphoglucomutase and phosphomannomutase activities, in the gellan gum-producing strain sphingomonas paucimobilis atcc 31461.the pgmg gene of sphingomonas paucimobilis atcc 31461, the industrial gellan gum-producing strain, was cloned and sequenced. it encodes a 50,059-da polypeptide that has phosphoglucomutase (pgm) and phosphomannomutase (pmm) activities and is 37 to 59% identical to other bifunctional proteins with pgm and pmm activities from gram-negative species, including pseudomonas aeruginosa algc. purified pgmg protein showed a marked preference for glucose-1-phosphate (g1p); the catalytic efficiency was abou ...200010788412
plant species and plant incubation conditions influence variability in epiphytic bacterial population size.the influences of plant species and plant incubation conditions on the variability in bacterial population sizes among leaves were investigated in field and growth chamber studies. pseudomonas syringae strains tlp2 and cit7 were inoculated onto plants and population sizes were measured at intervals after inoculation. total bacterial population sizes were also assessed in field studies. levels of leaf-to-leaf variability in both p. syringae population size and bacterial community size differed si ...200010790512
the common nodulation genes of astragalus sinicus rhizobia are conserved despite chromosomal diversity.the nodulation genes of mesorhizobium sp. (astragalus sinicus) strain 7653r were cloned by functional complementation of sinorhizobium meliloti nod mutants. the common nod genes, nodd, noda, and nodbc, were identified by heterologous hybridization and sequence analysis. the noda gene was found to be separated from nodbc by approximately 22 kb and was divergently transcribed. the 2. 0-kb noddbc region was amplified by pcr from 24 rhizobial strains nodulating a. sinicus, which represented differen ...200010877796
thr38 and ser198 are pto autophosphorylation sites required for the avrpto-pto-mediated hypersensitive response.the tomato pto kinase confers resistance to pseudomonas syringae pv. tomato expressing the avrpto protein. to elucidate the role of pto autophosphorylation in disease resistance, eight sites autophosphorylated by pto in vitro were identified by a combination of hplc purification of tryptic phosphopeptides, maldi-tof/ms analysis and edman degradation. mutational analysis of the autophosphorylation sites revealed that pto residues thr38 and ser198 are required for avrpto-pto- mediated elicitation ...200010811617
female reproductive tissues are the primary target of agrobacterium-mediated transformation by the arabidopsis floral-dip method.the floral-dip method for agrobacterium-mediated transformation of arabidopsis allows efficient plant transformation without need for tissue culture. to facilitate use with other plant species, we investigated the mechanisms that underlie this method. in manual outcrossing experiments, application of agrobacterium tumefaciens to pollen donor plants did not produce any transformed progeny, whereas application of agrobacterium to pollen recipient plants yielded transformants at a rate of 0.48%. ag ...200010889238
a bacterial sensor of plant cell contact controls the transcriptional induction of ralstonia solanacearum pathogenicity genes.the hrp genes of the plant pathogen ralstonia solanacearum are key pathogenicity determinants; they encode a type iii protein secretion machinery involved in the secretion of mediators of the bacterium-plant interaction. these hrp genes are under the genetic control of the hrpb regulatory gene, expression of which is induced when bacteria are co-cultivated with plant cell suspensions. in this study, we used hrp-gfp transcriptional fusions to demonstrate that the expression of the hrpb and type i ...200010811621
genes expressed in pseudomonas putida during colonization of a plant-pathogenic fungus.in vivo expression technology (ivet) was employed to study colonization of phytophthora parasitica by a biological control bacterium, pseudomonas putida 06909, based on a new selection marker. the pyrb gene, which encodes aspartate transcarbamoylase, an enzyme used for pyrimidine biosynthesis, was cloned from p. putida 06909. a pyrb-disrupted mutant did not grow in pyrimidine-deficient media unless it was complemented with pyrbc' behind an active promoter. thirty clones obtained from p. putida 0 ...200010877766
cultivar-specific avirulence and virulence functions assigned to avrpphf in pseudomonas syringae pv. phaseolicola, the cause of bean halo-blight disease.the avrpphf gene was cloned from pseudomonas syringae pathovar phaseolicola (pph:) races 5 and 7, based on its ability to confer avirulence towards bean cultivars carrying the r1 gene for halo-blight resistance, such as red mexican. avrpphf comprised two open reading frames, which were both required for function, and was located on a 154 kb plasmid (pav511) in pph: strain rw60 of pph:, lacking pav511, displayed a loss in virulence to a range of previously susceptible cultivars such as tendergree ...200010880434
arabidopsis thaliana eds4 contributes to salicylic acid (sa)-dependent expression of defense responses: evidence for inhibition of jasmonic acid signaling by sa.the arabidopsis enhanced disease susceptibility 4 (eds4) mutation causes enhanced susceptibility to infection by the bacterial pathogen pseudomonas syringae pv. maculicola es4326 (psm es4326). gene-for-gene resistance to bacteria carrying the avirulence gene avrrpt2 is not significantly affected by eds4. plants homozygous for eds4 exhibit reduced expression of the pathogenesis-related gene pr-1 after infection by psm es4326, weakened responses to treatment with the signal molecule salicylic acid ...200010796016
eukaryotic fatty acylation drives plasma membrane targeting and enhances function of several type iii effector proteins from pseudomonas syringae.bacterial pathogens of plants and animals utilize conserved type iii delivery systems to traffic effector proteins into host cells. plant innate immune systems evolved disease resistance (r) genes to recognize some type iii effectors, termed avirulence (avr) proteins. on disease-susceptible (r) plants, avr proteins can contribute to pathogen virulence. we demonstrate that several type iii effectors from pseudomonas syringae are targeted to the host plasma membrane and that efficient membrane ass ...200010830163
prey range characterization, ribotyping, and diversity of soil and rhizosphere bdellovibrio spp. isolated on phytopathogenic bacteria.thirty new bdellovibrio strains were isolated from an agricultural soil and from the rhizosphere of plants grown in that soil. using a combined molecular and culture-based approach, we found that the soil bdellovibrios included subpopulations of organisms that differed from rhizosphere bdellovibrios. thirteen soil and seven common bean rhizosphere bdellovibrio strains were isolated when pseudomonas corrugata was used as prey; seven and two soil strains were isolated when erwinia carotovora subsp ...200010831412
surface-displayed viral antigens on salmonella carrier vaccine.we have developed a recombinant live oral vaccine using the ice-nucleation protein (inp) from pseudomonas syringae to display viral antigens on the surface of salmonella spp. fusion proteins containing viral antigens were expressed in the oral vaccine strain, salmonella typhi ty21a. surface localization was verified by immunoblotting and fluorescence-activated cell sorting. the immunogenicity of surface-displayed viral antigens on the recombinant live vaccine strain was assessed in mice inoculat ...200010835603
ethylene formation and phenotypic analysis of transgenic tobacco plants expressing a bacterial ethylene-forming enzyme.a bacterial ethylene-forming enzyme (efe) catalyzes oxygenation of 2-oxoglutarate to produce ethylene and carbon dioxide in contrast to a plant enzyme which uses 1-aminocyclopropane-1-carboxylic acid as a substrate. we constructed several lines of transgenic tobacco plants which expressed an efe from pseudomonas syringae pv. phaseolicola pk2. the gene encoding a chimeric protein consisting of efe and beta-glucuronidase (gus) was introduced into the tobacco genome using a binary vector which dire ...200010805596
response to xanthomonas campestris pv. vesicatoria in tomato involves regulation of ethylene receptor gene expression.although ethylene regulates a wide range of defense-related genes, its role in plant defense varies greatly among different plant-microbe interactions. we compared ethylene's role in plant response to virulent and avirulent strains of xanthomonas campestris pv. vesicatoria in tomato (lycopersicon esculentum mill.). the ethylene-insensitive never ripe (nr) mutant displays increased tolerance to the virulent strain, while maintaining resistance to the avirulent strain. expression of the ethylene r ...200010806227
isolation of the gene for eilp, an elicitor-inducible lrr receptor-like protein, from tobacco by differential display.we screened tobacco genes, which are differentially expressed in response to a fungal elicitor, and have isolated a gene which codes for a leucine-rich repeat (lrr) protein closely related to cf genes in tomato. the eilp (elicitor inducible lrr protein) gene encodes 95 kda protein, which consists of a putative membrane spanning region, 28 leucine-rich repeats and some n-linked glycosylation sites, and shows high homology to cf-2/cf-5 family genes. southern blot analysis revealed the presence of ...200010845459
molecular cloning of a soybean class iii beta-1,3-glucanase gene that is regulated both developmentally and in response to pathogen infection.we isolated and characterized a soybean gene (sgn1) encoding a basic beta-1,3-glucanase that is a plant class iii isoform of beta-1,3-glucanase. the deduced amino acid sequence of the sgn1 gene is similar to that of the pr-q'b gene, the basic class iii beta-1,3-glucanase of tomato. based on rna blot hybridization, sgn1 gene expression was detected in all tissues of 4-day old seedlings, but it was present only in root tissue of 30-day old plants. gus expression analysis carried out in transgenic ...200010725560
members of the arabidopsis hrt/rpp8 family of resistance genes confer resistance to both viral and oomycete pathogens.turnip crinkle virus (tcv) inoculation onto tcv-resistant arabidopsis leads to a hypersensitive response (hr) controlled by the dominant gene hrt. hrt is a member of the class of resistance (r) genes that contain a leucine zipper, a nucleotide binding site, and leucine-rich repeats. the chromosomal position of hrt and its homology to resistance gene rpp8 and two rpp8 homologs indicate that unequal crossing over and gene conversion may have contributed to hrt evolution. rpp8 confers resistance to ...200010810142
pti4 is induced by ethylene and salicylic acid, and its product is phosphorylated by the pto kinase.the tomato pti4 gene encodes a transcription factor that was identified on the basis of its specific interaction with the product of the pto disease resistance gene in a yeast two-hybrid system. we show here that the pti4 protein specifically binds the gcc-box cis element, which is present in the promoter region of many pathogenesis-related (pr) genes. expression of the pti4 gene in tomato leaves was rapidly induced by ethylene and by infection with pseudomonas syringae pv tomato, and this induc ...200010810149
a resistance gene product of the nucleotide binding site -- leucine rich repeats class can form a complex with bacterial avirulence proteins in vivo.resistance (r) genes in plants mediate gene-for-gene disease resistance. the ligand-receptor model, which explains the gene-for-gene specificity, predicts a physical interaction between an elicitor, which is directly or indirectly encoded by an avirulence (avr) gene in the pathogen, and the corresponding r gene product. the nucleotide binding site (nbs) - leucine rich repeats (lrr) class of r genes is the largest known class of r genes. here we report that an nbs-lrr r protein and its cognate av ...200010849351
characterization of the fructosyltransferase gene of actinomyces naeslundii wvu45.oral actinomycetes produce fructosyltransferase (ftf) enzymes which convert sucrose into polymers of d-fructose, known as levans, and these polymers are thought to contribute to the persistence and virulence of the organisms. a gene encoding ftf was isolated from actinomyces naeslundii wvu45; the deduced amino acid sequence showed significant similarity to known levansucrases of gram-negative environmental isolates but was less similar to ftfs from gram-positive bacteria. a transcriptional start ...200010850978
sequence analysis and initial characterization of two isozymes of hydroxylaminobenzene mutase from pseudomonas pseudoalcaligenes js45.pseudomonas pseudoalcaligenes js45 grows on nitrobenzene by a partially reductive pathway in which the intermediate hydroxylaminobenzene is enzymatically rearranged to 2-aminophenol by hydroxylaminobenzene mutase (hab mutase). the properties of the enzyme, the reaction mechanism, and the evolutionary origin of the gene(s) encoding the enzyme are unknown. in this study, two open reading frames (haba and habb), each encoding an hab mutase enzyme, were cloned from a p. pseudoalcaligenes js45 genomi ...200010877793
nitric oxide and salicylic acid signaling in plant defense.salicylic acid (sa) plays a critical signaling role in the activation of plant defense responses after pathogen attack. we have identified several potential components of the sa signaling pathway, including (i) the h(2)o(2)-scavenging enzymes catalase and ascorbate peroxidase, (ii) a high affinity sa-binding protein (sabp2), (iii) a sa-inducible protein kinase (sipk), (iv) npr1, an ankyrin repeat-containing protein that exhibits limited homology to ikappabalpha and is required for sa signaling, ...200010922045
marinomonas mediterranea mmb-1 transposon mutagenesis: isolation of a multipotent polyphenol oxidase mutant.marinomonas mediterranea is a melanogenic marine bacterium expressing a multifunctional polyphenol oxidase (ppo) able to oxidize substrates characteristic for laccases and tyrosinases, as well as produce a classical tyrosinase. a new and quick method has been developed for screening laccase activity in culture plates to detect mutants differentially affected in this ppo activity. transposon mutagenesis has been applied for the first time to m. mediterranea by using different minitransposons load ...200010850991
virulence of the phytopathogen pseudomonas syringae pv. maculicola is rpon dependent.we cloned the rpon (ntra and glnf) gene encoding sigma(54) from the phytopathogen pseudomonas syringae pv. maculicola strain es4326. the p. syringae es4326 rpon gene complemented pseudomonas aeruginosa, escherichia coli, and klebsiella aerogenes rpon mutants for a variety of rpon mutant phenotypes, including the inability to utilize nitrate as sole nitrogen source. dna sequence analysis of the p. syringae es4326 rpon gene revealed that the deduced amino acid sequence was most similar (86% identi ...200010852883
the alternative sigma factor rpon is required for hrp activity in pseudomonas syringae pv. maculicola and acts at the level of hrpl transcription.beta-glucuronidase (uida) reporter gene fusions were constructed for the hrpz, hrpl, and hrps genes from the phytopathogen pseudomonas syringae pv. maculicola strain es4326. these reporters, as well as an avrrpt2-uida fusion, were used to measure transcriptional activity in es4326 and a es4326 rpon mutant. rpon was required for the expression of avrrpt2, hrpz, and hrpl in vitro in minimal media and in vivo when infiltrated into arabidopsis thaliana leaves. in contrast, the expression of hrps was ...200010852884
cdna-aflp reveals a striking overlap in race-specific resistance and wound response gene expression profiles.the tomato cf-9 gene confers resistance to races of the fungal pathogen cladosporium fulvum expressing the avr9 gene. cdna amplified fragment length polymorphism analysis was used to display transcripts whose expression is rapidly altered during the avr9- and cf-9-mediated defense response in tobacco cell cultures. diphenyleneiodonium was used to abolish the production of active oxygen species during gene induction. of 30,000 fragments inspected, 290 showed altered abundance, of which 263 were i ...200010852940
induced expression of sarcotoxin ia enhanced host resistance against both bacterial and fungal pathogens in transgenic tobacco.we demonstrate here that induced expression of sarcotoxin ia, a bactericidal peptide from sarcophaga peregrina, enhanced the resistance of transgenic tobacco plants to both bacterial and fungal pathogens. the peptide was produced with a modified pr1a promoter, which is further activated by salicylic acid treatment and necrotic lesion formation by pathogen infection. host resistance to infection of bacteria erwinia carotovora subsp. carotovora and pseudomonas syringae pv. tabaci was shown to be d ...200010939257
evidence for a role of the n terminus and leucine-rich repeat region of the mi gene product in regulation of localized cell death.the tomato mi gene confers resistance against root-knot nematodes and potato aphids. chimeric constructs of the functional gene, mi-1. 2, with a homolog, mi-1.1, were produced, and their phenotypes were examined in agrobacterium rhizogenes-transformed roots. exchange of the leucine-rich repeat (lrr) region of mi-1.1 into mi-1.2 resulted in the loss of ability to confer nematode resistance, as did substitution of a 6-amino acid sequence from the mi-1.1 lrr into mi-1.2. introduction of the mi-1.2 ...200010948252
regulation of brucella abortus catalase.all aerobic organisms have mechanisms that protect against oxidative compounds. catalase, peroxidase, superoxide dismutase, glutathione, and thioredoxin are widely distributed in many taxa and constitute elements of a nearly ubiquitous antioxidant metabolic strategy. interestingly, the regulatory mechanisms that control these elements are rather different depending on the nature of the oxidative stress and the organism. catalase is well documented to play an important role in protecting cells fr ...200010858195
a regulatory rna (prrb rna) modulates expression of secondary metabolite genes in pseudomonas fluorescens f113.the gacs-gaca two-component signal transduction system, which is highly conserved in gram-negative bacteria, is required for the production of exoenzymes and secondary metabolites in pseudomonas spp. screening of a pseudomonas fluorescens f113 gene bank led to the isolation of a previously undefined locus which could restore secondary metabolite production to both gacs and gaca mutants of f113. sequence analysis of this locus demonstrated that it did not contain any obvious pseudomonas protein-c ...200010869066
chemoenzymatic synthesis of the microbial elicitor (-)-syringolide via a fructose 1,6-diphosphate aldolase-catalyzed condensation reaction.syringolide 2, an elicitor of the bacterial plant pathogen pseudomonas syringae pv. tomato which triggers a hypersensitive defense response in resistant soybean plants, has been synthesized in five steps via a fructose 1,6-diphosphate aldolase reaction.200010959854
a new two-component regulatory system involved in adhesion, autolysis, and extracellular proteolytic activity of staphylococcus aureus.a transposition mutant of staphylococcus aureus was selected from the parent strain mt23142, a derivative of strain 8325. the site of transposition was near the 5' terminus of the gene arls. arls exhibits strong similarities with histidine protein kinases. sequence analysis suggested that arls forms an operon with upstream gene arlr. the predicted product of arlr is a member of the ompr-phob family of response regulators. the arls mutant formed a biofilm on a polystyrene surface unlike the paren ...200010869073
applications of differential-display reverse transcription-pcr to molecular pathogenesis and medical mycology.the host-fungus interaction is characterized by changes in gene expression in both host and pathogen. differential-display reverse transcription pcr (ddrt-pcr) is a pcr-based method that allows extensive analysis of gene expression among several cell populations. several limitations and drawbacks to this procedure have now been addressed, including the large number of false-positive results and the difficulty in confirming differential expression. modifications that simplify the reaction time, a ...200010885984
enhancement of induced disease resistance by simultaneous activation of salicylate- and jasmonate-dependent defense pathways in arabidopsis thaliana.the plant-signaling molecules salicylic acid (sa) and jasmonic acid (ja) play an important role in induced disease resistance pathways. cross-talk between sa- and ja-dependent pathways can result in inhibition of ja-mediated defense responses. we investigated possible antagonistic interactions between the sa-dependent systemic acquired resistance (sar) pathway, which is induced upon pathogen infection, and the ja-dependent induced systemic resistance (isr) pathway, which is triggered by nonpatho ...200010890883
the octadecanoid pathway is required for pathogen-induced multi-functional acetyl-coa carboxylase accumulation in common bean (phaseolus vulgaris l.).a partial cdna clone corresponding to the multi-functional acetyl-coa carboxylase (accase, ec 6.4.1.2) was isolated using rna extracted from methyl jasmonate (meja)-induced common bean cell cultures. most of this clone corresponds to the 3' untranslated region and it showed high identity to alfalfa and soybean accase sequences. southern hybridization revealed one copy of this gene in the common bean genome. in addition to being induced by meja in cell cultures and leaves, accase mrna accumulated ...200010960731
genotypic characterization of bradyrhizobium strains nodulating small senegalese legumes by 16s-23s rrna intergenic gene spacers and amplified fragment length polymorphism fingerprint analyses.we examined the genotypic diversity of 64 bradyrhizobium strains isolated from nodules from 27 native leguminous plant species in senegal (west africa) belonging to the genera abrus, alysicarpus, bryaspis, chamaecrista, cassia, crotalaria, desmodium, eriosema, indigofera, moghania, rhynchosia, sesbania, tephrosia, and zornia, which play an ecological role and have agronomic potential in arid regions. the strains were characterized by intergenic spacer (between 16s and 23s rrna genes) pcr and res ...200010966419
new substrates for the dicarboxylate transport system of sinorhizobium meliloti.the dicarboxylate transport (dct) system of sinorhizobium meliloti, which is essential for a functional nitrogen-fixing symbiosis, has been thought to transport only dicarboxylic acids. we show here that the permease component of the dct system, dcta, can transport orotate, a monocarboxylic acid, with an apparent k(m) of 1.7 mm and a v(max) of 163 nmol min(-1) per mg of protein in induced cells. dcta was not induced by the presence of orotate. the transport of orotate was inhibited by several co ...200010894729
ice crystallization by pseudomonas syringae.several bacterial species can serve as biological ice nuclei. the best characterized of these is pseudomonas syringae, a widely distributed bacterial epiphyte of plants. these biological ice nuclei find various applications in different fields, but an optimized production method was required in order to obtain the highly active cells which may be exploited as ice nucleators. the results presented here show that p. syringae cells reduce supercooling of liquid or solid media and enhance ice crysta ...200010968626
a unique 33-kd cysteine proteinase accumulates in response to larval feeding in maize genotypes resistant to fall armyworm and other lepidoptera.plants respond to insect feeding with a number of defense mechanisms. using maize genotypes derived from antiquan germ plasm that are resistant to lepidoptera, we have demonstrated that a unique 33-kd cysteine proteinase accumulates in the whorl in response to larval feeding. the abundance of the proteinase increased dramatically at the site of larval feeding after 1 hr of infestation and continued to accumulate for as long as 7 days. the 33-kd cysteine proteinase was most abundant in the yellow ...200010899972
the hypersensitive response facilitates plant infection by the necrotrophic pathogen botrytis cinerea.plants have evolved efficient mechanisms to combat pathogen attack. one of the earliest responses to attempted pathogen attack is the generation of oxidative burst that can trigger hypersensitive cell death. this is called the hypersensitive response (hr) and is considered to be a major element of plant disease resistance. the hr is thought to deprive the pathogens of a supply of food and confine them to initial infection site. necrotrophic pathogens, such as the fungi botrytis cinerea and scler ...200010898976
the arabidopsis dnd1 "defense, no death" gene encodes a mutated cyclic nucleotide-gated ion channel.gene-for-gene disease resistance typically includes a programmed cell death response known as the hypersensitive response (hr). the arabidopsis thaliana dnd1 mutant was previously isolated as a line that failed to produce the hr in response to avirulent pseudomonas syringae pathogens; plants homozygous for the recessive dnd1-1 mutation still carry out effective gene-for-gene resistance. the dnd1-1 mutation also causes constitutive systemic resistance and elevated levels of salicylic acid. in the ...200010900264
bacteria in the leaf ecosystem with emphasis on pseudomonas syringae-a pathogen, ice nucleus, and epiphyte.the extremely large number of leaves produced by terrestrial and aquatic plants provide habitats for colonization by a diversity of microorganisms. this review focuses on the bacterial component of leaf microbial communities, with emphasis on pseudomonas syringae-a species that participates in leaf ecosystems as a pathogen, ice nucleus, and epiphyte. among the diversity of bacteria that colonize leaves, none has received wider attention than p. syringae, as it gained notoriety for being the firs ...200010974129
ankb, a periplasmic ankyrin-like protein in pseudomonas aeruginosa, is required for optimal catalase b (katb) activity and resistance to hydrogen peroxide.in this study, we have cloned the ankb gene, encoding an ankyrin-like protein in pseudomonas aeruginosa. the ankb gene is composed of 549 bp encoding a protein of 183 amino acids that possesses four 33-amino-acid ankyrin repeats that are a hallmark of erythrocyte and brain ankyrins. the location of ankb is 57 bp downstream of katb, encoding a hydrogen peroxide-inducible catalase, katb. monomeric ankb is a 19.4-kda protein with a pi of 5.5 that possesses 22 primarily hydrophobic amino acids at re ...200010913088
controlling instability in gacs-gaca regulatory genes during inoculant production of pseudomonas fluorescens biocontrol strains.secondary metabolism in fluorescent pseudomonads is globally regulated by gacs, which encodes a membrane-bound sensor kinase, and gaca, which encodes a transcriptional response regulator. spontaneous mutation in either gene blocked biosynthesis of the antimicrobial compounds hydrogen cyanide, 2,4-diacetylphloroglucinol, pyoluteorin, and pyrrolnitrin by the model biocontrol strain pseudomonas fluorescens cha0. spontaneous mutants also had altered abilities to utilize several carbon sources and to ...200010919762
a genomic sample sequence of the entomopathogenic bacterium photorhabdus luminescens w14: potential implications for virulence.photorhabdus luminescens is a pathogenic bacterium that lives in the guts of insect-pathogenic nematodes. after invasion of an insect host by a nematode, bacteria are released from the nematode gut and help kill the insect, in which both the bacteria and the nematodes subsequently replicate. however, the bacterial virulence factors associated with this "symbiosis of pathogens" remain largely obscure. in order to identify genes encoding potential virulence factors, we performed approximately 2,00 ...200010919786
in vitro antimicrobial properties of recombinant asabf, an antimicrobial peptide isolated from the nematode ascaris suum.asabf is a csalphabeta-type antimicrobial peptide that contains four intramolecular disulfide bridges (y. kato and s. komatsu, j. biol. chem. 271:30493-30498, 1996). in the present study, a recombinant asabf was produced by using a yeast expression system, and its antimicrobial activity was characterized in detail. the recombinant asabf was active against all gram-positive bacteria tested (7 of 7; minimum bactericidal concentration [mbc], 0.03 to 1 microg/ml) except leuconostoc mesenteroides, so ...200010991847
rapid methods for testing the efficacy of sterilization-grade filter membranes.the validation of sterilization-grade membranes is integral to ensuring the efficient and safe use of microfiltration systems. here validation refers to the production of sterile filtrate for sterilizing-grade membranes under challenge test conditions. current validation methods require 48 h of culture for results to become available, which creates time delays within the manufacturing process and quality control (qc) backlogs. this work compares four methods for the production of filter challeng ...200010919803
expression of a functional antizearalenone single-chain fv antibody in transgenic arabidopsis plants.the efficacy of cloning a recombinant mycotoxin antibody in plants was tested using arabidopsis as a model. an antizearalenone single-chain fv (scfv) dna fragment was first cloned in the newly constructed phage display vector (pey.5) and then recloned in the plant transformation vector pkylx71::35s(2). after transformation, constructs of antizearalenone scfv were introduced into immature arabidopsis seeds via agrobacterium tumefaciens mediation by vacuum infiltration. only plants transformed wit ...200010919813
pseudomonas syringae hrp type iii secretion system and effector proteins.pseudomonas syringae is a member of an important group of gram-negative bacterial pathogens of plants and animals that depend on a type iii secretion system to inject virulence effector proteins into host cells. in p. syringae, hrp/hrc genes encode the hrp (type iii secretion) system, and avirulence (avr) and hrp-dependent outer protein (hop) genes encode effector proteins. the hrp/hrc genes of p. syringae pv syringae 61, p. syringae pv syringae b728a, and p. syringae pv tomato dc3000 are flanke ...200010922033
genetic manipulation of isoflavone 7-o-methyltransferase enhances biosynthesis of 4'-o-methylated isoflavonoid phytoalexins and disease resistance in alfalfa.4'-o-methylation of an isoflavonoid intermediate is a key reaction in the biosynthesis of the phytoalexin medicarpin in legumes. however, isoflavone o-methyltransferase (iomt) from alfalfa converts the isoflavone daidzein to 7-o-methyl daidzein (isoformononetin) in vitro as well as in vivo in unchallenged leaves of transgenic alfalfa ectopically expressing iomt. in contrast, elicitation of iomt-overexpressing plants with cucl(2) or infecting these plants with phoma medicaginis leads to greater a ...200011006341
molecular and cell biology aspects of plague.a 70-kb virulence plasmid (sometimes called pyv) enables yersinia spp. to survive and multiply in the lymphoid tissues of their host. it encodes the yop virulon, a system consisting of secreted proteins called yops and their dedicated type iii secretion apparatus called ysc. the ysc apparatus forms a channel composed of 29 proteins. of these, 10 have counterparts in almost every type iii system. secretion of some yops requires the assistance, in the bacterial cytosol, of small individual chapero ...200010922034
avrpto-dependent pto-interacting proteins and avrpto-interacting proteins in tomato.the plant-intracellular interaction of the avirulence protein avrpto of pseudomonas syringae pathovar tomato, the agent of bacterial speck disease, and the corresponding tomato resistance protein pto triggers responses leading to disease resistance. pto, a serine/threonine protein kinase, also interacts with a putative downstream kinase, pto-interactor 1, as well as with members of a family of transcription factors pto-interactors 4, 5, and 6. these proteins are likely involved, respectively, in ...200010922043
pcr assays that identify the grapevine dieback fungus eutypa lata.eutypa lata is the causal fungal agent of eutypa dieback, a serious grapevine necrotic disease. the erratic and delayed (1 to 2 months) appearance of characteristic conidia on culture media and the presence of numerous microorganisms in decaying wood make it difficult either to identify or to detect e. lata in grapevine wood samples. we designed six pairs of pcr primers for diagnosis of e. lata. three primer pairs were derived from ribosomal dna internal transcribed spacer sequences, and three p ...200011010901
requirement of sphingolipid alpha-hydroxylation for fungicidal action of syringomycin e.syringomycin e is an antifungal cyclic lipodepsinonapeptide produced by pseudomonas syringae pv. syringae. to understand the mechanism of action of syringomycin e, a novel resistant saccharomyces cerevisiae strain, bw7, was isolated and characterized. lipid analyses revealed that bw7 contained only the hydrophobic subspecies of sphingolipids that are normally minor components in wild type strains. this aberrant sphingolipid composition was the result of lack of alpha-hydroxylation of the amide-l ...200010922463
conversion of compatible plant-pathogen interactions into incompatible interactions by expression of the pseudomonas syringae pv. syringae 61 hrma gene in transgenic tobacco plants.the hrma gene from pseudomonas syringae pv. syringae has previously been shown to confer avirulence on the virulent bacterium p. syringae pv. tabaci in all examined tobacco cultivars. we expressed this gene in tobacco plants under the control of the tobacco delta0. 3 tobrb7 promoter, which is induced upon nematode infection in tobacco roots (opperman et al. 1994, science, 263, 221-223). a basal level of hrma expression in leaves of transgenic plants activated the expression of pathogenesis-relat ...200010929114
effect of starvation and the viable-but-nonculturable state on green fluorescent protein (gfp) fluorescence in gfp-tagged pseudomonas fluorescens a506.the green fluorescent protein (gfp) gene, gfp, of the jellyfish aequorea victoria is being used as a reporter system for gene expression and as a marker for tracking prokaryotes and eukaryotes. cells that have been genetically altered with the gfp gene produce a protein that fluoresces when it is excited by uv light. this unique phenotype allows gfp-tagged cells to be specifically monitored by nondestructive means. in this study we determined whether a gfp-tagged strain of pseudomonas fluorescen ...200010919764
direct interaction of resistance gene and avirulence gene products confers rice blast resistance.rice expressing the pi-ta gene is resistant to strains of the rice blast fungus, magnaporthe grisea, expressing avr-pita in a gene-for-gene relationship. pi-ta encodes a putative cytoplasmic receptor with a centrally localized nucleotide-binding site and leucine-rich domain (lrd) at the c-terminus. avr-pita is predicted to encode a metalloprotease with an n-terminal secretory signal and pro-protein sequences. avr-pita(176) lacks the secretory and pro-protein sequences. we report here that transi ...200010921881
plants and animals share functionally common bacterial virulence factors.by exploiting the ability of pseudomonas aeruginosa to infect a variety of vertebrate and nonvertebrate hosts, we have developed model systems that use plants and nematodes as adjuncts to mammalian models to help elucidate the molecular basis of p. aeruginosa pathogenesis. our studies reveal a remarkable degree of conservation in the virulence mechanisms used by p. aeruginosa to infect hosts of divergent evolutionary origins.200010922040
the virulence factor avrxa7 of xanthomonas oryzae pv. oryzae is a type iii secretion pathway-dependent nuclear-localized double-stranded dna-binding protein.avrxa7 is a member of the avrbs3 avirulence gene family, which encodes proteins targeted to plant cells by a type iii secretion apparatus. avrxa7, the product of avrxa7, is also a virulence factor in strain pxo86 of xanthomonas oryzae pv. oryzae. avirulence and virulence specificities are associated with the central repeat domain, which, in avrxa7, consists of 25.5 direct repeat units. mutations in three c-terminal nuclear localization signal motifs eliminated avirulence and virulence activities ...200010931960
expression of single chain antibodies (scfvs) for c-myc oncoprotein in recombinant escherichia coli membranes by using the ice-nucleation protein of pseudomonas syringae.the ice nucleation protein (inp) is a glycosyl phosphatidylinositol anchored outer membrane protein found in certain gram-negative bacteria. in this study, the inp from pseudomonas syringae was applied as a fusion partner with the single chain antibody fragment (scfv) against the human oncoprotein c-myc. two new plasmids pninaz-myc and pninazscfv-bsaa1 were constructed and cloned into escherichia coli jm109. the expression of the fusion protein was successfully demonstrated in the cloned cells. ...200010933828
regulation of the rulab mutagenic dna repair operon of pseudomonas syringae by uv-b (290 to 320 nanometers) radiation and analysis of rulab-mediated mutability in vitro and in planta.the effects of the rulab operon of pseudomonas syringae on mutagenic dna repair and the transcriptional regulation of rulab following irradiation with uv-b wavelengths were determined. for a rulb::km insertional mutant constructed in p. syringae pv. syringae b86-17, sensitivity to uv-b irradiation increased and uv mutability decreased by 12- to 14-fold. rulab-induced uv mutability was also tracked in phyllosphere populations of b86-17 for up to 5 days following plant inoculation. uv mutability t ...200011029435
pseudobactin biogenesis in the plant growth-promoting rhizobacterium pseudomonas strain b10: identification and functional analysis of the l-ornithine n(5)-oxygenase (psba) gene.pseudobactin(b10), the fluorescent siderophore produced by the rhizobacterium pseudomonas strain b10, contains the hydroxamate ligand d-n(5)-hydroxyornithine (d-n(5)-oh-orn). we cloned the l-orn n(5)-oxygenase (psba) gene from a genomic library of pseudomonas strain b10 and demonstrated that psba is involved in the conversion of l-orn to its n(5)-oh derivative. psba shows significant similarity to microbial omega-amino acid hydroxylases containing flavin adenine dinucleotide and nadp cofactor-bi ...200011029447
characterization of alginate lyase from pseudomonas syringae pv. syringae.the gene encoding alginate lyase (algl) in pseudomonas syringae pv. syringae was cloned, sequenced, and overexpressed in escherichia coli. alginate lyase activity was optimal when the ph was 7.0 and when assays were conducted at 42 degrees c in the presence of 0.2 m nacl. in substrate specificity studies, algl from p. syringae showed a preference for deacetylated polymannuronic acid. sequence alignment with other alginate lyases revealed conserved regions within algl likely to be important for t ...200011029455
receptor-mediated increase in cytoplasmic free calcium required for activation of pathogen defense in parsley.transient influx of ca(2+) constitutes an early element of signaling cascades triggering pathogen defense responses in plant cells. treatment with the phytophthora sojae-derived oligopeptide elicitor, pep-13, of parsley cells stably expressing apoaequorin revealed a rapid increase in cytoplasmic free calcium ([ca(2+)](cyt)), which peaked at approximately 1 microm and subsequently declined to sustained values of 300 nm. activation of this biphasic [ca(2+)](cyt) signature was achieved by elicitor ...200010948260
cloning and characterization of smedef, a novel multidrug efflux pump from stenotrophomonas maltophilia.stenotrophomonas maltophilia is a nosocomial bacterial pathogen intrinsically resistant to several antibiotics. the mechanisms involved in this intrinsic multiresistance phenotype are poorly understood. a library of chromosomal dna from a spontaneous multidrug-resistant s. maltophilia d457r mutant (a. alonso and j. l. martinez, antimicrob. agents chemother. 41:1140-1142, 1997) was screened for complementation of erythromycin susceptibility on an antibiotic-hypersusceptible escherichia coli delta ...200011036026
constitutive expression of the defense-related rir1b gene in transgenic rice plants confers enhanced resistance to the rice blast fungus magnaporthe grisea.the rirlb gene of rice (oryza sativa) is one of a set of putative defense genes whose transcripts accumulate upon inoculation of rice with the non-host pathogen pseudomonas syringae pv. syringae. it belongs to a family of genes encoding small extracellular proteins so far only identified in cereals. to assess the function of the rirlb gene in rice blast resistance, it was placed under the control of the camv 35s promoter and transferred into rice plants of the japonica variety taipei 309 by biol ...200010949374
mutations in multidrug efflux homologs, sugar isomerases, and antimicrobial biosynthesis genes differentially elevate activity of the sigma(x) and sigma(w) factors in bacillus subtilis.the sigma(x) and sigma(w) extracytoplasmic function sigma factors regulate more than 40 genes in bacillus subtilis. sigma(w) activates genes which function in detoxification and the production of antimicrobial compounds, while sigma(x) activates functions that modify the cell envelope. transposon mutagenesis was used to identify loci which negatively regulate sigma(w) or sigma(x) as judged by up-regulation from the autoregulatory promoter site p(w) or p(x). fourteen insertions that activate p(w) ...200010960106
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