WorldWideScience

Sample records for bacterium erwinia chrysanthemi

  1. Asparaginase Erwinia chrysanthemi

    Science.gov (United States)

    Asparaginase Erwinia chrysanthemi is used with other chemotherapy medications to treat acute lymphocytic leukemia (ALL; a type of cancer ... of allergic reactions to medications similar to asparaginase Erwinia chrysanthemi such as (asparaginase [Elspar] or pegaspargase [Oncaspar]). ...

  2. Ziek en Zeer : Erwinia chrysanthemi in Amaryllidaceae

    NARCIS (Netherlands)

    Vink, P.

    2011-01-01

    In dit artikel een verslag van het onderzoek naar de gevoeligheid van narcissen voor de bacterie Erwinia chrysanthemi (tegenwoordig Dickeya dadantii). Uit een infectieproef is gebleken dat deze bacterie tijdens een reguliere bollenteelt in Lisse niet in staat was om narcisbollen aan te tasten. Toch

  3. Erwinia chrysanthemi: pectolytic bacterium causing soft rot outbreaks of arracacha in Brazil Erwinia chrysanthemi: bactéria pectolítica envolvida na "mela" da mandioquinha-salsa no Brasil

    Directory of Open Access Journals (Sweden)

    Gilmar Paulo Henz

    2006-10-01

    Full Text Available The objetive of this work was to identify the pectolytic bacteria associated with soft rot of arracacha roots in Brazil. From 1998 to 2001, 227 isolates of Erwinia spp. were obtained from arracacha roots and identified by biochemical and physiological tests (pectolytic activity, lecithinase, a-methyl glucoside, phosphatase, erythromycin sensivity, growth at 37ºC. Of these isolates, 89.9% were identified as E. chrysanthemi (Ech, 9.7% as E. carotovora subsp. carotovora (Ecc and 0.5% as E. carotovora subsp. atroseptica. The identity of seventeen out of twenty representative isolates of Ech and Ecc was confirmed by PCR (primers '149f', 'L1r', 'ADE1', 'ADE2'.O objetivo deste trabalho foi identificar as bactérias pectolíticas envolvidas na podridão-mole de raízes de mandioquinha-salsa no Brasil. De 1998 a 2001, 227 isolados de Erwinia spp. foram obtidos de raízes de mandioquinha-salsa e identificados por testes bioquímicos e fisiológicos (atividade pectolítica, lecitinase, a-methyl glucosídeo, fosfatase, sensibilidade à eritromicina, crescimento a 37ºC. Destes isolados, 89,9% foram identificados como E. chrysanthemi (Ech, 9,7% como E. carotovora subsp. carotovora (Ecc e somente 0,5% como E. carotovora subsp. atroseptica. A identidade de 20 isolados representativos de Ech e Ecc foi confirmada por PCR (primers '149f', 'L1r', 'ADE1', 'ADE2', com exceção de dois isolados de Ech e um de Ecc.

  4. Reactive oxygen species activity in the interaction of rice with Erwinia chrysanthemi pv. zeae

    Institute of Scientific and Technical Information of China (English)

    Qiongguang LIU; Landi HE; Jingyi ZHANG; Yutao WANG; Zhenzhong WANG

    2008-01-01

    Activities of reactive oxygen species (ROS) were investigated in the interaction between rice and Erwinia chrysanthemi pv. zeae. Results showed that variety (128) had higher increases in activity compared to those in the susceptible variety (Texian 13) 24 hours after bacteria inoculation. The activities of superoxide dismutase (SOD) increased in 128 and Texian 13 twenty-four hours after inoculation and then decreased, but the SOD activity in 128 was found to be usually lower than that in Texian 13. The CAT activity in Texian 13 had two peaks at 24 h and 96 h after inoculation, while little change was seen in 128. In conclusion, ROS and its related enzymes could be correlated to rice resistance against E. chrysanthemi pv. zeae.

  5. High-throughput screening of Erwinia chrysanthemi pectin methylesterase variants using carbohydrate microarrays

    DEFF Research Database (Denmark)

    Øbro, Jens; Sørensen, Iben; Derkx, Patrick;

    2009-01-01

    Pectin methylesterases (PMEs) catalyse the removal of methyl esters from the homogalacturonan (HG) backbone domain of pectin, a ubiquitous polysaccharide in plant cell walls. The degree of methyl esterification (DE) impacts upon the functional properties of HG within cell walls and plants produce...... numerous PMEs that act upon HG in muro. Many microbial plant pathogens also produce PMEs, the activity of which renders HG more susceptible to cleavage by pectin lyase and polygalacturonase enzymes and hence aids cell wall degradation. We have developed a novel microarray-based approach to investigate...... the activity of a series of variant enzymes based on the PME from the important pathogen Erwinia chrysanthemi. A library of 99 E. chrysanthemi PME mutants was created in which seven amino acids were altered by various different substitutions. Each mutant PME was incubated with a highly methyl esterified lime...

  6. Bacterial leaf rot of Aloe vera L., caused byErwinia chrysanthemi biovar 3

    NARCIS (Netherlands)

    Laat, de P.C.A.; Verhoeven, J.T.W.; Danse, J.D.

    1994-01-01

    A severe attack of the bacteriumErwinia chrysantemi biovar 3 on the succulentAloe vera on the Carribean island of Aruba is described. Biochemical and pathological characteristics of strains are presented, including results of successful inoculation experiments onAloe vera. This is the first report o

  7. 菊欧文氏菌分子检测技术的研究%Molecular detection of Erwinia chrysanthemi

    Institute of Scientific and Technical Information of China (English)

    刘鹏; 黄国明; 刘勇; 崔铁军

    2007-01-01

    蝴蝶兰细菌性软腐病对蝴蝶兰的生长危害严重,Erwinia chrysanthemi(菊欧文氏菌)、Erwinia carotovora subsp.carotovora(胡萝卜软腐欧文氏菌胡萝卜软腐亚种)是引起蝴蝶兰软腐病的主要病原细菌,其中E. chrysanthemi被列入我国三类检疫性有害生物.本文对菊欧文氏菌分子检测技术进行了研究,设计出针对该病原细菌的特异性引物,应用实时荧光PCR方法检测样品中存在的菊欧文氏菌,检测灵敏度达到102 cfu/mL.

  8. The acyl-homoserine lactone-type quorum-sensing system modulates cell motility and virulence of Erwinia chrysanthemi pv. zeae.

    Science.gov (United States)

    Hussain, Mumtaz B B M; Zhang, Hai-Bao; Xu, Jin-Ling; Liu, Qiongguang; Jiang, Zide; Zhang, Lian-Hui

    2008-02-01

    Erwinia chrysanthemi pv. zeae is one of the Erwinia chrysanthemi pathovars that infects on both dicotyledons and monocotyledons. However, little is known about the molecular basis and regulatory mechanisms of its virulence. By using a transposon mutagenesis approach, we cloned the genes coding for an E. chrysanthemi pv. zeae synthase of acyl-homoserine lactone (AHL) quorum-sensing signals (expI(Ecz)) and a cognate response regulator (expR(Ecz)). Chromatography analysis showed that expI(Ecz) encoded production of the AHL signal N-(3-oxo-hexanoyl)-homoserine lactone (OHHL). Null mutation of expI(Ecz) in the E. chrysanthemi pv. zeae strain EC1 abolished AHL production, increased bacterial swimming and swarming motility, disabled formation of multicell aggregates, and attenuated virulence of the pathogen on potato tubers. The mutation also marginally reduced the inhibitory activity of E. chrysanthemi pv. zeae on rice seed germination. The mutant phenotypes were rescued by either exogenous addition of AHL signal or in trans expression of expI(Ecz). These data demonstrate that the AHL-type QS signal plays an essential role in modulation of E. chrysanthemi pv. zeae cell motility and the ability to form multicell aggregates and is involved in regulation of bacterial virulence.

  9. SecG is required for antibiotic activities of Pseudomonas sp. YL23 against Erwinia amylovora and Dickeya chrysanthemi.

    Science.gov (United States)

    Liu, Youzhou; Baird, Sonya M; Qiao, Junqing; Du, Yan; Lu, Shi-En

    2015-05-01

    Strain YL23 was isolated from soybean root tips and identified to be Pseudomonas sp. This strain showed broad-spectrum antibacterial activity against bacterial pathogens that are economically important in agriculture. To characterize the genes dedicated to antibacterial activities against microbial phytopathogens, a Tn5-mutation library of YL23 was constructed. Plate bioassays revealed that the mutant YL23-93 lost its antibacterial activities against Erwinia amylovora and Dickeya chrysanthemi as compared with its wild type strain. Genetic and sequencing analyses localized the transposon in a homolog of the secG gene in the mutant YL23-93. Constitutive expression plasmid pUCP26-secG was constructed and electroporated into the mutant YL23-93. Introduction of the plasmid pUCP26-secG restored antibacterial activities of the mutant YL23-93 to E. amylovora and D. chrysanthemi. As expected, empty plasmid pUCP26 could not complement the phenotype of the antibacterial activity in the mutant. Thus the secG gene, belonging to the Sec protein translocation system, is required for antibacterial activity of strain YL23 against E. amylovora and D. chrysanthemi.

  10. Tol-Pal proteins are critical cell envelope components of Erwinia chrysanthemi affecting cell morphology and virulence.

    Science.gov (United States)

    Dubuisson, Jean-François; Vianney, Anne; Hugouvieux-Cotte-Pattat, Nicole; Lazzaroni, Jean Claude

    2005-10-01

    The tol-pal genes are necessary for maintaining the outer-membrane integrity of Gram-negative bacteria. These genes were first described in Escherichia coli, and more recently in several other species. They are involved in the pathogenesis of E. coli, Haemophilus ducreyi, Vibrio cholerae and Salmonella enterica. The role of the tol-pal genes in bacterial pathogenesis was investigated in the phytopathogenic enterobacterium Erwinia chrysanthemi, assuming that this organism might be a good model for such a study. The whole Er. chrysanthemi tol-pal region was characterized. Tol-Pal proteins, except TolA, showed high identity scores with their E. coli homologues. Er. chrysanthemi mutants were constructed by introducing a uidA-kan cassette in the ybgC, tolQ, tolA, tolB, pal and ybgF genes. All the mutants were hypersensitive to bile salts. Mutations in tolQ, tolA, tolB and pal were deleterious for the bacteria, which required high concentrations of sugars or osmoprotectants for their viability. Consistent with this observation, they were greatly impaired in their cell morphology and division, which was evidenced by observations of cell filaments, spherical forms, membrane blebbing and mislocalized bacterial septa. Moreover, tol-pal mutants showed a reduced virulence in a potato tuber model and on chicory leaves. This could be explained by a combination of impaired phenotypes in the tol-pal mutants, such as reduced growth and motility and a decreased production of pectate lyases, the major virulence factor of Er. chrysanthemi.

  11. Asparaginase Erwinia chrysanthemi (Erwinaze®): a guide to its use in acute lymphoblastic leukemia in the USA.

    Science.gov (United States)

    Keating, Gillian M

    2013-08-01

    Asparaginase Erwinia chrysanthemi (Erwinaze®) is approved in the USA for use in patients with acute lymphoblastic leukemia (ALL) who have developed hypersensitivity to Escherichia coli-derived asparaginase. The approved regimen of intramuscular Erwinaze® was associated with sustained, clinically meaningful asparaginase activity in patients with ALL who had to discontinue treatment with pegaspargase (a pegylated formulation of E. coli asparaginase) because of hypersensitivity. Another study revealed that development of E. coli-derived asparaginase allergy and a switch to Erwinaze® maintained event-free survival in pediatric patients with newly diagnosed ALL. In a multicenter, compassionate-use trial, Erwinaze® was generally well tolerated, with the most commonly occurring adverse events including hypersensitivity, pancreatitis, fever, hyperglycemia, and increased transaminase levels. Subclinical hypersensitivity may result in the inactivation of asparaginase and affect treatment outcome; monitoring of serum asparaginase levels may be used to identify subclinical hypersensitivity.

  12. Some of the out genes involved in the secretion of pectate lyases in Erwinia chrysanthemi are regulated by kdgR.

    Science.gov (United States)

    Condemine, G; Dorel, C; Hugouvieux-Cotte-Pattat, N; Robert-Baudouy, J

    1992-11-01

    The out genes of Erwinia chrysanthemi are required for the translocation across the outer membrane of pectate lyases and cellulases. We present the characterization and the nucleotide sequence of five genes of the out cluster. The products of outS, B, C, D and E have significant homology with the PulS, B, C, D and E proteins necessary to the secretion of pullulanase in Klebsiella pneumoniae. An open reading frame, outT, located between outB and outC has no homology with the pul cluster but is involved in secretion. outC, outD and outE form an operon while outS, outB and outT constitute independent transcription units. outT and the outCDE operon are regulated by kdgR, the negative regulatory gene controlling pectinase production. outB and outS seem to be expressed constitutively.

  13. The genome of Erwinia tasmaniensis strain Et1/99, a non-pathogenic bacterium in the genus Erwinia.

    Science.gov (United States)

    Kube, Michael; Migdoll, Alexander Michael; Müller, Ines; Kuhl, Heiner; Beck, Alfred; Reinhardt, Richard; Geider, Klaus

    2008-09-01

    The complete genome of the bacterium Erwinia tasmaniensis strain Et1/99 consisting of a 3.9 Mb circular chromosome and five plasmids was sequenced. Strain Et1/99 represents an epiphytic plant bacterium related to Erwinia amylovora and E. pyrifoliae, which are responsible for the important plant diseases fire blight and Asian pear shoot blight, respectively. Strain Et1/99 is a non-pathogenic bacterium and is thought to compete with these and other bacteria when occupying the same habitat during initial colonization. Genome analysis revealed tools for colonization, cellular communication and defence modulation, as well as genes coding for the synthesis of levan and a not detected capsular exopolysaccharide. Strain Et1/99 may secrete indole-3-acetic acid to increase availability of nutrients provided on plant surfaces. These nutrients are subsequently accessed and metabolized. Secretion systems include the hypersensitive response type III pathway present in many pathogens. Differences or missing parts within the virulence-related factors distinguish strain Et1/99 from pathogens such as Pectobacterium atrosepticum and the related Erwinia spp. Strain Et1/99 completely lacks the sorbitol operon, which may also affect its inability to invade fire blight host plants. Erwinia amylovora in contrast depends for virulence on utilization of sorbitol, the dominant carbohydrate in rosaceous plants. The presence of other virulence-associated factors in strain Et1/99 indicates the ancestral genomic background of many plant-associated bacteria.

  14. Reinfection and latent infection of Erwinia chrysanthemi pv.zeae in rice%水稻细菌性基腐病菌再侵染和潜伏侵染

    Institute of Scientific and Technical Information of China (English)

    刘琼光; 王振中; 陈玉托; 区伟明; 区肇康

    2003-01-01

    @@ 水稻细菌性基腐病(Erwinia chrysanthemi pv.zeae)近年来在我国局部地区发病加重.有关病菌生物学特性和侵染规律有过一些研究和报道,但病害后期表现症状及病菌的再侵染和潜伏侵染现象等,均未见报道,笔者对此进行了研究.

  15. Erwinia chrysanthemi分离株CSCL006 hrpN基因的克隆与高效表达%Molecular Characterization and Overexpression of Erwinia chrysanthemi Strain CSCL006 hrpNCSCL006 Gene, which Encodes An Elicitor of the Hyperdensitive Reaction

    Institute of Scientific and Technical Information of China (English)

    汤承; 崔亚亚; 吴伯骥; 李名扬

    2006-01-01

    通过构建Erwinia chrysanthemi分离株CSCL006的DNA文库,克隆出hrpN CSCL006基因,测序结果显示该基因编码区长1 020 bp;推导的harpinCSCL006与Erwinia chrysanthemi ECl6和3937编码的harpin蛋白同源性高,但与其它软腐菌的harpin蛋白同源性较低;在大肠杆菌中(Escherichia coli)高效表达了hrpN CSCL006基因,重组harpinCSCL006蛋白分子量为34kD.以抗harpinEOC的抗体为探针,Western blot证实该蛋白确为harpin;纯化的harpinCSCL006,能引起烟叶的过敏反应.

  16. HecA, a member of a class of adhesins produced by diverse pathogenic bacteria, contributes to the attachment, aggregation, epidermal cell killing, and virulence phenotypes of Erwinia chrysanthemi EC16 on Nicotiana clevelandii seedlings.

    Science.gov (United States)

    Rojas, Clemencia M; Ham, Jong Hyun; Deng, Wen-Ling; Doyle, Jeff J; Collmer, Alan

    2002-10-01

    Erwinia chrysanthemi is representative of a broad class of bacterial pathogens that are capable of inducing necrosis in plants. The E. chrysanthemi EC16 hecA gene predicts a 3,850-aa member of the Bordetella pertussis filamentous hemagglutinin family of adhesins. A hecATn7 mutant was reduced in virulence on Nicotiana clevelandii seedlings after inoculation without wounding. Epifluorescence and confocal laser-scanning microscopy observations of hecA and wild-type cells expressing the green fluorescent protein revealed that the mutant is reduced in its ability to attach and then form aggregates on leaves and to cause an aggregate-associated killing of epidermal cells. Cell killing also depended on production of the major pectate lyase isozymes and the type II, but not the type III, secretion pathway in E. chrysanthemi. HecA homologs were found in bacterial pathogens of plants and animals and appear to be unique to pathogens and universal in necrogenic plant pathogens. Phylogenetic comparison of the conserved two-partner secretion domains in the proteins and the 16S rRNA sequences in respective bacteria revealed the two datasets to be fundamentally incongruent, suggesting horizontal acquisition of these genes. Furthermore, hecA and its two homologs in Yersinia pestis had a G+C content that was 10% higher than that of their genomes and similar to that of plant pathogenic Ralstonia, Xylella, and Pseudomonas spp. Our data suggest that filamentous hemagglutinin-like adhesins are broadly important virulence factors in both plant and animal pathogens.

  17. 玉米细菌性茎腐病的发生为害调查%Surveys of the occurrence and damage of corn bacterial stalk rots (Erwinia chrysanthemi pv.zeae)

    Institute of Scientific and Technical Information of China (English)

    李巧芝; 高明; 王自伟; 高清珍; 罗秦岳

    2002-01-01

    系统调查表明,玉米细菌性茎腐病由玉米细菌性茎腐病菌 (Erwinia chrysanthemi pv.zeae)所致,在玉米植株中部叶鞘和茎秆上发生水渍状腐烂,引起组织软化.病菌随病残组织在田间、地边越冬.夏季暴雨多、空气湿度大、虫害发生重等对病害发生有利.提出及时清除病残体、防治虫害、适时施药的防治措施.

  18. An Endophytic Erwinia chrysanthemi Strain Antagonistic against Banana Fusarium Wilt Disease%一株对香蕉枯萎病菌具有良好拮抗作用的菊欧氏杆菌

    Institute of Scientific and Technical Information of China (English)

    殷晓敏; 陈弟; 吴红萍; 郑服丛

    2009-01-01

    在香蕉枯萎病重病园区,从生长正常香蕉假茎内分离获得一株细菌E353菌株.经对峙培养、孢子萌发抑制测定,E353对香蕉枯萎病菌菌丝生长、孢子萌发具有良好抑制效果.盆栽试验表明,E353活菌培养液(750ml/株)浸根处理,对香蕉枯萎病的防效为60.67%.经形态学、生理生化和16S rDNA序列比对,将E353鉴定为菊欧氏杆菌Erwinia chrysanthemi.

  19. Pathogenicity and Biological Characters of Erwinia chrysanthemi pv. zeae in Guangdong Province%广东水稻细菌性基腐病的致病性及生物学特性研究

    Institute of Scientific and Technical Information of China (English)

    刘琼光; 曾宪铭

    1999-01-01

    通过一系列的细菌学性状和致病性试验,鉴定出广东省水稻一种新病害--水稻细菌性基腐病的病原为菊欧氏杆菌玉米致病变种Erwinia chrysanthemi pv. zeae,并对该病原菌的生物学特性进行了进一步的研究.结果表明:病菌生长的最低温度为12 ℃,适宜温度范围28~36 ℃,其中以32 ℃最适,最高温度41 ℃,致死温度53 ℃、10 min.该病菌生长的pH值范围为pH 5~11,其中pH7最适宜.

  20. Is Erwinia te beheersen? Een literatuurstudie over rotproblemen in diverse gewassen om met deze kennis Erwinia in bolgewassen beter te kunnen aanpakken

    NARCIS (Netherlands)

    Doorn, van J.; Wolf, J.

    2005-01-01

    Het meeste onderzoek aan Erwinia-bacteriën is uitgevoerd bij aardappel. In dit gewas worden drie ziekteverwekkende Erwinia-soorten gevonden: Erwinia carotovora subsp. atroseptica (Eca), Erwinia chrysanthemi (Echr) en Erwinia carotovora subsp. carotovora (Ecc). Eca speelt in gematigde klimaatsgebiede

  1. 'Candidatus Erwinia dacicola', a coevolved symbiotic bacterium of the olive fly Bactrocera oleae (Gmelin).

    Science.gov (United States)

    Capuzzo, Caterina; Firrao, Giuseppe; Mazzon, Luca; Squartini, Andrea; Girolami, Vincenzo

    2005-07-01

    The taxonomic identity of the hereditary prokaryotic symbiont of the olive fly Bactrocera oleae (Diptera: Tephritidae) was investigated. In order to avoid superficial microbial contaminants and loosely associated saprophytic biota, flies were surface-sterilized at the larval stage and reared under aseptic conditions until adult emergence. B. oleae flies originating from different geographical locations and collected at different times of the year were tested. Bacterial isolation was undertaken from the cephalic oesophageal bulb, which is known to be a specific site of accumulation for the hosted microsymbionts in the adult insect. Despite evidence of multiplication cycles taking place within the insect, attempts at cultivation of the isolated bacteria ex situ were not productive at any stage, leading to the choice of unculturable status definition. PCR amplification and nucleotide sequencing of the entire 16S rRNA gene consistently yielded a single sequence that displayed marked similarity with enterobacterial lineages, with closest matches (97%) to Erwinia persicina and Erwinia rhapontici. The novel taxon differs from common intestinal bacterial species of fruit flies and from instances of culturable bacteria previously described in B. oleae raised without sterility precautions, which we also observed as minority occupants or occasional contaminants. The symbiont's identity is also distinct from Pseudomonas savastanoi. In all observations, the numerically dominant inhabitant of the olive fly oesophageal organ was the same unculturable organism, whose presence at later stages was also regularly observed in the midgut. A novel species is proposed, by virtue of its unique properties, under the designation 'Candidatus Erwinia dacicola'.

  2. Erwinia tasmaniensis sp. nov., a non-phytopathogenic bacterium from apple and pear trees.

    Science.gov (United States)

    Geider, Klaus; Auling, Georg; Du, Zhiqiang; Jakovljevic, Vladimir; Jock, Susanne; Völksch, Beate

    2006-12-01

    Bacteria were isolated from flowers and bark of apple and pear trees at three places in Australia. In Victoria, Tasmania and Queensland, strains with white colonies on nutrient agar were screened for dome-shaped colony morphology on agar with sucrose and were found to be closely related by several criteria. The isolates were not pathogenic on apples or pears. They were characterized by a polyphasic approach including microbiological and API assays as well as fatty acid methyl ester analysis, DNA-DNA hybridization and DNA sequencing. For molecular classification, the 16S rRNA cistron and the conserved genes gpd and recA of these bacteria were investigated. Together with other taxonomic criteria, the results of these studies indicate that the bacteria belong to a novel separate species, which we propose to name Erwinia tasmaniensis sp. nov., with the type strain Et1/99(T) (=DSM 17950(T)=NCPPB 4357(T)). From DNA-DNA hybridization kinetics, microbiological characteristics and nucleotide sequence analyses, this species is related to pathogenic Erwinia species, but also to the epiphytic species Erwinia billingiae.

  3. Physiology and biochemistry of a lignin degrading bacterium Erwinia sp. Cu 3614

    Energy Technology Data Exchange (ETDEWEB)

    Rajan, J.S.

    1992-01-01

    Previous researchers have reported the isolation of a diphenylether cleaving organism, Erwinia sp., using an enrichment medium containing lignin. A copper and dinitrophenol resistant mutant of this organism, Erwinia sp. Cu3614, has also been reported. To assess the effect of copper on the growth and biochemistry of this organism, continuous cultivation was used employing an apparently optimized medium containing ethanol as carbon source. Upon increasing the concentration of copper sulfate in the medium from 5 [mu]g/ml to 10 [mu]g/ml increases in maximum specific growth rate and growth yield were seen. Also decrease in the values for doubling time and the coefficient for maintenance energy were seen. At higher levels of copper sulfate a [open quotes]non competitive[close quotes] inhibition of growth was seen as indicated by the values calculated for substrate affinity constant, and inhibition constant. To assess this organism's ligninolytic ability, an assay for residual lignin was developed. The assay measured a reaction between diazotized sulfanilic acid and lignin in alkaline solution by spectrophotometric monitoring of the resulting adduct. Use of this technique indicated that Erwinia sp. Cu3614 could degrade up to 80% of lignin in batch cultures. Further evidence about the ligninolytic ability of this organism was provided by examination of electron micrographs of lignocellulosic substrates incubated with cell suspensions. An assay for monitoring diphenylether cleaving abilities was also developed using resazurin, a redox dye. In vivo assays with cells obtained from continuous culture studies indicated a linear relationship between the rates of reaction with resazurin and the amount of copper associated with cells. In vitro assays, employing cell free extracts and resazurin, were used to obtain a fraction enriched in diphenylether cleaving activity by a heat treatment procedure.

  4. Isolation and characterization of aniline degradation slightly halophilic bacterium, Erwinia sp. Strain HSA 6.

    Science.gov (United States)

    Li, Junmin; Jin, Zexin; Yu, Binbin

    2010-07-20

    The isolated strain HSA6 is classified as Erwinia amylovora based on 16S rDNA sequence and the morphological and physiological properties. Strain HSA6 is the first reported E. amylovora in pure culture growing with aniline as sole electron donor and carbon source. The suitable pH for strain HSA6 is wide (from 5 to 11). Strain HSA6 is slightly halophilic with growth occurring at 0-10% (v/v) NaCl, and the suitable NaCl concentration for strain HSA6 is from 0% to 6%. The number of bacteria appeared to decrease with an increase in aniline concentration. The number of bacteria appeared to be constant as the wastewater concentration increased from 0% to 20%. However, the number of cells decreased with an increase in wastewater concentration from 30% to 50% and grew very slowly at 50%. The degradation rate of aniline was 100% at 0.5% aniline concentration after 24 h culture. The degradation rate of aniline was found to descend as the concentration of aniline increased from 0.5% to 3% and rose as the culture time increased. Strain HSA6 contains a plasmid with molecular weight higher than 42 kDA. Plasmid curing test and quantitative degradation test showed that strain requires the plasmid for aniline degradation. The gene cluster degrading aniline was determined in the plasmid by PCR amplification.

  5. The antimicrobial activity of chitosan on the pathogen (Erwinia chrysanthemi pv.Zeae) of bacterial stalk rot of corn determined by colorimetry%比色法测定壳聚糖对玉米细菌性茎腐病菌的抑制作用

    Institute of Scientific and Technical Information of China (English)

    彭晓江; 刘晓津; 邱道寿; 曾粮斌

    2004-01-01

    通过选择合适的波长建立了玉米细菌性茎腐病菌(Erwinia chrysanthemi pv.zeae)悬浮液浓度与吸光值间的标准曲线,在LB液体培养基中分别设置6种壳聚糖浓度(0、0.5、1.0、2.0、3.5、5.5 mg/mL),用比色分析法测定了各浓度下玉米茎腐病菌的生长状况,并建立生长曲线.通过生长曲线分析表明壳聚糖对玉米茎腐病菌具有显著的抑制作用,且作用强度随壳聚糖浓度的提高而增强.

  6. CLONIG AND CHARACTERIZATION OF A NEW ICE NUCLEATION ACTIVEGENE FROM ICE NUCLEATION ACTIVE BACTERIUM ERWINIA ANANAS 110%冰核细菌(Erwinia ananas 110)冰核基因克隆和序列分析

    Institute of Scientific and Technical Information of China (English)

    唐朝荣; 孙福在; 赵廷昌; 李瑞峰

    2002-01-01

    从作者自行分离的冰核细菌(Erwinia ananas110)中克隆到我国第1个细菌冰核基因,并完成其序列测定和分析.所克隆基因编码区全长3921bp,编码1306aa,氨基酸序列明显分为3个区即N-端(161aa)、C-端(41aa)的单一序列区和中部的高度重复序列R区(1104aa),以16氨基酸为重复单元的R区占整个编码序列的84.5%.序列分析表明我们所克隆的基因为一个新冰核基因,将其命名为iceA,该基因已在GenBank上登录,登录号为:AF387802.

  7. The alternative sigma factor HrpL negatively modulates the flagellar system in the phytopathogenic bacterium Erwinia amylovora under hrp-inducing conditions.

    Science.gov (United States)

    Cesbron, Sophie; Paulin, Jean-Pierre; Tharaud, Michel; Barny, Marie-Anne; Brisset, Marie-Noëlle

    2006-04-01

    In this work we present evidence of an opposite regulation in the phytopathogenic bacteria Erwinia amylovora between the virulence-associated Type III secretion system (TTSS) and the flagellar system. Using loss-of-function mutants we show that motility enhanced the virulence of wild-type bacteria relative to a nonmotile mutant when sprayed on apple seedlings with unwounded leaves. Then we demonstrated through analyses of motility, flagellin export and visualization of flagellar filament that HrpL, the positive key regulator of the TTSS, also down-regulates the flagellar system. Such a dual regulation mediated by an alternative sigma factor of the TTSS appears to be a level of regulation between virulence and motility not yet described among Proteobacteria.

  8. Expression of Exogenous Gene in Bacterium Erwinia uredovora%外源基因在噬夏孢欧文氏菌中的表达

    Institute of Scientific and Technical Information of China (English)

    汪靖超; 赵驰; 王波; 杨宏; 李荣贵

    2005-01-01

    用CaCl2法制备噬夏孢欧文氏菌(Erwinia uredovora)感受态细胞,热激处理可使质粒pACYC184转入噬夏孢欧文氏菌细胞,转化率为656 cfu/μg DNA,转入的质粒可以在细胞中稳定存在,并随着细胞分裂而复制,质粒上的氯霉素抗性基因在其启动子的控制下能够高效表达,氯霉素乙酰转移酶可达噬夏孢欧文氏菌菌体总蛋白的30.26%.噬夏孢欧文氏菌可以作为一种新的原核表达系统.

  9. Isolation and Identification of L-asparaginase producing Erwinia strains which isolated from Potato Farms

    Directory of Open Access Journals (Sweden)

    Arastoo Badoei-Dalfard

    2016-09-01

    Full Text Available Introduction: L-Asparaginase can be effectively used for the treatment of lymphoblastic leukemia. The rapid growth of cancer cells are needed for L-asparagine abundant storage. L-asparaginase catalyzes the hydrolysis of L-asparagine into L-aspartic acid and ammonia. The purpose of this study was to isolate and identify the L-asparaginase producing Erwinia strains from the potato farms of Jiroft. Materials and methods: Pectolytic Erwinia species isolated from crumbling potato in M9 medium. The desired L-asparaginase producing bacteria were isolated based on the color changes. Biochemical-microbial and the plant pathogenicity tests of these strains were also investigated with potato and geranium. The L-asparaginase production and molecular detection of these Erwinia strains were also investigated. Results: In this study, L-asparaginase producing Erwinia was isolated on the CVP and M9 mediums. The inoculation of Erwinia strains on the potato and geranium plants showed that Er8 and Er11 species have the ability to cause plant pathogenicity. Results showed that the maximum pathogenicity of Er8 and Er11 was observed after 48 and 15 h of inoculation in potato and geranium plants, respectively. 16S rDNA sequencing and phylogenetic analyses exhibited that Er8 and Er11 strains were similar to Erwinia chrysanthemi with 98% homology. Discussion and conclusion: Because of several applications of the Erwinia L-asparaginase in various fields, isolated Erwinia and their L-asparaginase can be suitable for applied utilization.

  10. Bacterieziekte Erwinia groeiend probleem

    NARCIS (Netherlands)

    Wolf, van der J.M.

    2012-01-01

    Het grootste probleem van Nederlandse pootgoedtelers is tegenwoordig de bacterieziekte Erwinia. Het is een sluipmoordenaar waar nog geen bestrijdingsmiddelen tegen bestaan. Maar onderzoekers komen steeds meer over de bacterie te weten.

  11. Hypersensitive response and acyl-homoserine lactone production of the fire blight antagonists Erwinia tasmaniensis and Erwinia billingiae.

    Science.gov (United States)

    Jakovljevic, Vladimir; Jock, Susanne; Du, Zhiqiang; Geider, Klaus

    2008-09-01

    Fire blight caused by the Gram-negative bacterium Erwinia amylovora can be controlled by antagonistic microorganisms. We characterized epiphytic bacteria isolated from healthy apple and pear trees in Australia, named Erwinia tasmaniensis, and the epiphytic bacterium Erwinia billingiae from England for physiological properties, interaction with plants and interference with growth of E. amylovora. They reduced symptom formation by the fire blight pathogen on immature pears and the colonization of apple flowers. In contrast to E. billingiae, E. tasmaniensis strains induced a hypersensitive response in tobacco leaves and synthesized levan in the presence of sucrose. With consensus primers deduced from lsc as well as hrpL, hrcC and hrcR of the hrp region of E. amylovora and of related bacteria, these genes were successfully amplified from E. tasmaniensis DNA and alignment of the encoded proteins to other Erwinia species supported a role for environmental fitness of the epiphytic bacterium. Unlike E. tasmaniensis, the epiphytic bacterium E. billingiae produced an acyl-homoserine lactone for bacterial cell-to-cell communication. Their competition with the growth of E. amylovora may be involved in controlling fire blight.

  12. L-asparaginase treatment in acute lymphoblastic leukemia: a focus on Erwinia asparaginase

    Science.gov (United States)

    Pieters, Rob; Hunger, Stephen P; Boos, Joachim; Rizzari, Carmelo; Silverman, Lewis; Baruchel, Andre; Goekbuget, Nicola; Schrappe, Martin; Pui, Ching-Hon

    2010-01-01

    Asparaginases are a cornerstone of treatment protocols for acute lymphoblastic leukemia (ALL) and are used for remission induction and intensification treatment in all pediatric regimens and in the majority of adult protocols. Extensive clinical data have shown that intensive asparaginase treatment improves clinical outcomes in childhood ALL. Three asparaginase preparations are available; the native asparaginase derived from Escherichia coli (E. coli-asparaginase), a pegylated form of this enzyme (PEG-asparaginase) and a product isolated from Erwinia chrysanthemi, i.e. Erwinia asparaginase. Clinical hypersensitivity reactions and silent inactivation due to antibodies against E.coli-asparaginase, lead to inactivation of E-Coli asparaginase in up to 60% of cases. Current treatment protocols include E. coli-asparaginase or PEG-asparaginase for first-line treatment of ALL. Typically, patients exhibiting sensitivity to one formulation of asparaginase are switched to another product to ensure they receive the most efficacious treatment regimen possible. Erwinia asparaginase is used as a second- or third-line treatment in European and US protocols. Despite the universal inclusion of asparaginase in such treatment protocols, there is much debate regarding the optimal formulation and dosage of these agents. This manuscript provides an overview of available evidence to make recommendations for optimal use of Erwinia asparaginase in the treatment of ALL. PMID:20824725

  13. L-asparaginase treatment in acute lymphoblastic leukemia: a focus on Erwinia asparaginase.

    Science.gov (United States)

    Pieters, Rob; Hunger, Stephen P; Boos, Joachim; Rizzari, Carmelo; Silverman, Lewis; Baruchel, Andre; Goekbuget, Nicola; Schrappe, Martin; Pui, Ching-Hon

    2011-01-15

    Asparaginases are a cornerstone of treatment protocols for acute lymphoblastic leukemia (ALL) and are used for remission induction and intensification treatment in all pediatric regimens and in the majority of adult treatment protocols. Extensive clinical data have shown that intensive asparaginase treatment improves clinical outcomes in childhood ALL. Three asparaginase preparations are available: the native asparaginase derived from Escherichia coli (E. coli asparaginase), a pegylated form of this enzyme (PEG-asparaginase), and a product isolated from Erwinia chrysanthemi, ie, Erwinia asparaginase. Clinical hypersensitivity reactions and silent inactivation due to antibodies against E. coli asparaginase, lead to inactivation of E. coli asparaginase in up to 60% of cases. Current treatment protocols include E. coli asparaginase or PEG-asparaginase for first-line treatment of ALL. Typically, patients exhibiting sensitivity to one formulation of asparaginase are switched to another to ensure they receive the most efficacious treatment regimen possible. Erwinia asparaginase is used as a second- or third-line treatment in European and US protocols. Despite the universal inclusion of asparaginase in such treatment protocols, debate on the optimal formulation and dosage of these agents continues. This article provides an overview of available evidence for optimal use of Erwinia asparaginase in the treatment of ALL.

  14. Monitoring of Erwinia amylovora in Montenegro

    Directory of Open Access Journals (Sweden)

    Dragana Radunović

    2015-09-01

    Full Text Available Recent studies of Erwinia amylovora in Montenegro, conducted from 2012 to 2014, indicated that the bacterium was widespread in the northern, continental part of the country, where the most important fruit-growing regions are situated. The presence of the bacterium was confirmed on quince, pear, apple, medlar and hawthorn. Pathogenic, cultural and biochemical characteristics of E. amylovora strains sampled from pome fruit species and indigenous flora in Montenegro had been studied previously. In the present study, serological tests were used for identification of E. amylovora strains originating from pome fruit trees and indigenous plants. Monitoring of E. amylovora and collection of samples with symptoms of bacterial fire blight from different hosts and locations were performed in Montenegro from 2012 to 2014. Isolation of the bacterium on nutrient medium produced a large number of isolates, whose pathogenicity was confirmed on immature pear fruits. Twenty-seven strains of the bacterium, originating from three pome fruit species (quince, pear and apple and one indigenous species (hawthorn were selected for serological analyses. Two applied serological methods, ELISA and IF test, enabled rapid detection of the bacterium and simultaneous examination of a large number of samples over a short period of time. Serological analyses showed high homogeneity in antigenic structure of the studied E. amylovora strains sampled from quince, pear, apple and hawthorn from nine locations in Montenegro.

  15. 'Preventie belangrijkste troef tegen Erwinia'

    NARCIS (Netherlands)

    Doorn, van J.

    2012-01-01

    De bollenteelt en -handel ondervindt aanzienlijke schade van de bacterieziekte Erwinia. Onderzoek wijst uit dat preventie het belangrijkste wapen is. Mogelijk bieden ook stofjes die de afweer van planten verbeteren een oplossing.

  16. Dickeya dadantii, a plant pathogenic bacterium producing Cyt-like entomotoxins, causes septicemia in the pea aphid Acyrthosiphon pisum.

    Directory of Open Access Journals (Sweden)

    Denis Costechareyre

    Full Text Available Dickeya dadantii (syn. Erwinia chrysanthemi is a plant pathogenic bacteria that harbours a cluster of four horizontally-transferred, insect-specific toxin genes. It was recently shown to be capable of causing an acute infection in the pea aphid Acyrthosiphon pisum (Insecta: Hemiptera. The infection route of the pathogen, and the role and in vivo expression pattern of these toxins, remain unknown. Using bacterial numeration and immunolocalization, we investigated the kinetics and the pattern of infection of this phytopathogenic bacterium within its insect host. We compared infection by the wild-type strain and by the Cyt toxin-deficient mutant. D. dadantii was found to form dense clusters in many luminal parts of the aphid intestinal tract, including the stomach, from which it invaded internal tissues as early as day 1 post-infection. Septicemia occurred soon after, with the fat body being the main infected tissue, together with numerous early infections of the embryonic chains showing embryonic gut and fat body as the target organs. Generalized septicemia led to insect death when the bacterial load reached about 10(8 cfu. Some individual aphids regularly escaped infection, indicating an effective partial immune response to this bacteria. Cyt-defective mutants killed insects more slowly but were capable of localisation in any type of tissue. Cyt toxin expression appeared to be restricted to the digestive tract where it probably assisted in crossing over the first cell barrier and, thus, accelerating bacterial diffusion into the aphid haemocel. Finally, the presence of bacteria on the surface of leaves hosting infected aphids indicated that the insects could be vectors of the bacteria.

  17. Toetsen om Erwinia in bloembollen aan te tonen : onderzoek Erwinia

    NARCIS (Netherlands)

    Doorn, van J.; Hollinger, T.C.; Kampen, van D.; Vreeburg, P.J.M.; Leeuwen, van P.J.; Wolf, van der J.M.

    2007-01-01

    Binnen het Erwiniaproject is een aantal toetsen ontwikkeld en zijn andere nog in ontwikkeling. Deze toetsen moeten liefst ook latent aanwezige Erwiniastammen kunnen aantonen. Dit lukt door monsters in een voedingsbodem voor Erwinia te brengen en na kweek deze bacteriën aan te tonen met serologische

  18. Simplified Extraction and Cleanup for Multiresidue Determination of Pyrethroid Insecticides in Chrysanthemi

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    A simplified method for determining 4 pyrethroid insecticides (Fenpropathrin,Cypermethrin,Fenvalirate,and Deltamethrin)in Chrysanthemi is described.Standards were fortified in to Chrysanthemi (5g)with two levels,)0.1-1ppm the pyrethroid pesticides are extracted with petroleum ether and cleaned up by natural aluminum oxide.The extracts are analyzed by gas chromatography equipped with electron capture detector (ECD).Analysis of fortified Chrysanthemi(n=3)shows recovery range from 99-102% at 1 ppm level and 93-104% at 0.1 ppm level. The coefficient of variation of the method at 1st level was 0.12 and 0.14 for 2ed level.The detection limits of the method ranged from 0.04ppb to 28ppb.The method is repid,sample,and sensitive and is applicable to the determination of Fenpropathrin,Cypermethrin,Fenvalirate,and Deltamethrin in Chrysanthemi.

  19. The hexA gene of Erwinia carotovora encodes a LysR homologue and regulates motility and the expression of multiple virulence determinants.

    Science.gov (United States)

    Harris, S J; Shih, Y L; Bentley, S D; Salmond, G P

    1998-05-01

    We have identified a gene important for the regulation of exoenzyme virulence factor synthesis in the plant pathogen Erwinia carotovora ssp. carotovora (Ecc) and virulence and motility in Erwinia carotovora ssp. atroseptica (Eca). This gene, hexA (hyperproduction of exoenzymes), is a close relative of the Erwinia chrysanthemi (Echr) gene pecT and encodes a member of the LysR family of transcriptional regulators. hexA mutants in both Ecc and Eca produce abnormally high levels of the exoenzyme virulence factors pectate lyase, cellulase and protease. In addition, Eca hexA mutants show increased expression of the fliA and fliC genes and hypermotility. Consistent with a role as a global regulator, expression of hexA from even a low-copy plasmid can suppress exoenzyme production in Ecc and Eca and motility in Eca. Production of the quorum-sensing pheromone OHHL in Ecc hexA is higher throughout the growth curve compared with the wild-type strain. Overexpression of Ecc hexA also caused widespread effects in several strains of the opportunistic human pathogen, Serratia. Low-copy hexA expression resulted in repression of exoenzyme, pigment and antibiotic production and repression of the spreading phenotype. Finally, mutations in hexA were shown to increase Ecc or Eca virulence in planta.

  20. Overleving van Erwinia in water : waakzaamheid geboden

    NARCIS (Netherlands)

    Doorn, van J.; Houtenbos, R.; Wolf, van der J.M.

    2008-01-01

    PPO en Plant Research International hebben al eerder de rotbacterie Erwinia onderzocht, omdat Erwinia een vochtminnende bacterie is, is besmetting via water, ook vanuit de lucht een reëel gevaar. Omdat water op veel manieren op bollenbedrijven toegepast wordt is onderzocht hoe lang deze rotveroorzak

  1. Determination of camphor and borneol in Flos Chrysanthemi Indici by UAE and GC-FID.

    Science.gov (United States)

    Ye, Qing; Deng, Chunhui

    2009-04-01

    In the work, ultrasonic-assisted extraction (UAE) followed by gas chromatography with flame ionization detector (GC-FID) is developed for the quantitative analysis of the bioactive components of camphor and borneol in a traditional Chinese medicine (TCM) of Flos Chrysanthemi Indici. The extraction parameters are investigated. The optimum extraction conditions found are: solvent, methanol; solvent to sample ratio, 12:1 (v/w); extraction time, 15 min. Camphor and borneol are determined using this extraction method in Flos Chrysanthemi Indici samples from 5 different growing areas. The relative standard deviation values for camphor and borneol are 8.4% and 5.6%, respectively. The recoveries for camphor and borneol are 89% and 95%, respectively, and the method detection limits are lower than 0.23 microg/mL. To demonstrate the method feasibility, steam distillation is also used to analyze camphor and borneol in Flos Chrysanthemi Indici samples from these different growing areas. The statistical comparison by t-test (95% confidence level) showed no significant difference between these results. It has been shown that the proposed UAE-GC-FID is a simple, rapid, and reliable method for quantitative analysis of camphor and borneol in TCM, and a potential tool for quality assessment of Flos Chrysanthemi Indici.

  2. Erwinia berokkent pootgoedsector vele miljoenen schade

    NARCIS (Netherlands)

    Prins, H.; Breukers, A.

    2008-01-01

    De besmetting met Erwinia ontwikkelt zich de laatste jaren tot een serieus probleem in de pootgoedsector. Voor telers ontstaat inkomensderving door lagere opbrengstprijzen en onbruikbare pootgoedvoorraden. Handelshuizen lijden schade door margedaling en reclameringen. Een aantasting van het imago va

  3. The influence of plant extracts on growth of Erwinia amylovora - the causal agent of fire blight

    Directory of Open Access Journals (Sweden)

    Grzegorz Krupiński

    2013-12-01

    Full Text Available Ethanol and water extracts obtained on Soxhlet apparatus from various organs and parts (leaves, flowers, shoots, onion, bark, fruit of 30 herbal and woody plants species were tested for growth inhibition of Erwinia amylovora using agar diffusion method. Active extracts were found in 23 plant species but in 13 ofthem it was found for the first time. The highest diameter of growth inhibition zone of this bacterium was caused by extracts from Aloe arborescens, Juglans regia, Rhus typhina, Salvia offici nalis and Satureja hortensis. In almost all cases ethanol appeared to be a better solvent of active plant substances against E.amylovora than water.

  4. Bacteria of the genus Erwinia found in the spermatheca of the laurel psyllid Trioza alacris.

    Science.gov (United States)

    Marchini, Daniela; Ciolfi, Silvia; Gottardo, Marco; Marri, Laura

    2014-12-01

    Psylloidea are economically important insects causing serious damage to plants by direct feeding and/or vectoring bacterial pathogens. Results reported here indicate the presence of extracellular bacteria in the spermatheca of egg-laying Trioza alacris females. One phylotype, sharing 99 % identity with the non-phytopathogenic bacterium Erwinia tasmaniensis, was identified regardless of methods applied or insect sampling year and location. This is the first study, achieved by ultrastructural, cultural, and 16S rRNA gene-based analysis, of an insect spermatheca microbiota.

  5. Carbohydrate and ethane release with Erwinia carotovora subspecies betavasculorum--induced necrosis.

    Science.gov (United States)

    Kuykendall, L David; Hunter, William J

    2008-02-01

    Erwinia carotovora subspecies betavasculorum, also known as E. betavasculorum and Pectobacterium betavasculorum, is a soil bacterium that has the capacity to cause root rot necrosis of sugarbeets. The qualitatively different pathogenicity exhibited by the virulent E. carotovora strain and two avirulent strains, a Citrobacter sp. and an Enterobacter cloacae, was examined using digital analysis of photographic evidence of necrosis as well as for carbohydrate, ethane, and ethylene release compared with uninoculated potato tuber slices. Visual scoring of necrosis was superior to digital analysis of photographs. The release of carbohydrates and ethane from potato tuber slices inoculated with the soft rot necrosis-causing Erwinia was significantly greater than that of potato tuber slices that had not been inoculated or that had been inoculated with the nonpathogenic E. cloacae and Citrobacter sp. strains. Interestingly, ethylene production from potato slices left uninoculated or inoculated with the nonpathogenic Citrobacter strain was 5- to 10-fold higher than with potato slices inoculated with the pathogenic Erwinia strain. These findings suggest that (1) carbohydrate release might be a useful measure of the degree of pathogenesis, or relative virulence; and that (2) bacterial suppression of ethylene formation may be a critical step in root rot disease formation.

  6. Functional Characterization of Cyclic Adenosine Monphosphate (cAMP)Recptor Protein Gene (crp) from Erwinia amylovora%梨火疫菌(Erwinia amylovora)环腺苷酸受体蛋白基因(crp)的功能分析

    Institute of Scientific and Technical Information of China (English)

    刘倩倩; 于洋洋; 宋俊贤; 胡白石; 范加勤; 刘凤权

    2010-01-01

    梨火疫菌(Erwinia amylovora)可引起梨、苹果等蔷薇科(Rosaceae)植物的火疫病.在菊欧文氏菌(Erwinia chrysanthemi)中,由crp基因编码的环腺苷酸受体蛋白(cyclic adenosine monphosphate(cAMP)receptor protein,CRP)对果胶酶基因的表达调控和菌株致病性起着重要的作用.本研究首次鉴定并克隆出梨火疫菌中的crp同源基因,命名为Eacrp,并通过同源重组的方法,构建了梨火疫菌的crp基因突变体Ea△crp以及互补子,进行了致病性、过敏性反应、胞外多糖、鞭毛运动等一系列相关表型的鉴定.结果表明,crp基因影响着梨火疫菌的致病性、胞外多糖、游动性、生长情况等多种生物学特性,然而,Ea△crp仍能引起烟草过敏性反应,并且在过氧化氢敏感度以及沉降性、生物膜和AI-2信号分子的生成方面与野生型菌株相比差异明显.本研究结果说明,梨火疫菌crp基因对病菌的胞外多糖分泌、生长、游动性以及致病性方面具有关键作用.

  7. Export of the HR eliciting protein, Harpin(Es), of the maize pathogen Erwinia stewartii is species-specific but is independent of the growth temperature.

    Science.gov (United States)

    Ahmad, Musharaf; Alam, Syed Sartaj; Alam, Shah; Usman, Amjad; Coplin, David L

    2007-01-01

    The extra-cellular export of the HR-eliciting protein, Harpin(Es) of the maize pathogen Erwinia stewartii was studied to find out if the protein needs any species-specific signal for its export and to determine if the export of the protein to the medium is affected in any way by the growth temperature. Based upon the experimental evidence, it was proved that the protein (i.e., Harpin(Es)) does require its own export system (species-specific) to get out of the bacterial cell and can not be exported by the export system of even the very closely related bacterium, Erwinia amylovora. It was also found that the export of Harpin(Es) is, unlike the case of Harpin(Ea) (HR-eliciting protein of Erwinia amylovora), independent of the growth temperature.

  8. Comparative genomics of Japanese Erwinia pyrifoliae strain Ejp617 with closely related erwinias.

    Science.gov (United States)

    Thapa, Shree P; Park, Duck H; Kim, Won S; Choi, Beom S; Lim, Jong S; Choi, Ik Y; Hur, Jang H; Lim, Chun K

    2013-02-01

    Japanese Erwinia pyrifoliae strains cause bacterial shoot blight of pear (BSBP) in Japan. The genetics of Japanese Erwinia remains largely unknown relative to the abundant genomic information available for other Erwinia strains. We compared the genome of Japanese and Korean E. pyrifoliae strains along with those of E. amylovora and E. tasmaniensis. Comparisons with the Korean E. pyrifoliae strain revealed numerous gene insertions/deletions, rearrangements, and inversions in the central regions of the chromosomes. Approximately 80% (2843) of coding DNA sequences (CDSs) are shared by these two genomes which represent about three-quarters of the genome, and there are about 20% unique CDSs. Comparative analysis with closely related erwinias showed that 1942 (more than 50%) core open reading frames (ORF) are shared by all these strains. In addition to two type III secretion systems (hrp/dsp and inv/spa), the genome of Ejp617 encodes numerous virulence factors, including a type VI secretion system, an exopolysaccharide synthesis cluster, and another protein secretion system present in plant pathogenic Erwinia strains. The availability of whole genome sequence should provide a resource to further improve the understanding of pathogenesis in Japanese E. pyrifoliae Ejp617 and to facilitate evolutionary studies among the species of the genus Erwinia.

  9. Evolutionary insights from Erwinia amylovora genomics.

    Science.gov (United States)

    Smits, Theo H M; Rezzonico, Fabio; Duffy, Brion

    2011-08-20

    Evolutionary genomics is coming into focus with the recent availability of complete sequences for many bacterial species. A hypothesis on the evolution of virulence factors in the plant pathogen Erwinia amylovora, the causative agent of fire blight, was generated using comparative genomics with the genomes E. amylovora, Erwinia pyrifoliae and Erwinia tasmaniensis. Putative virulence factors were mapped to the proposed genealogy of the genus Erwinia that is based on phylogenetic and genomic data. Ancestral origin of several virulence factors was identified, including levan biosynthesis, sorbitol metabolism, three T3SS and two T6SS. Other factors appeared to have been acquired after divergence of pathogenic species, including a second flagellar gene and two glycosyltransferases involved in amylovoran biosynthesis. E. amylovora singletons include 3 unique T3SS effectors that may explain differential virulence/host ranges. E. amylovora also has a unique T1SS export system, and a unique third T6SS gene cluster. Genetic analysis revealed signatures of foreign DNA suggesting that horizontal gene transfer is responsible for some of these differential features between the three species.

  10. Trefkans detectie Erwinia in pootgoed toegenomen

    NARCIS (Netherlands)

    Wolf, van der J.M.

    2014-01-01

    Binnen het Deltaplan Erwinia is een methode ontwikkeld die met grotere zekerheid de aanwezigheid van de Erwinia’s, Pectobacterium en Dickeya, in pootgoed kan aantonen. De verbeterde methode maakt gebruik gemaakt van uitschotknollen die op de sorteerband uitgeselecteerd zijn en een toets waarbij de g

  11. Deltaplan Erwinia hielp piepers en bollen vooruit

    NARCIS (Netherlands)

    Dwarswaard, A.; Doorn, van J.

    2013-01-01

    Vier jaar lang werkte de bloembollensector samen met de aardappelsector aan de gezamenlijke bacterieziekte Erwinia. In beide sectoren was al het nodige voorwerk gedaan. Op 12 december 2012 werd het project afgesloten. Met de hoop op toch nog enige vorm van voortzetting.

  12. New Erwinia-like organism causing cervical lymphadenitis.

    Science.gov (United States)

    Shin, Sang Yop; Lee, Mi Young; Song, Jae-Hoon; Ko, Kwan Soo

    2008-09-01

    The first case of cervical lymphadenitis due to infection by a new Erwinia-like organism is reported. The organism was identified initially as Pantoea sp. by a Vitek 2-based assessment but was finally identified as a member of the genus Erwinia by 16S rRNA gene sequence analysis. The isolate displayed 98.9% 16S rRNA gene sequence similarity to that of E. tasmaniensis and showed phenotypic characteristics that were different from other Erwinia species.

  13. Expression of lysozymes from Erwinia amylovora phages and Erwinia genomes and inhibition by a bacterial protein.

    Science.gov (United States)

    Müller, Ina; Gernold, Marina; Schneider, Bernd; Geider, Klaus

    2012-01-01

    Genes coding for lysozyme-inhibiting proteins (Ivy) were cloned from the chromosomes of the plant pathogens Erwinia amylovora and Erwinia pyrifoliae. The product interfered not only with activity of hen egg white lysozyme, but also with an enzyme from E. amylovora phage ΦEa1h. We have expressed lysozyme genes from the genomes of three Erwinia species in Escherichia coli. The lysozymes expressed from genes of the E. amylovora phages ΦEa104 and ΦEa116, Erwinia chromosomes and Arabidopsis thaliana were not affected by Ivy. The enzyme from bacteriophage ΦEa1h was fused at the N- or C-terminus to other peptides. Compared to the intact lysozyme, a His-tag reduced its lytic activity about 10-fold and larger fusion proteins abolished activity completely. Specific protease cleavage restored lysozyme activity of a GST-fusion. The bacteriophage-encoded lysozymes were more active than the enzymes from bacterial chromosomes. Viral lyz genes were inserted into a broad-host range vector, and transfer to E. amylovora inhibited cell growth. Inserted in the yeast Pichia pastoris, the ΦEa1h-lysozyme was secreted and also inhibited by Ivy. Here we describe expression of unrelated cloned 'silent' lyz genes from Erwinia chromosomes and a novel interference of bacterial Ivy proteins with a viral lysozyme.

  14. A multiplex real-time PCR assay for the detection of Puccinia horiana and P. chrysanthemi on chrysanthemum

    Science.gov (United States)

    Puccinia horiana, the cause of chrysanthemum white rust, is a regulated fungal plant pathogen in the United States, while Puccinia chrysanthemi, the cause of chrysanthemum brown rust, is a widespread but less destructive pathogen. Accurate identification of these pathogens is essential to correctly ...

  15. Evaluation of Susceptibility of Different Pear Hybrid Populations to Fire Blight (Erwinia amylovora

    Directory of Open Access Journals (Sweden)

    Yasemin EVRENOSOĞLU

    2011-05-01

    Full Text Available Fire blight disease caused by pathogenic bacterium Erwinia amylovora, is the serious disease of pear, and there is not a certain chemical management against this disease except antibiotic-type compounds such as streptomycin. It is very important to improve new fire blight resistant cultivars in case of integrated disease management. With this purpose, different crosses have been made between Pyrus communis varieties that have good fruit characteristics and resistant cultigens. Besides, self and open pollination treatments have been carried out in maternal plants. The disease resistance level of the hybrids obtained from these combinations was determined by artificial inoculations by Erwinia amylovora in greenhouse conditions. A total of 3284 hybrids were inoculated, and 2631 of them survived and were distributed to different susceptibility classes. 19.88% of the inoculated hybrids was killed by Erwinia amylovora. Total distribution of the hybrids to susceptibility classes was as 6.18% in class “A- slightly susceptible”, 3.11% in class “B- less susceptible”, 8.89% in class “C- mid-susceptible”, 20.28% in class “D- susceptible”, and 61.54% in class “E- very susceptible”. Majority of class “A- slightly susceptible” hybrids were obtained from ‘Magness’ x ‘Ankara’ combination. ‘Kieffer’ x ‘Santa Maria’, ‘Kieffer’ open pollination, ‘Magness’ x ‘Akça’, ‘Magness’ x ‘Kieffer’, ‘Magness’ x ‘Santa Maria’, ‘Mustafa Bey’ x ‘Moonglow’ treatments displayed good results with respect to “A- slightly susceptible” character. It is very important to evaluate these hybrid pear populations through different fruit and tree characteristics in the future.

  16. Erwinia chrysanthemi Burkholder, McFadden and Dimock and other Phytobacteria Causal Agents of Onion (Allium cepa L.) Bulb Decay, and their Detection

    OpenAIRE

    María de Jesús Yáñez; Leopold Fucikovsky; James W. Lorbeer; Armando González; Sergio Aranda

    2003-01-01

    For the first time, a severe soft rot of onion bulbs in commercial fields was detected in southern Tamaulipas and eastern San Luis Potosi States, Mexico, in 1989. To identify the causal agent, 50 rotten bulbs were analyzed. Commercial seed lots, soil, and irrigation water were also investigated as a potential sources of inoculum. Pathogenic bacteria isolated from bulbs were identified by the biolog microplate system (BMS), Ouchterlony double diffusion serology, physiological tests, and only b...

  17. Complete genome sequence of the fire blight pathogen Erwinia amylovora CFBP 1430 and comparison to other Erwinia spp.

    Science.gov (United States)

    Smits, Theo H M; Rezzonico, Fabio; Kamber, Tim; Blom, Jochen; Goesmann, Alexander; Frey, Jürg E; Duffy, Brion

    2010-04-01

    Fire blight, caused by the enterobacterium Erwinia amylovora, is a devastating disease of rosaceous plants that has global economic importance for apple and pear production and trade. The complete genome of E. amylovora CFBP 1430 was sequenced, annotated, and compared with the genomes of other Erwinia spp. Several singleton and shared features of the E. amylovora CFBP 1430 genome were identified that offer a first view into evolutionary aspects within the genus Erwinia. Comparative genomics identified or clarified virulence and fitness determinants and secretion systems. Novel insights revealed in the genome of E. amylovora CFBP 1430 hold potential for exploitation to improve the design of more effective fire blight control strategies.

  18. Comparative Genomics of Erwinia amylovora and Related Erwinia Species—What do We Learn?

    Directory of Open Access Journals (Sweden)

    Youfu Zhao

    2011-09-01

    Full Text Available Erwinia amylovora, the causal agent of fire blight disease of apples and pears, is one of the most important plant bacterial pathogens with worldwide economic significance. Recent reports on the complete or draft genome sequences of four species in the genus Erwinia, including E. amylovora, E. pyrifoliae, E. tasmaniensis, and E. billingiae, have provided us near complete genetic information about this pathogen and its closely-related species. This review describes in silico subtractive hybridization-based comparative genomic analyses of eight genomes currently available, and highlights what we have learned from these comparative analyses, as well as genetic and functional genomic studies. Sequence analyses reinforce the assumption that E. amylovora is a relatively homogeneous species and support the current classification scheme of E. amylovora and its related species. The potential evolutionary origin of these Erwinia species is also proposed. The current understanding of the pathogen, its virulence mechanism and host specificity from genome sequencing data is summarized. Future research directions are also suggested.

  19. Mutational analysis of a predicted double β-propeller domain of the DspA/E effector of Erwinia amylovora.

    Science.gov (United States)

    Siamer, Sabrina; Gaubert, Stéphane; Boureau, Tristan; Brisset, Marie-Noëlle; Barny, Marie-Anne

    2013-05-01

    The bacterium Erwinia amylovora causes fire blight, an invasive disease that threatens apple trees, pear trees and other plants of the Rosaceae family. Erwinia amylovora pathogenicity relies on a type III secretion system and on a single effector DspA/E. This effector belongs to the widespread AvrE family of effectors whose biological function is unknown. In this manuscript, we performed a bioinformatic analysis of DspA/E- and AvrE-related effectors. Motif search identified nuclear localization signals, peroxisome targeting signals, endoplasmic reticulum membrane retention signals and leucine zipper motifs, but none of these motifs were present in all the AvrE-related effectors analysed. Protein threading analysis, however, predicted a conserved double β-propeller domain in the N-terminal part of all the analysed effector sequences. We then performed a random pentapeptide mutagenesis of DspA/E, which led to the characterization of 13 new altered proteins with a five amino acids insertion. Eight harboured the insertion inside the predicted β-propeller domain and six of these eight insertions impaired DspA/E stability or function. Conversely, the two remaining insertions generated proteins that were functional and abundantly secreted in the supernatant suggesting that these two insertions stabilized the protein.

  20. Erwinia amylovora novel plasmid pEI70: complete sequence, biogeography, and role in aggressiveness in the fire blight phytopathogen.

    Directory of Open Access Journals (Sweden)

    Pablo Llop

    Full Text Available Comparative genomics of several strains of Erwinia amylovora, a plant pathogenic bacterium causal agent of fire blight disease, revealed that its diversity is primarily attributable to the flexible genome comprised of plasmids. We recently identified and sequenced in full a novel 65.8 kb plasmid, called pEI70. Annotation revealed a lack of known virulence-related genes, but found evidence for a unique integrative conjugative element related to that of other plant and human pathogens. Comparative analyses using BLASTN showed that pEI70 is almost entirely included in plasmid pEB102 from E. billingiae, an epiphytic Erwinia of pome fruits, with sequence identities superior to 98%. A duplex PCR assay was developed to survey the prevalence of plasmid pEI70 and also that of pEA29, which had previously been described in several E. amylovora strains. Plasmid pEI70 was found widely dispersed across Europe with frequencies of 5-92%, but it was absent in E. amylovora analyzed populations from outside of Europe. Restriction analysis and hybridization demonstrated that this plasmid was identical in at least 13 strains. Curing E. amylovora strains of pEI70 reduced their aggressiveness on pear, and introducing pEI70 into low-aggressiveness strains lacking this plasmid increased symptoms development in this host. Discovery of this novel plasmid offers new insights into the biogeography, evolution and virulence determinants in E. amylovora.

  1. Erwinia amylovora novel plasmid pEI70: complete sequence, biogeography, and role in aggressiveness in the fire blight phytopathogen.

    Science.gov (United States)

    Llop, Pablo; Cabrefiga, Jordi; Smits, Theo H M; Dreo, Tanja; Barbé, Silvia; Pulawska, Joanna; Bultreys, Alain; Blom, Jochen; Duffy, Brion; Montesinos, Emilio; López, María M

    2011-01-01

    Comparative genomics of several strains of Erwinia amylovora, a plant pathogenic bacterium causal agent of fire blight disease, revealed that its diversity is primarily attributable to the flexible genome comprised of plasmids. We recently identified and sequenced in full a novel 65.8 kb plasmid, called pEI70. Annotation revealed a lack of known virulence-related genes, but found evidence for a unique integrative conjugative element related to that of other plant and human pathogens. Comparative analyses using BLASTN showed that pEI70 is almost entirely included in plasmid pEB102 from E. billingiae, an epiphytic Erwinia of pome fruits, with sequence identities superior to 98%. A duplex PCR assay was developed to survey the prevalence of plasmid pEI70 and also that of pEA29, which had previously been described in several E. amylovora strains. Plasmid pEI70 was found widely dispersed across Europe with frequencies of 5-92%, but it was absent in E. amylovora analyzed populations from outside of Europe. Restriction analysis and hybridization demonstrated that this plasmid was identical in at least 13 strains. Curing E. amylovora strains of pEI70 reduced their aggressiveness on pear, and introducing pEI70 into low-aggressiveness strains lacking this plasmid increased symptoms development in this host. Discovery of this novel plasmid offers new insights into the biogeography, evolution and virulence determinants in E. amylovora.

  2. Pathogenicity and infection strategies of the fire blight pathogen Erwinia amylovora in Rosaceae: state of the art.

    Science.gov (United States)

    Vrancken, K; Holtappels, M; Schoofs, H; Deckers, T; Valcke, R

    2013-05-01

    Plants are host to a large amount of pathogenic bacteria. Fire blight, caused by the bacterium Erwinia amylovora, is an important disease in Rosaceae. Pathogenicity of E. amylovora is greatly influenced by the production of exopolysaccharides, such as amylovoran, and the use of the type III secretion system, which enables bacteria to penetrate host tissue and cause disease. When infection takes place, plants have to rely on the ability of each cell to recognize the pathogen and the signals emanating from the infection site in order to generate several defence mechanisms. These mechanisms consist of physical barriers and the production of antimicrobial components, both in a preformed and an inducible manner. Inducible defence responses are activated upon the recognition of elicitor molecules by plant cell receptors, either derived from invading micro-organisms or from pathogen-induced degradation of plant tissue. This recognition event triggers a signal transduction cascade, leading to a range of defence responses [reactive oxygen species (ROS), plant hormones, secondary metabolites, …] and redeployment of cellular energy in a fast, efficient and multiresponsive manner, which prevents further pathogen ingress. This review highlights the research that has been performed during recent years regarding this specific plant-pathogen interaction between Erwinia amylovora and Rosaceae, with a special emphasis on the pathogenicity and the infection strategy of E. amylovora and the possible defence mechanisms of the plant against this disease.

  3. The Differences among Pear Genotypes to Fire Blight (Erwinia amylovora Attack, Based on Observations of Natural Infection

    Directory of Open Access Journals (Sweden)

    Adriana F. SESTRAS

    2008-08-01

    Full Text Available Fire blight, caused by the bacterium Erwinia amylovora, is one of the most damaging diseases of pear in the world. In Cluj-Napoca area, situated in central Transylvania, Romania, fire blight was observed first in 1994, very late comparative with the other countries from occidental Europe. The response of the pear cultivars and species from National Pear Collection from Cluj-Napoca to fire blight attack, assessed in natural conditions of infection, range on a large scale of variability, which denotes a strong influence of the genotype in expression of resistance or sensitivity to disease. From all genotypes, about 20.5% have not presented symptoms of attack, among them being the following: 'Blanquet precoce', 'Klementinka', 'Severianka', 'Beurre Bachelier', 'Kieffer Seedling', 'Er Shi Shinge', 'Beurre Amanlis', 'Bristol Cross', 'Beurre Liegel', 'Beurre Lucon', 'Grand Champion', 'Magness', 'Mericourt' etc. and several ancient autochthonous cultivars ('Pere malaiete', 'De zahar de Bihor', 'Cu miez rosu', 'Clopotele', 'Garoafa mare', 'Craiese', 'Para de apa'. Also, there were identified several species of Pyrus with no attack, as P. pollveria, P. common pear, P. lindlezi, P. malifolia, P. persica, P. ussuriensis, P. variolosa. The remarked genotypes could be potential sources for further breeding programmes and increase the number of genotypes available for breeding new pear cultivars resistant to Erwinia attack.

  4. Virulence Factors of Erwinia amylovora: A Review.

    Science.gov (United States)

    Piqué, Núria; Miñana-Galbis, David; Merino, Susana; Tomás, Juan M

    2015-06-05

    Erwinia amylovora, a Gram negative bacteria of the Enterobacteriaceae family, is the causal agent of fire blight, a devastating plant disease affecting a wide range of host species within Rosaceae and a major global threat to commercial apple and pear production. Among the limited number of control options currently available, prophylactic application of antibiotics during the bloom period appears the most effective. Pathogen cells enter plants through the nectarthodes of flowers and other natural openings, such as wounds, and are capable of rapid movement within plants and the establishment of systemic infections. Many virulence determinants of E. amylovora have been characterized, including the Type III secretion system (T3SS), the exopolysaccharide (EPS) amylovoran, biofilm formation, and motility. To successfully establish an infection, E. amylovora uses a complex regulatory network to sense the relevant environmental signals and coordinate the expression of early and late stage virulence factors involving two component signal transduction systems, bis-(3'-5')-cyclic di-GMP (c-di-GMP) and quorum sensing. The LPS biosynthetic gene cluster is one of the relatively few genetic differences observed between Rubus- and Spiraeoideae-infecting genotypes of E. amylovora. Other differential factors, such as the presence and composition of an integrative conjugative element associated with the Hrp T3SS (hrp genes encoding the T3SS apparatus), have been recently described. In the present review, we present the recent findings on virulence factors research, focusing on their role in bacterial pathogenesis and indicating other virulence factors that deserve future research to characterize them.

  5. Microbiological examination of Erwinia amylovora exopolysaccharide ooze.

    Science.gov (United States)

    Slack, Suzanne; Zeng, Quan; Outwater, Cory; Sundin, George W

    2017-01-03

    Fire blight, caused by the pathogen Erwinia amylovora (Burrill) Winslow et al, is the most devastating bacterial disease of pome fruits in North America and worldwide. The primary method of dispersal for E. amylovora is through ooze, a mass of exopolysaccharides and bacterial cells that is exuded as droplets from infected host tissue. During the 2013 and 2014 field seasons, 317 ooze droplets were collected from field-inoculated apple trees. Populations of E. amylovora in ooze droplets were 108 colony forming units per microliter on average (cfu/µl). Ooze droplets harboring larger (> 108 cfu/µl) cell populations were typically smaller in total volume and had darker coloring, such as orange, red, or dark red hues. Examination of apple host tissue at the site of emergence of ooze droplets using scanning electron microscopy revealed that ooze was not exuding through natural openings; instead, it was found on erumpent mounds and small (10 µm) tears in tissue. These observations suggested that E. amylovora-induced wounds in tissue provided the exit holes for ooze extrusion from the host. Analyses of E. amylovora populations in ooze droplets and within the stems from which ooze droplets emerged indicated that approximately 9% of the total bacterial population from infected stems is diverted to ooze. Gene expression analyses indicated that E. amylovora cells in stem sections located above ooze droplets and in ooze droplets were actively expressing critical pathogenicity genes such as hrpL, dspE, and amsK. Thus, our study identified ooze as a source of large, concentrated populations of E. amylovora that emerged from the host by rupturing host tissue. As the cells in ooze droplets are expressing genes required for pathogenesis, they are already primed for infection should they be dispersed from ooze to new infection courts.

  6. Virulence Factors of Erwinia amylovora: A Review

    Directory of Open Access Journals (Sweden)

    Núria Piqué

    2015-06-01

    Full Text Available Erwinia amylovora, a Gram negative bacteria of the Enterobacteriaceae family, is the causal agent of fire blight, a devastating plant disease affecting a wide range of host species within Rosaceae and a major global threat to commercial apple and pear production. Among the limited number of control options currently available, prophylactic application of antibiotics during the bloom period appears the most effective. Pathogen cells enter plants through the nectarthodes of flowers and other natural openings, such as wounds, and are capable of rapid movement within plants and the establishment of systemic infections. Many virulence determinants of E. amylovora have been characterized, including the Type III secretion system (T3SS, the exopolysaccharide (EPS amylovoran, biofilm formation, and motility. To successfully establish an infection, E. amylovora uses a complex regulatory network to sense the relevant environmental signals and coordinate the expression of early and late stage virulence factors involving two component signal transduction systems, bis-(3′-5′-cyclic di-GMP (c-di-GMP and quorum sensing. The LPS biosynthetic gene cluster is one of the relatively few genetic differences observed between Rubus- and Spiraeoideae-infecting genotypes of E. amylovora. Other differential factors, such as the presence and composition of an integrative conjugative element associated with the Hrp T3SS (hrp genes encoding the T3SS apparatus, have been recently described. In the present review, we present the recent findings on virulence factors research, focusing on their role in bacterial pathogenesis and indicating other virulence factors that deserve future research to characterize them.

  7. Cloning, expression, purification, crystallization and preliminary X-ray analysis of EaLsc, a levansucrase from Erwinia amylovora.

    Science.gov (United States)

    Caputi, Lorenzo; Cianci, Michele; Benini, Stefano

    2013-05-01

    The Gram-negative bacterium Erwinia amylovora is a destructive pathogen of Rosaceae. During infection, E. amylovora produces the exopolysaccharide levan, which contributes to the occlusion of plant vessels, causing the wilting of shoots. Levan is a fructose polymer that is synthesized by multifunctional enzymes called levansucrases. The levansucrase from E. amylovora (EaLsc) was heterologously expressed as a GST-fusion protein in Escherichia coli, purified and crystallized after tag removal. The protein crystallized in space group P21212. X-ray diffraction data were acquired to 2.77 Å resolution. The structure of the enzyme was solved by molecular replacement. The asymmetric unit contains eight enzyme molecules, giving a solvent content of 58.74% and a Matthews coefficient of 2.98 Å(3) Da(-1).

  8. Growth inhibition of Erwinia amylovora and related Erwinia species by neutralized short‑chain fatty acids.

    Science.gov (United States)

    Konecki, Katrin; Gernold, Marina; Wensing, Annette; Geider, Klaus

    2013-11-01

    Short-chain fatty acids (SCFAs) are used to preserve food and could be a tool for control of fire blight caused by Erwinia amylovora on apple, pear and related rosaceous plants. Neutralized acids were added to buffered growth media at 0.5–75 mM and tested at pHs ranging from 6.8 to 5.5. Particularly at low pH, SCFAs with a chain length exceeding that of acetic acid such as propionic acid were effective growth inhibitors of E. amylovora possibly due to uptake of free acid and its intracellular accumulation. We also observed high inhibition with monochloroacetic acid. An E. billingiae strain was as sensitive to the acids as E. amylovora or E. tasmaniensis. Fire blight symptoms on pear slices were reduced when the slices were pretreated with neutralized propionic acid. Propionic acid is well water soluble and could be applied in orchards as a control agent for fire blight.

  9. Erwinia tracheiphila colonization of cantaloupe fruits through flower inoculation

    Science.gov (United States)

    Cantaloupe (Cucumis melo var. cantalupensis) is a nutritious fresh fruit. Bacterial wilt, caused by Erwinia tracheiphila, is the most devastating cantaloupe disease globally. The pathogen is transmitted in nature by xylem-feeding spotted and striped cucumber beetles; other modes of infection have ...

  10. Hyacintentelers stemmen in met aanpak Erwinia en fonds onderzoek

    NARCIS (Netherlands)

    Dwarswaard, A.; Vreeburg, P.J.M.

    2013-01-01

    Twee stevige onderwerpen telde de agenda van de jaarvergadering van de KAVB-productgroep Hyacint. Het voorstel om werkbollen te gaan toetsen op Erwinia kreeg bijval, en ook het plan om een fonds te vormen waaruit hyacintenonderzoek wordt betaald werd met instemming begroet.

  11. Fysische, chemische en biologische bestrijding van pectinolytische Erwinia's

    NARCIS (Netherlands)

    Wolf, van der J.M.; Doorn, van J.

    2006-01-01

    Rotveroorzakende bacteriën, met name pectinolytische Erwinia spp., zijn verantwoordelijk voor veel schade in de diverse gewassen, vooral in de teelt van aardappelen en de bloembolgewassen hyacint, zantedeschia en iris. In deze literatuurstudie worden de beschikbare gegevens nog eens nader bekeken va

  12. Proteins from Erwinia asparaginase Erwinase ® and E. coli asparaginase 2 MEDAC ® for treatment of human leukemia, show a multitude of modifications for which the consequences are completely unclear.

    Science.gov (United States)

    Bae, Narkhyun; Pollak, Arnold; Lubec, Gert

    2011-07-01

    L-Asparaginase from Erwinia chrysanthemi (ASPG_ERWCH; UniProtKB accession number P06608 (Erwinase(®))) and L-asparaginase 2 from Escherichia coli (ASPG2_ECOLI; UniProtKB accession number P00805 (Medac(®))), both L-asparagine amidohydrolases, are widely used for the treatment of acute lymphoblastic leukemia. A series of serious side effects have been reported and this warrants studies into the protein chemistry of the medical products sold. Mass spectrometry (MS) data on ASPG_ERWCH and ASPG2_ECOLI have not been published so far and herein a gel-based proteomics study was performed to provide information about sequence and modifications of the commercially available medical products. ASPG_ERWCH and ASPG2_ECOLI were applied onto two-dimensional gel electrophoresis, spots were in-gel digested with several proteases and resulting peptides and protein modifications were analysed by nano-ESI-LC-MS/MS. Four spots were observed for ASPG_ERWCH, six spots were observed for ASPG2_ECOLI and the identified proteins showed high sequence coverage without sequence conflicts. Several protein modifications including technical and posttranslational modifications were demonstrated. Protein modifications are known to change physicochemical, immunochemical, biological and pharmacological properties and results from this work may challenge re-designing of the product including possible removal of the modifications by the manufacturer because it is not known whether they are contributing to the serious adverse effects of the protein drug.

  13. Cellular, physiological, and molecular adaptive responses of Erwinia amylovora to starvation.

    Science.gov (United States)

    Santander, Ricardo D; Oliver, James D; Biosca, Elena G

    2014-05-01

    Erwinia amylovora causes fire blight, a destructive disease of rosaceous plants distributed worldwide. This bacterium is a nonobligate pathogen able to survive outside the host under starvation conditions, allowing its spread by various means such as rainwater. We studied E. amylovora responses to starvation using water microcosms to mimic natural oligotrophy. Initially, survivability under optimal (28 °C) and suboptimal (20 °C) growth temperatures was compared. Starvation induced a loss of culturability much more pronounced at 28 °C than at 20 °C. Natural water microcosms at 20 °C were then used to characterize cellular, physiological, and molecular starvation responses of E. amylovora. Challenged cells developed starvation-survival and viable but nonculturable responses, reduced their size, acquired rounded shapes and developed surface vesicles. Starved cells lost motility in a few days, but a fraction retained flagella. The expression of genes related to starvation, oxidative stress, motility, pathogenicity, and virulence was detected during the entire experimental period with different regulation patterns observed during the first 24 h. Further, starved cells remained as virulent as nonstressed cells. Overall, these results provide new knowledge on the biology of E. amylovora under conditions prevailing in nature, which could contribute to a better understanding of the life cycle of this pathogen.

  14. Phylogeography and population structure of the biologically invasive phytopathogen Erwinia amylovora inferred using minisatellites.

    Science.gov (United States)

    Bühlmann, Andreas; Dreo, Tanja; Rezzonico, Fabio; Pothier, Joël F; Smits, Theo H M; Ravnikar, Maja; Frey, Jürg E; Duffy, Brion

    2014-07-01

    Erwinia amylovora causes a major disease of pome fruit trees worldwide, and is regulated as a quarantine organism in many countries. While some diversity of isolates has been observed, molecular epidemiology of this bacterium is hindered by a lack of simple molecular typing techniques with sufficiently high resolution. We report a molecular typing system of E. amylovora based on variable number of tandem repeats (VNTR) analysis. Repeats in the E. amylovora genome were identified with comparative genomic tools, and VNTR markers were developed and validated. A Multiple-Locus VNTR Analysis (MLVA) was applied to E. amylovora isolates from bacterial collections representing global and regional distribution of the pathogen. Based on six repeats, MLVA allowed the distinction of 227 haplotypes among a collection of 833 isolates of worldwide origin. Three geographically separated groups were recognized among global isolates using Bayesian clustering methods. Analysis of regional outbreaks confirmed presence of diverse haplotypes but also high representation of certain haplotypes during outbreaks. MLVA analysis is a practical method for epidemiological studies of E. amylovora, identifying previously unresolved population structure within outbreaks. Knowledge of such structure can increase our understanding on how plant diseases emerge and spread over a given geographical region.

  15. Exopolysaccharides favor the survival of Erwinia amylovora under copper stress through different strategies.

    Science.gov (United States)

    Ordax, Mónica; Marco-Noales, Ester; López, María M; Biosca, Elena G

    2010-09-01

    Erwinia amylovora causes fire blight, a destructive disease of rosaceous plants very difficult to control. We demonstrated that copper, employed to control plant diseases, induces the "viable-but-nonculturable" (VBNC) state in E. amylovora. Moreover, it was previously reported that copper increases production of its main exopolysaccharide (EPS), amylovoran. In this work, the copper-complexing ability of amylovoran and levan, other major EPS of E. amylovora, was demonstrated. Following this, EPS-deficient mutants were used to determine the role of these EPSs in survival of this bacterium in AB mineral medium with copper, compared to their wild type strain and AB without copper. Total, viable and culturable counts of all strains were monitored for six months. With copper, a larger fraction of the viable population of EPS mutants entered into the VBNC state, and earlier than their wild type strain, showing the contribution of both EPSs to long-term survival in a culturable state. Further, we demonstrated that both EPSs can be used as carbon source by E. amylovora under deprivation conditions. Overall, these previously unreported functions of amylovoran and levan provide survival advantages for E. amylovora, which could contribute to its enhanced persistence in nature.

  16. Medfly Ceratitis capitata as Potential Vector for Fire Blight Pathogen Erwinia amylovora: Survival and Transmission.

    Directory of Open Access Journals (Sweden)

    Mónica Ordax

    Full Text Available Monitoring the ability of bacterial plant pathogens to survive in insects is required for elucidating unknown aspects of their epidemiology and for designing appropriate control strategies. Erwinia amylovora is a plant pathogenic bacterium that causes fire blight, a devastating disease in apple and pear commercial orchards. Studies on fire blight spread by insects have mainly focused on pollinating agents, such as honeybees. However, the Mediterranean fruit fly (medfly Ceratitis capitata (Diptera: Tephritidae, one of the most damaging fruit pests worldwide, is also common in pome fruit orchards. The main objective of the study was to investigate whether E. amylovora can survive and be transmitted by the medfly. Our experimental results show: i E. amylovora can survive for at least 8 days inside the digestive tract of the medfly and until 28 days on its external surface, and ii medflies are able to transmit the bacteria from inoculated apples to both detached shoots and pear plants, being the pathogen recovered from lesions in both cases. This is the first report on E. amylovora internalization and survival in/on C. capitata, as well as the experimental transmission of the fire blight pathogen by this insect. Our results suggest that medfly can act as a potential vector for E. amylovora, and expand our knowledge on the possible role of these and other insects in its life cycle.

  17. Identification and characterization of Nip, necrosis-inducing virulence protein of Erwinia carotovora subsp. carotovora.

    Science.gov (United States)

    Mattinen, Laura; Tshuikina, Marina; Mäe, Andres; Pirhonen, Minna

    2004-12-01

    Erwinia carotovora subsp. carotovora is a gram-negative bacterium that causes soft rot disease of many cultivated crops. When a collection of E. carotovora subsp. carotovora isolates was analyzed on a Southern blot using the harpin-encoding gene hrpN as probe, several harpinless isolates were found. Regulation of virulence determinants in one of these, strain SCC3193, has been characterized extensively. It is fully virulent on potato and in Arabidopsis thaliana. An RpoS (SigmaS) mutant of SCC3193, producing elevated levels of secreted proteins, was found to cause lesions resembling the hypersensitive response when infiltrated into tobacco leaf tissue. This phenotype was evident only when bacterial cells had been cultivated on solid minimal medium at low pH and temperature. The protein causing'the cell death was purified and sequenced, and the corresponding gene was cloned. The deduced sequence of the necrosis-inducing protein (Nip) showed homology to necrosis- and ethylene-inducing elicitors of fungi and oomycetes. A mutant strain of E. carotovora subsp. carotovora lacking the nip gene showed reduced virulence in potato tuber assay but was unaffected in virulence in potato stem or on other tested host plants.

  18. Medfly Ceratitis capitata as Potential Vector for Fire Blight Pathogen Erwinia amylovora: Survival and Transmission.

    Science.gov (United States)

    Ordax, Mónica; Piquer-Salcedo, Jaime E; Santander, Ricardo D; Sabater-Muñoz, Beatriz; Biosca, Elena G; López, María M; Marco-Noales, Ester

    2015-01-01

    Monitoring the ability of bacterial plant pathogens to survive in insects is required for elucidating unknown aspects of their epidemiology and for designing appropriate control strategies. Erwinia amylovora is a plant pathogenic bacterium that causes fire blight, a devastating disease in apple and pear commercial orchards. Studies on fire blight spread by insects have mainly focused on pollinating agents, such as honeybees. However, the Mediterranean fruit fly (medfly) Ceratitis capitata (Diptera: Tephritidae), one of the most damaging fruit pests worldwide, is also common in pome fruit orchards. The main objective of the study was to investigate whether E. amylovora can survive and be transmitted by the medfly. Our experimental results show: i) E. amylovora can survive for at least 8 days inside the digestive tract of the medfly and until 28 days on its external surface, and ii) medflies are able to transmit the bacteria from inoculated apples to both detached shoots and pear plants, being the pathogen recovered from lesions in both cases. This is the first report on E. amylovora internalization and survival in/on C. capitata, as well as the experimental transmission of the fire blight pathogen by this insect. Our results suggest that medfly can act as a potential vector for E. amylovora, and expand our knowledge on the possible role of these and other insects in its life cycle.

  19. Molecular Weight and Aggregation of Erwinia Gum in Aqueous Solutions

    Institute of Scientific and Technical Information of China (English)

    2000-01-01

    Erwinia(E) gum is composed of glucose, fucose, galactose and glucuronic acid. The weight-average molecular weights Mw, number-average molecular weights Mn and intrinsic viscosities[η] of the four fractions and the unfractionated E gum in aqueous solutions at desired temperatures were studied by light scattering, membrane osmometry, size exclusion chromatography(SEC) and viscometry. The experimental results prove that E gum formed aggregates in the aqueous solution at 25 ℃ and the aggregates were broken gradually with increasing temperature. The dissociation of the aggregates of E gum in the aqueous solution started at 36 ℃, and was completed at around 90 ℃. The [η] values of E gum and its fractions are much higher than those of the conventional polymers with the similar molecular weights, and decrease with increasing NaCl concentration.

  20. Characterization of the RcsC sensor kinase from Erwinia amylovora and other Enterobacteria.

    Science.gov (United States)

    Wang, Dongping; Korban, Schuyler S; Pusey, P Lawrence; Zhao, Youfu

    2011-06-01

    RcsC is a hybrid sensor kinase which contains a sensor domain, a histidine kinase domain, and a receiver domain. We have previously demonstrated that, although the Erwinia amylovora rcsC mutant produces more amylovoran than the wild-type (WT) strain in vitro, the mutant remains nonpathogenic on both immature pear fruit and apple plants. In this study, we have comparatively characterized the Erwinia RcsC and its homologs from various enterobacteria. Results demonstrate that expression of the Erwinia rcsC gene suppresses amylovoran production in various amylovoran overproducing WT and mutant strains, thus suggesting the presence of a net phosphatase activity of Erwinia RcsC. Findings have also demonstrated that rcsC homologs from other enterobacteria could not rescue amylovoran production of the Erwinia rcsC mutant in vitro. However, virulence of the Erwinia rcsC mutant is partially restored by rcsC homologs from Pantoea stewartii, Yersinia pestis, and Salmonella enterica but not from Escherichia coli on apple shoots. Domain-swapping experiments have indicated that replacement of the E. coli RcsC sensor domain by those of Erwinia and Yersinia spp. partially restores virulence of the Erwinia rcsC mutant, whereas chimeric constructs containing the sensor domain of E. coli RcsC could not rescue virulence of the Erwinia rcsC mutant on apple. Interestingly, only chimeric constructs containing the histidine kinase and receiver domains of Erwinia RcsC are fully capable of rescuing amylovoran production. These results suggest that the sensor domain of RcsC may be important in regulating bacterial virulence, whereas the activity of the histidine kinase and receiver domains of Erwinia RcsC may be essential for amylovoran production in vitro.

  1. Efficacy of potato seeds disinfection products to control Erwinia spp.

    Science.gov (United States)

    Dupuis, B; Garcia, N; Boels, G

    2008-01-01

    Erwinia spp. provokes soft rot on potato tubers during storage. No disinfection products are available on the market in the European Union to control these bacteria. We tested 3 products presented as good candidates to cure potato tubers from bacterial diseases. First, Anthium 500 (Du Pont de Nemours) a product based on chlorine dioxyde, then Phostrol (Nufarm) with phosphoric acid as a.i. and finally Solucuivre (Proval), a copper based product. We firstly managed disinfection trials: high Erwinia contaminated potato seed samples were treated by immersion and were then incubated, we observed the percentage of tubers rotting. Secondly, we managed protection trials: protected healthy tubers were incubated during 23 days in contact with rotting tubers. We evaluated weight loss after symptoms development. No tested product was effective to control Erwinia spp. on seed tubers in our trials conditions. Furthermore, we observed more rot development after Phostrol and Solucuivre application. We suppose that the product couldn't reach the latent bacteria and weakened the tubers. No protection effect was observed.

  2. Draft Genome Sequence of the Bactrocera oleae Symbiont “Candidatus Erwinia dacicola”

    Science.gov (United States)

    Blow, Frances; Gioti, Anastasia; Starns, David; Ben-Yosef, Michael; Pasternak, Zohar; Jurkevitch, Edouard; Vontas, John

    2016-01-01

    “Candidatus Erwinia dacicola” is a Gammaproteobacterium that forms a symbiotic association with the agricultural pest Bactrocera oleae. Here, we present a 2.1-Mb draft hybrid genome assembly for “Ca. Erwinia dacicola” generated from single-cell and metagenomic data. PMID:27634990

  3. Goed om uitgangsmateriaal op Erwinia te testen (interview met Jan van der Wolf)

    NARCIS (Netherlands)

    Dwarswaard, A.; Bovenkamp, van den G.; Wolf, van der J.M.

    2012-01-01

    Bloembollen en aardappelen. Ze hebben in ieder geval één ziekte gemeen: bacterierot, veroorzaakt door Erwinia. In beide teelten zorgen deze bacterieziekten voor veel schade. In het Deltaplan Erwinia werken de bloembollen- en aardappelwereld samen op onderzoeksgebied. In twee artikelen staat de vraag

  4. Complete genome sequence of Japanese erwinia strain ejp617, a bacterial shoot blight pathogen of pear.

    Science.gov (United States)

    Park, Duck Hwan; Thapa, Shree Prasad; Choi, Beom-Soon; Kim, Won-Sik; Hur, Jang Hyun; Cho, Jun Mo; Lim, Jong-Sung; Choi, Ik-Young; Lim, Chun Keun

    2011-01-01

    The Japanese Erwinia strain Ejp617 is a plant pathogen that causes bacterial shoot blight of pear in Japan. Here, we report the complete genome sequence of strain Ejp617 isolated from Nashi pears in Japan to provide further valuable insight among related Erwinia species.

  5. Erwinia uzenensis sp. nov., a novel pathogen that affects European pear trees (Pyrus communis L.).

    Science.gov (United States)

    Matsuura, Takayuki; Mizuno, Akifumi; Tsukamoto, Takanori; Shimizu, Yoshiaki; Saito, Norihiko; Sato, Shigeyoshi; Kikuchi, Shigemi; Uzuki, Tsuneyasu; Azegami, Koji; Sawada, Hiroyuki

    2012-08-01

    Bacteria were isolated from black lesions on shoots of European pear trees (Pyrus communis L.) in an orchard in Japan. Previous characterization of this novel pathogen by phenotypic and genotypic methods suggested that it should belong to the genus Erwinia but might not correspond to either Erwinia amylovora or Erwinia pyrifoliae. Here, phylogenetic analyses of the 16S rRNA gene, gyrB, and rpoD gene sequences indicated that it could not be assigned to any recognized species of the genus Erwinia. DNA-DNA hybridization confirmed that the bacterial strains represented a novel species. The DNA G+C contents, the fatty acid profile and phenotypic characteristics resembled those previously reported for members of the genus Erwinia. On the basis of these and previous results, the pathogen represents a novel species of the genus Erwinia, for which the name Erwinia uzenensis sp. nov. (type strain: YPPS 951(T) = LMG 25843(T) = NCPPB 4475(T)) is proposed.

  6. Erwinia teleogrylli sp. nov., a Bacterial Isolate Associated with a Chinese Cricket.

    Directory of Open Access Journals (Sweden)

    Bo Liu

    Full Text Available A bacterial isolate (SCU-B244T was obtained in China from crickets (Teleogryllus occipitalis living in cropland deserted for approximately 10 years. The isolated bacteria were Gram-negative, facultatively anaerobic, oxidase-negative rods. A preliminary analysis of the 16S rRNA gene sequence indicated that the strain belongs to either the genus Erwinia or Pantoea. Analysis of multilocus sequence typing based on concatenated partial atpD, gyrB and infB gene sequences and physiological and biochemical characteristics indicated that the strain belonged to the genus Erwinia, as member of a new species as it was distinct from other known Erwinia species. Further analysis of the 16S rRNA gene showed SCU-B244T to have 94.71% identity to the closest species of that genus, Erwinia oleae (DSM 23398T, which is below the threshold of 97% used to discriminate bacterial species. DNA-DNA hybridization results (5.78±2.52% between SCU-B244T and Erwinia oleae (DSM 23398T confirmed that SCU-B244T and Erwinia oleae (DSM 23398T represent different species combined with average nucleotide identity values which range from 72.42% to 74.41. The DNA G+C content of SCU-B244T was 55.32 mol%, which also differs from that of Erwinia oleae (54.7 to 54.9 mol%. The polyphasic taxonomic approach used here confirmed that the strain belongs to the Erwinia group and represents a novel species. The name Erwinia teleogrylli sp. nov. is proposed for this novel taxon, for which the type strain is SCU-B244T (= CGMCC 1.12772T = DSM 28222T = KCTC 42022T.

  7. Erwinia teleogrylli sp. nov., a Bacterial Isolate Associated with a Chinese Cricket.

    Science.gov (United States)

    Liu, Bo; Luo, Jin; Li, Wei; Long, Xiu-Feng; Zhang, Yu-Qin; Zeng, Zhi-Gang; Tian, Yong-Qiang

    2016-01-01

    A bacterial isolate (SCU-B244T) was obtained in China from crickets (Teleogryllus occipitalis) living in cropland deserted for approximately 10 years. The isolated bacteria were Gram-negative, facultatively anaerobic, oxidase-negative rods. A preliminary analysis of the 16S rRNA gene sequence indicated that the strain belongs to either the genus Erwinia or Pantoea. Analysis of multilocus sequence typing based on concatenated partial atpD, gyrB and infB gene sequences and physiological and biochemical characteristics indicated that the strain belonged to the genus Erwinia, as member of a new species as it was distinct from other known Erwinia species. Further analysis of the 16S rRNA gene showed SCU-B244T to have 94.71% identity to the closest species of that genus, Erwinia oleae (DSM 23398T), which is below the threshold of 97% used to discriminate bacterial species. DNA-DNA hybridization results (5.78±2.52%) between SCU-B244T and Erwinia oleae (DSM 23398T) confirmed that SCU-B244T and Erwinia oleae (DSM 23398T) represent different species combined with average nucleotide identity values which range from 72.42% to 74.41. The DNA G+C content of SCU-B244T was 55.32 mol%, which also differs from that of Erwinia oleae (54.7 to 54.9 mol%). The polyphasic taxonomic approach used here confirmed that the strain belongs to the Erwinia group and represents a novel species. The name Erwinia teleogrylli sp. nov. is proposed for this novel taxon, for which the type strain is SCU-B244T (= CGMCC 1.12772T = DSM 28222T = KCTC 42022T).

  8. Erwinia amylovora type three-secreted proteins trigger cell death and defense responses in Arabidopsis thaliana.

    Science.gov (United States)

    Degrave, A; Fagard, M; Perino, C; Brisset, M N; Gaubert, S; Laroche, S; Patrit, O; Barny, M-A

    2008-08-01

    Erwinia amylovora is the bacterium responsible for fire blight, a necrotic disease affecting plants of the rosaceous family. E. amylovora pathogenicity requires a functional type three secretion system (T3SS). We show here that E. amylovora triggers a T3SS-dependent cell death on Arabidopsis thaliana. The plants respond by inducing T3SS-dependent defense responses, including salicylic acid (SA)-independent callose deposition, activation of the SA defense pathway, reactive oxygen species (ROS) accumulation, and part of the jasmonic acid/ethylene defense pathway. Several of these reactions are similar to what is observed in host plants. We show that the cell death triggered by E. amylovora on A. thaliana could not be simply explained by the recognition of AvrRpt2 ea by the resistance gene product RPS2. We then analyzed the role of type three-secreted proteins (T3SPs) DspA/E, HrpN, and HrpW in the induction of cell death and defense reactions in A. thaliana following infection with the corresponding E. amylovora mutant strains. HrpN and DspA/E were found to play an important role in the induction of cell death, activation of defense pathways, and ROS accumulation. None of the T3SPs tested played a major role in the induction of SA-independent callose deposition. The relative importance of T3SPs in A. thaliana is correlated with their relative importance in the disease process on host plants, indicating that A. thaliana can be used as a model to study their role.

  9. Differentiation of Erwinia amylovora and Erwinia pyrifoliae strains with single nucleotide polymorphisms and by synthesis of dihydrophenylalanine.

    Science.gov (United States)

    Gehring, I; Geider, K

    2012-07-01

    Fire blight has spread from North America to New Zealand, Europe, and the Mediterranean region. We were able to differentiate strains from various origins with a novel PCR method. Three Single Nucleotide Polymorphisms (SNPs) in the Erwinia amylovora genome were characteristic of isolates from North America and could distinguish them from isolates from other parts of the world. They were derived from the galE, acrB, and hrpA genes of strains Ea273 and Ea1/79. These genes were analyzed by conventional PCR (cPCR) and quantitative PCR (qPCR) with differential primer annealing temperatures. North-American E. amylovora strains were further differentiated according to their production of L: -2,5-dihydrophenylalanine (DHP) as tested by growth inhibition of the yeast Rhodotorula glutinis. E. amylovora fruit tree (Maloideae) and raspberry (rubus) strains were also differentiated by Single Strand Conformational Polymorphism analysis. Strains from the related species Erwinia pyrifoliae isolated in Korea and Japan were all DHP positive, but were differentiated from each other by SNPs in the galE gene. Differential PCR is a rapid and simple method to distinguish E. amylovora as well as E. pyrifoliae strains according to their geographical origin.

  10. A gene cluster for amylovoran synthesis in Erwinia amylovora: characterization and relationship to cps genes in Erwinia stewartii.

    Science.gov (United States)

    Bernhard, F; Coplin, D L; Geider, K

    1993-05-01

    A large ams gene cluster required for production of the acidic extracellular polysaccharide (EPS) amylovoran by the fire blight pathogen Erwinia amylovora was cloned. Tn5 mutagenesis and gene replacement were used to construct chromosomal ams mutants. Five complementation groups, essential for amylovoran synthesis and virulence in E. amylovora, were identified and designated ams A-E. The ams gene cluster is about 7 kb in size and functionally equivalent to the cps gene cluster involved in EPS synthesis by the related pathogen Erwinia stewartii. Mucoidy and virulence were restored to E. stewartii mutants in four cps complementation groups by the cloned E. amylovora ams genes. Conversely, the E. stewartii cps gene cluster was able to complement mutations in E. amylovora ams genes. Correspondence was found between the amsA-E complementation groups and the cpsB-D region, but the arrangement of the genes appears to be different. EPS production and virulence were also restored to E. amylovora amsE and E. stewartii cpsD mutants by clones containing the Rhizobium meliloti exo A gene.

  11. Regulatory genes and environmental regulation of amylovoran biosynthesis in Erwinia amylovora

    Science.gov (United States)

    The requirement of the exopolysaccharide amylovoran for Erwinia amylovora pathogenesis is well documented. However, regulation of amylovoran biosynthesis has not been comprehensively studied. We have previously reported that amylovoran production is strain-dependent in E. amylovora isolates. We have...

  12. An enrichment microsphere immunoassay for the detection of Pectobaterium atrosepticum and Dickeya dianthicola in potato tuber extracts

    NARCIS (Netherlands)

    Peters, J.; Sledz, V.; Bergervoet, J.H.W.; Wolf, van der J.M.

    2007-01-01

    An enrichment microsphere immunoassay (MIA) was developed, based on the Luminex xMAP® technology, for the simultaneous (duplex) detection of Pectobacterium atrosepticum (former name Erwinia carotovora subsp. atroseptica) (Pca) and Dickeya dianthicola (former name Erwinia chrysanthemi) (Dcd) in potat

  13. Erwinia iniecta sp. nov., isolated from Russian wheat aphid (Diuraphis noxia).

    Science.gov (United States)

    Campillo, Tony; Luna, Emily; Portier, Perrine; Fischer-Le Saux, Marion; Lapitan, Nora; Tisserat, Ned A; Leach, Jan E

    2015-10-01

    Short, Gram-negative-staining, rod-shaped bacteria were isolated from crushed bodies of Russian wheat aphid [Diuraphis noxia (Kurdjumov)] and artificial diets after Russian wheat aphid feeding. Based on multilocus sequence analysis involving the 16S rRNA, atpD, infB, gyrB and rpoB genes, these bacterial isolates constitute a novel clade in the genus Erwinia, and were most closely related to Erwinia toletana. Representative distinct strains within this clade were used for comparisons with related species of Erwinia. Phenotypic comparisons using four distinct strains and average nucleotide identity (ANI) measurements using two distinct draft genomes revealed that these strains form a novel species within the genus Erwinia. The name Erwinia iniecta sp. nov. is proposed, and strain B120T ( = CFBP 8182T = NCCB 100485T) was designated the type strain. Erwinia iniecta sp. nov. was not pathogenic to plants. However, virulence to the Russian wheat aphid was observed.

  14. Examining phylogenetic relationships of Erwinia and Pantoea species using whole genome sequence data.

    Science.gov (United States)

    Zhang, Yucheng; Qiu, Sai

    2015-11-01

    The genera Erwinia and Pantoea contain species that are devastating plant pathogens, non-pathogen epiphytes, and opportunistic human pathogens. However, some controversies persist in the taxonomic classification of these two closely related genera. The phylogenomic analysis of these two genera was investigated via a comprehensive analysis of 25 Erwinia genomes and 23 Pantoea genomes. Single-copy orthologs could be extracted from the Erwinia/Pantoea core-genome to reconstruct the Erwinia/Pantoea phylogeny. This tree has strong bootstrap support for almost all branches. We also estimated the in silico DNA-DNA hybridization (isDDH) and the average nucleotide identity (ANI) values between each genome; strains from the same species showed ANI values ≥96% and isDDH values >70%. These data confirm that whole genome sequence data provides a powerful tool to resolve the complex taxonomic questions of Erwinia/Pantoea, e.g. Pantoea agglomerans 299R was not clustered into a single group with other P. agglomerans strains, and the ANI values and isDDH values between them were Erwinia/Pantoea phylogeny.

  15. Erwinia oleae sp. nov., isolated from olive knots caused by Pseudomonas savastanoi pv. savastanoi.

    Science.gov (United States)

    Moretti, Chiaraluce; Hosni, Taha; Vandemeulebroecke, Katrien; Brady, Carrie; De Vos, Paul; Buonaurio, Roberto; Cleenwerck, Ilse

    2011-11-01

    Three endophytic bacterial isolates were obtained in Italy from olive knots caused by Pseudomonas savastanoi pv. savastanoi. Phenotypic tests in combination with 16S rRNA gene sequence analysis indicated a phylogenetic position for these isolates in the genera Erwinia or Pantoea, and revealed two other strains with highly similar 16S rRNA gene sequences (>99 %), CECT 5262 and CECT 5264, obtained in Spain from olive knots. Rep-PCR DNA fingerprinting of the five strains from olive knots with BOX, ERIC and REP primers revealed three groups of profiles that were highly similar to each other. Multilocus sequence analysis (MLSA) based on concatenated partial atpD, gyrB, infB and rpoB gene sequences indicated that the strains constituted a single novel species in the genus Erwinia. The strains showed general phenotypic characteristics typical of the genus Erwinia and whole genome DNA-DNA hybridization data confirmed that they represented a single novel species of the genus Erwinia. The strains showed DNA G+C contents ranging from 54.7 to 54.9 mol%. They could be discriminated from phylogenetically related species of the genus Erwinia by their ability to utilize potassium gluconate, l-rhamnose and d-arabitol, but not glycerol, inositol or d-sorbitol. The name Erwinia oleae sp. nov. (type strain DAPP-PG 531(T)= LMG 25322(T) = DSM 23398(T)) is proposed for this novel taxon.

  16. Autoinducer-2 of the fire blight pathogen Erwinia amylovora and other plant-associated bacteria.

    Science.gov (United States)

    Mohammadi, Mojtaba; Geider, Klaus

    2007-01-01

    Autoinducers are important for cellular communication of bacteria. The luxS gene has a central role in the synthesis of autoinducer-2 (AI-2). The gene was identified in a shotgun library of Erwinia amylovora and primers designed for PCR amplification from bacterial DNA. Supernatants of several Erwinia amylovora strains were assayed for AI-2 activity with a Vibrio harveyi mutant and were positive. Many other plant-associated bacteria also showed AI-2 activity such as Erwinia pyrifoliae and Erwinia tasmaniensis. The luxS genes of several bacteria were cloned, sequenced, and complemented Escherichia coli strain DH5alpha and a Salmonella typhimurium mutant, both defective in luxS, for synthesis of AI-2. Assays to detect AI-2 activity in culture supernatants of several Pseudomonas syringae pathovars failed, which may indicate the absence of AI-2 or synthesis of another type. Several reporter strains did not detect synthesis of an acyl homoserine lactone (AHL, AI-1) by Erwinia amylovora, but confirmed AHL-synthesis for Erwinia carotovora ssp. atroseptica and Pantoea stewartii.

  17. Effect of a waaL mutation on lipopolysaccharide composition, oxidative stress survival, and virulence in Erwinia amylovora.

    Science.gov (United States)

    Berry, Matthew C; McGhee, Gayle C; Zhao, Youfu; Sundin, George W

    2009-02-01

    Erwinia amylovora, the causal agent of fire blight, is an enterobacterial pathogen of Rosaceous plants including apple and pear. We have been studying the response of E. amylovora to oxidative stress because, during infection, the bacterium elicits an oxidative burst response in host plants. During the screening of a transposon mutant library for hydrogen peroxide sensitivity, we identified a mutant carrying an insertion in waaL, a gene involved in lipopolysaccharide biosynthesis, that was more sensitive to hydrogen peroxide than the parental wild-type strain. We also confirmed that a waaL mutant of Pseudomonas aeruginosa exhibited an increased sensitivity to hydrogen peroxide compared with the wild-type strain. The E. amylovora waaL mutant was also reduced in virulence, showed a decrease in twitching motility, and was more sensitive to polymyxin B than the wild type. Each of these phenotypes was complemented by the cloned waaL gene. Our results highlight the importance of the lipopolysaccharide layer to virulence in E. amylovora and the unexpected finding of an additional function of lipopolysaccharide in protection from oxidative stress in E. amylovora and P. aeruginosa.

  18. Role of electron transport chain of chloroplasts in oxidative burst of interaction between Erwinia amylovora and host cells.

    Science.gov (United States)

    Abdollahi, Hamid; Ghahremani, Zahra; Erfaninia, Kobra; Mehrabi, Rahim

    2015-05-01

    Erwinia amylovora is a necrogenic bacterium, causing the fire blight disease on many rosaceous plants. Triggering oxidative burst by E. amylovora is a key response by which host plants try to restrain pathogen spread. Electron transport chain (ETC) of chloroplasts is known as an inducible source of reactive oxygen species generation in various stresses. This research was performed to assess the role of this ETC in E. amylovora-host interaction using several inhibitors of this chain in susceptible and resistant apple and pear genotypes. All ETC inhibitors delayed appearance of disease necrosis, but the effects of methyl viologen, glutaraldehyde, and DCMU were more significant. In the absence of inhibitors, resistant genotypes showed an earlier and severe H2O2 generation and early suppression of redox dependent, psbA gene. The effects of inhibitors were corresponding to the redox potential of ETC inhibitory sites. In addition, delayed necrosis appearance was associated with the decreased disease severity and delayed H2O2 generation. These results provide evidences for the involvement of this ETC in host oxidative burst and suggest that chloroplast ETC has significant role in E. amylovora-host interaction.

  19. Eop1 from a Rubus strain of Erwinia amylovora functions as a host-range limiting factor.

    Science.gov (United States)

    Asselin, J E; Bonasera, J M; Kim, J F; Oh, C-S; Beer, S V

    2011-08-01

    Strains of Erwinia amylovora, the bacterium causing the disease fire blight of rosaceous plants, are separated into two groups based on host range: Spiraeoideae and Rubus strains. Spiraeoideae strains have wide host ranges, infecting plants in many rosaceous genera, including apple and pear. In the field, Rubus strains infect the genus Rubus exclusively, which includes raspberry and blackberry. Based on comparisons of limited sequence data from a Rubus and a Spiraeoideae strain, the gene eop1 was identified as unusually divergent, and it was selected as a possible host specificity factor. To test this, eop1 genes from a Rubus strain and a Spiraeoideae strain were cloned and mutated. Expression of the Rubus-strain eop1 reduced the virulence of E. amylovora in immature pear fruit and in apple shoots. Sequencing the orfA-eop1 regions of several strains of E. amylovora confirmed that forms of eop1 are conserved among strains with similar host ranges. This work provides evidence that eop1 from a Rubus-specific strain can function as a determinant of host specificity in E. amylovora.

  20. Control of plant defense mechanisms and fire blight pathogenesis through the regulation of 6-thioguanine biosynthesis in Erwinia amylovora.

    Science.gov (United States)

    Coyne, Sébastien; Litomska, Agnieszka; Chizzali, Cornelia; Khalil, Mohammed N A; Richter, Klaus; Beerhues, Ludger; Hertweck, Christian

    2014-02-10

    Fire blight is a devastating disease of Rosaceae plants, such as apple and pear trees. It is characterized by necrosis of plant tissue, caused by the phytopathogenic bacterium Erwinia amylovora. The plant pathogen produces the well-known antimetabolite 6-thioguanine (6TG), which plays a key role in fire blight pathogenesis. Here we report that YcfR, a member of the LTTR family, is a major regulator of 6TG biosynthesis in E. amylovora. Inactivation of the regulator gene (ycfR) led to dramatically decreased 6TG production. Infection assays with apple plants (Malus domestica cultivar Holsteiner Cox) and cell cultures of Sorbus aucuparia (mountain ash, rowan) revealed abortive fire blight pathogenesis and reduced plant response (biphenyl and dibenzofuran phytoalexin production). In the presence of the ΔycfR mutant, apple trees were capable of activating the abscission machinery to remove infected tissue. In addition to unveiling the regulation of 6TG biosynthesis in a major plant pathogen, we demonstrate for the first time that this antimetabolite plays a pivotal role in dysregulating the plant response to infection.

  1. Transcriptomic responses of the olive fruit fly Bactrocera oleae and its symbiont Candidatus Erwinia dacicola to olive feeding

    Science.gov (United States)

    Pavlidi, Nena; Gioti, Anastasia; Wybouw, Nicky; Dermauw, Wannes; Ben-Yosef, Michael; Yuval, Boaz; Jurkevich, Edouard; Kampouraki, Anastasia; Van Leeuwen, Thomas; Vontas, John

    2017-01-01

    The olive fruit fly, Bactrocera oleae, is the most destructive pest of olive orchards worldwide. The monophagous larva has the unique capability of feeding on olive mesocarp, coping with high levels of phenolic compounds and utilizing non-hydrolyzed proteins present, particularly in the unripe, green olives. On the molecular level, the interaction between B. oleae and olives has not been investigated as yet. Nevertheless, it has been associated with the gut obligate symbiotic bacterium Candidatus Erwinia dacicola. Here, we used a B.oleae microarray to analyze the gene expression of larvae during their development in artificial diet, unripe (green) and ripe (black) olives. The expression profiles of Ca. E. dacicola were analyzed in parallel, using the Illumina platform. Several genes were found overexpressed in the olive fly larvae when feeding in green olives. Among these, a number of genes encoding detoxification and digestive enzymes, indicating a potential association with the ability of B. oleae to cope with green olives. In addition, a number of biological processes seem to be activated in Ca. E. dacicola during the development of larvae in olives, with the most notable being the activation of amino-acid metabolism. PMID:28225009

  2. The phytoalexin-inducible multidrug efflux pump AcrAB contributes to virulence in the fire blight pathogen, Erwinia amylovora.

    Science.gov (United States)

    Burse, Antje; Weingart, Helge; Ullrich, Matthias S

    2004-01-01

    The enterobacterium Erwinia amylovora causes fire blight on members of the family Rosaceae, with economic importance on apple and pear. During pathogenesis, the bacterium is exposed to a variety of plant-borne antimicrobial compounds. In plants of Rosaceae, many constitutively synthesized isoflavonoids affecting microorganisms were identified. Bacterial multidrug efflux transporters which mediate resistance toward structurally unrelated compounds might confer tolerance to these phytoalexins. To prove this hypothesis, we cloned the acrAB locus from E. amylovora encoding a resistance nodulation division-type transport system. In Escherichia coli, AcrAB of E. amylovora conferred resistance to hydrophobic and amphiphilic toxins. An acrB-deficient E. amylovora mutant was impaired in virulence on apple rootstock MM 106. Furthermore, it was susceptible toward extracts of leaves of MM 106 as well as to the apple phytoalexins phloretin, naringenin, quercetin, and (+)-catechin. The expression of acrAB was determined using the promoterless reporter gene egfp. The acrAB operon was up-regulated in vitro by the addition of phloretin and naringenin. The promoter activity of acrR, encoding a regulatory protein involved in acrAB expression, was increased by naringenin. In planta, an induction of acrAB was proved by confocal laser scanning microscopy. Our results strongly suggest that the AcrAB transport system plays an important role as a protein complex required for virulence of E. amylovora in resistance toward apple phytoalexins and that it is required for successful colonization of a host plant.

  3. Transcriptomic responses of the olive fruit fly Bactrocera oleae and its symbiont Candidatus Erwinia dacicola to olive feeding

    Science.gov (United States)

    Pavlidi, Nena; Gioti, Anastasia; Wybouw, Nicky; Dermauw, Wannes; Ben-Yosef, Michael; Yuval, Boaz; Jurkevich, Edouard; Kampouraki, Anastasia; van Leeuwen, Thomas; Vontas, John

    2017-02-01

    The olive fruit fly, Bactrocera oleae, is the most destructive pest of olive orchards worldwide. The monophagous larva has the unique capability of feeding on olive mesocarp, coping with high levels of phenolic compounds and utilizing non-hydrolyzed proteins present, particularly in the unripe, green olives. On the molecular level, the interaction between B. oleae and olives has not been investigated as yet. Nevertheless, it has been associated with the gut obligate symbiotic bacterium Candidatus Erwinia dacicola. Here, we used a B.oleae microarray to analyze the gene expression of larvae during their development in artificial diet, unripe (green) and ripe (black) olives. The expression profiles of Ca. E. dacicola were analyzed in parallel, using the Illumina platform. Several genes were found overexpressed in the olive fly larvae when feeding in green olives. Among these, a number of genes encoding detoxification and digestive enzymes, indicating a potential association with the ability of B. oleae to cope with green olives. In addition, a number of biological processes seem to be activated in Ca. E. dacicola during the development of larvae in olives, with the most notable being the activation of amino-acid metabolism.

  4. Structural Characterization of Core Region in Erwinia amylovora Lipopolysaccharide.

    Science.gov (United States)

    Casillo, Angela; Ziaco, Marcello; Lindner, Buko; Merino, Susana; Mendoza-Barberá, Elena; Tomás, Juan M; Corsaro, Maria Michela

    2017-03-04

    Erwinia amylovora (E. amylovora) is the first bacterial plant pathogen described and demonstrated to cause fire blight, a devastating plant disease affecting a wide range of species including a wide variety of Rosaceae. In this study, we reported the lipopolysaccharide (LPS) core structure from E. amylovora strain CFBP1430, the first one for an E. amylovora highly pathogenic strain. The chemical characterization was performed on the mutants waaL (lacking only the O-antigen LPS with a complete LPS-core), wabH and wabG (outer-LPS core mutants). The LPSs were isolated from dry cells and analyzed by means of chemical and spectroscopic methods. In particular, they were subjected to a mild acid hydrolysis and/or a hydrazinolysis and investigated in detail by one and two dimensional Nuclear Magnetic Resonance (NMR) spectroscopy and ElectroSpray Ionization Fourier Transform-Ion Cyclotron Resonance (ESI FT-ICR) mass spectrometry.

  5. Structural Characterization of Core Region in Erwinia amylovora Lipopolysaccharide

    Directory of Open Access Journals (Sweden)

    Angela Casillo

    2017-03-01

    Full Text Available Erwinia amylovora (E. amylovora is the first bacterial plant pathogen described and demonstrated to cause fire blight, a devastating plant disease affecting a wide range of species including a wide variety of Rosaceae. In this study, we reported the lipopolysaccharide (LPS core structure from E. amylovora strain CFBP1430, the first one for an E. amylovora highly pathogenic strain. The chemical characterization was performed on the mutants waaL (lacking only the O-antigen LPS with a complete LPS-core, wabH and wabG (outer-LPS core mutants. The LPSs were isolated from dry cells and analyzed by means of chemical and spectroscopic methods. In particular, they were subjected to a mild acid hydrolysis and/or a hydrazinolysis and investigated in detail by one and two dimensional Nuclear Magnetic Resonance (NMR spectroscopy and ElectroSpray Ionization Fourier Transform-Ion Cyclotron Resonance (ESI FT-ICR mass spectrometry.

  6. Structural Characterization of Core Region in Erwinia amylovora Lipopolysaccharide

    Science.gov (United States)

    Casillo, Angela; Ziaco, Marcello; Lindner, Buko; Merino, Susana; Mendoza-Barberá, Elena; Tomás, Juan M.; Corsaro, Maria Michela

    2017-01-01

    Erwinia amylovora (E. amylovora) is the first bacterial plant pathogen described and demonstrated to cause fire blight, a devastating plant disease affecting a wide range of species including a wide variety of Rosaceae. In this study, we reported the lipopolysaccharide (LPS) core structure from E. amylovora strain CFBP1430, the first one for an E. amylovora highly pathogenic strain. The chemical characterization was performed on the mutants waaL (lacking only the O-antigen LPS with a complete LPS-core), wabH and wabG (outer-LPS core mutants). The LPSs were isolated from dry cells and analyzed by means of chemical and spectroscopic methods. In particular, they were subjected to a mild acid hydrolysis and/or a hydrazinolysis and investigated in detail by one and two dimensional Nuclear Magnetic Resonance (NMR) spectroscopy and ElectroSpray Ionization Fourier Transform-Ion Cyclotron Resonance (ESI FT-ICR) mass spectrometry. PMID:28273861

  7. Cloud Activation Characteristics of Airborne Erwinia carotovora Cells.

    Science.gov (United States)

    Franc, Gary D.; Demott, Paul J.

    1998-10-01

    Several strains of plant pathogenic bacteria, Erwinia carotovora carotovora and E. carotovora atroseptica, were observed to be active as cloud condensation nuclei (CCN). The CCN supersaturation spectra of bacterial aerosols were measured in the laboratory and compared to the activity of ammonium sulfate. Approximately 25%-30% of the aerosolized bacterial cells activated droplets at 1% water supersaturation compared to 80% activation of the ammonium sulfate aerosol. Physical and numerical simulations of cloud droplet activation and growth on bacteria were also performed. Both simulations predict that aerosolized bacteria will be incorporated into cloud droplets during cloud formation. Results strongly support the hypothesis that significant numbers of the tested bacterial strains are actively involved in atmospheric cloud formation and precipitation processes following natural aerosolization and vertical transport to cloud levels.

  8. Immobilization of glucosyltransferase from Erwinia sp. using two different techniques.

    Science.gov (United States)

    Contesini, Fabiano Jares; Ibarguren, Carolina; Grosso, Carlos Raimundo Ferreira; Carvalho, Patrícia de Oliveira; Sato, Hélia Harumi

    2012-04-15

    Two different techniques of glucosyltransferase immobilization were studied for the conversion of sucrose into isomaltulose. The optimum conditions for immobilization of Erwinia sp. glucosyltransferase onto Celite 545, determined using response surface methodology, was pH 4.0 and 170 U of glucosyltransferase/g of Celite 545. Using this conditions more than 60% conversion of sucrose into isomaltulose can be obtained. The immobilization of glucosyltransferase was also studied by its entrapment in microcapsules of low-methoxyl pectin and fat (butter and oleic acid). The non-lyophilized microcapsules of pectin, containing the enzyme and fat, showed higher glucosyltransferase activity, compared with lyophilized microcapsules containing enzyme plus fat, and also lyophilized microcapsules containing enzyme without fat addition. The non-lyophilized microcapsules of pectin containing the glucosyltransferase and fat, converted 30% of sucrose into isomaltulose in the first batch. However the conversion decreased to 5% at the 10th batch, indicating inactivation of the enzyme.

  9. Complete genome sequences of three Erwinia amylovora phages isolated in north america and a bacteriophage induced from an Erwinia tasmaniensis strain.

    Science.gov (United States)

    Müller, I; Kube, M; Reinhardt, R; Jelkmann, W; Geider, K

    2011-02-01

    Fire blight, a plant disease of economic importance caused by Erwinia amylovora, may be controlled by the application of bacteriophages. Here, we provide the complete genome sequences and the annotation of three E. amylovora-specific phages isolated in North America and genomic information about a bacteriophage induced by mitomycin C treatment of an Erwinia tasmaniensis strain that is antagonistic for E. amylovora. The American phages resemble two already-described viral genomes, whereas the E. tasmaniensis phage displays a singular genomic sequence in BLAST searches.

  10. Genomics of iron acquisition in the plant pathogen Erwinia amylovora: insights in the biosynthetic pathway of the siderophore desferrioxamine E.

    Science.gov (United States)

    Smits, Theo H M; Duffy, Brion

    2011-10-01

    Genomics has clarified the biosynthetic pathway for desferrioxamine E critical for iron acquisition in the enterobacterial fire blight pathogen Erwinia amylovora. Evidence for each of the individual steps and the role of desferrioxamine E biosynthesis in pathogen virulence and cell protection from host defenses is presented. Using comparative genomics, it can be concluded that desferrioxamine biosynthesis is ancestral within the genera Erwinia and Pantoea.

  11. Detection of soft rot Erwinia spp. on seed potatoes: conductimetry versus dilution plating, PCR and serological assays

    NARCIS (Netherlands)

    Fraaije, B.A.; Appels, M.; Boer, de S.H.; Vuurde, van J.W.L.; Bulk, van den R.W.

    1997-01-01

    Automated conductance measurements in polypectate medium were used for the detection of pathogenic soft rot Erwinia spp. in potato peel extracts. The detection threshold for Erwinia carotovora subsp. atroseptica (Eca) in inoculated peel extracts was ca. 104 colony forming units (cfu) ml-1 when sampl

  12. Identification of Erwinia species isolated from apples and pears by differential PCR.

    Science.gov (United States)

    Gehring, I; Geider, K

    2012-04-01

    Many pathogenic and epiphytic bacteria isolated from apples and pears belong to the genus Erwinia; these include the species E. amylovora, E. pyrifoliae, E. billingiae, E. persicina, E. rhapontici and E. tasmaniensis. Identification and classification of freshly isolated bacterial species often requires tedious taxonomic procedures. To facilitate routine identification of Erwinia species, we have developed a PCR method based on species-specific oligonucleotides (SSOs) from the sequences of the housekeeping genes recA and gpd. Using species-specific primers that we report here, differentiation was done with conventional PCR (cPCR) and quantitative PCR (qPCR) applying two consecutive primer annealing temperatures. The specificity of the primers depends on terminal Single Nucleotide Polymorphisms (SNPs) that are characteristic for the target species. These PCR assays enabled us to distinguish eight Erwinia species, as well as to identify new Erwinia isolates from plant surfaces. When performed with mixed bacterial cultures, they only detected a single target species. This method is a novel approach to classify strains within the genus Erwinia by PCR and it can be used to confirm other diagnostic data, especially when specific PCR detection methods are not already available. The method may be applied to classify species within other bacterial genera.

  13. Host exopolysaccharide quantity and composition impact Erwinia amylovora bacteriophage pathogenesis.

    Science.gov (United States)

    Roach, Dwayne R; Sjaarda, David R; Castle, Alan J; Svircev, Antonet M

    2013-05-01

    Erwinia amylovora bacteriophages (phages) belonging to the Myoviridae and Podoviridae families demonstrated a preference for either high-exopolysaccharide-producing (HEP) or low-exopolysaccharide-producing (LEP) bacterial hosts when grown on artificial medium without or with sugar supplementation. Myoviridae phages produced clear plaques on LEP hosts and turbid plaques on HEP hosts. The reverse preference was demonstrated by most Podoviridae phages, where clear plaques were seen on HEP hosts. Efficiency of plating (EOP) was determined by comparing phage growth on the original isolation host to the that on the LEP or HEP host. Nine of 10 Myoviridae phages showed highest EOPs on LEP hosts, and 8 of 11 Podoviridae phages had highest EOPs on HEP hosts. Increasing the production of EPS on sugar-supplemented medium or decreasing production by knocking out the synthesis of amylovoran or levan, the two EPSs produced by E. amylovora, indicated that these components play crucial roles in phage infection. Amylovoran was virtually essential for proliferation of most Podoviridae phages when phage population growth was compared to the wild type. Decreased levan production resulted in a significant reduction of progeny from phages in the Myoviridae family. Thus, Podoviridae phages are adapted to hosts that produce high levels of exopolysaccharides and are dependent on host-produced amylovoran for pathogenesis. Myoviridae phages are adapted to hosts that produce lower levels of exopolysaccharides and host-produced levan.

  14. RHEOLOGICAL BEHAVIOR OF ERWINIA GUM IN AQUEOUS SOLUTION

    Institute of Scientific and Technical Information of China (English)

    Li-na Zhang; Mei Zhang; Jing-hua Chen; Hideki Iijima; Hiromichi Tsuchiya

    1999-01-01

    Erwinia (E) gum, an extracellular polysaccharide, is composed of fucose, galatose, glucose and glucuronic acid. Its viscosity behavior was investigated by a low-shear-rate multiball viscometer and a rotational viscometer. Its weight-average molecular weight Mw and intrinsic viscosity [η] in 0.2 mol/L NaCl aqueous solution were measured by light scattering method at 35℃ and viscometry at 25℃ and found to be 1.06 × 106 g/mol and 1050 mL/g, respectively, and its aggregates in aqueous solution were proved by gel permeation chromatography (GPC). These results indicated that E gum in water has exceedingly high viscosity and exhibits Binham fluid behavior, owing to its aggregation. The viscosity of E gum decreased with increasing temperature, and the turning point appeared at 38℃ for dilute solution and 80℃ for concentrated solution suggesting that the aggregates of E gum in water started to disaggregate under these temperatures. In addition, the aggregates can be disrupted by adding either acid or base. The experimental results indicated that the E gum is a good thickening agent, and its fluid behavior is similar to xanthan.

  15. Differential Colonization Dynamics of Cucurbit Hosts by Erwinia tracheiphila.

    Science.gov (United States)

    Vrisman, Cláudio M; Deblais, Loïc; Rajashekara, Gireesh; Miller, Sally A

    2016-07-01

    Bacterial wilt is one of the most destructive diseases of cucurbits in the Midwestern and Northeastern United States. Although the disease has been studied since 1900, host colonization dynamics remain unclear. Cucumis- and Cucurbita-derived strains exhibit host preference for the cucurbit genus from which they were isolated. We constructed a bioluminescent strain of Erwinia tracheiphila (TedCu10-BL#9) and colonization of different cucurbit hosts was monitored. At the second-true-leaf stage, Cucumis melo plants were inoculated with TedCu10-BL#9 via wounded leaves, stems, and roots. Daily monitoring of colonization showed bioluminescent bacteria in the inoculated leaf and petiole beginning 1 day postinoculation (DPI). The bacteria spread to roots via the stem by 2 DPI, reached the plant extremities 4 DPI, and the plant wilted 6 DPI. However, Cucurbita plants inoculated with TedCu10-BL#9 did not wilt, even at 35 DPI. Bioluminescent bacteria were detected 6 DPI in the main stem of squash and pumpkin plants, which harbored approximately 10(4) and 10(1) CFU/g, respectively, of TedCu10-BL#9 without symptoms. Although significantly less systemic plant colonization was observed in nonpreferred host Cucurbita plants compared with preferred hosts, the mechanism of tolerance of Cucurbita plants to E. tracheiphila strains from Cucumis remains unknown.

  16. Tasmancin and lysogenic bacteriophages induced from Erwinia tasmaniensis strains.

    Science.gov (United States)

    Müller, Ina; Lurz, Rudi; Geider, Klaus

    2012-07-25

    Mitomycin C treatment of Erwinia tasmaniensis strains from Australia induced prophages and the expression of bacteriocins. The bacteriocin named tasmancin inhibited E. tasmaniensis strains from South Africa and Germany. A gene cluster with a klebicin-related operon and an immunity protein was detected on plasmid pET46 from E. tasmaniensis strain Et1/99. PCR reactions using primers directed to this region produced signals for several strains originating from Australia, but not for strains isolated in South Africa and Germany. The latter isolates lacked plasmid pET46. Bacteriophages were induced from E. tasmaniensis strains Et88 and Et14/99, both isolates from South-Eastern Australia. These phages formed plaques on several other strains from this region, as well as on E. tasmaniensis strains from South Africa and Germany. Sequencing revealed similarity of phages ϕEt88 and ϕEt14, which shared the host range on E. tasmaniensis strains. Bacteriophages and tasmancin may interfere with the viability of several related E. tasmaniensis strains in the environment of carrier strains.

  17. Erwinia piriflorinigrans sp. nov., a novel pathogen that causes necrosis of pear blossoms.

    Science.gov (United States)

    López, María M; Roselló, Montserrat; Llop, Pablo; Ferrer, Sergi; Christen, Richard; Gardan, Louis

    2011-03-01

    Eight Erwinia strains, isolated from necrotic pear blossoms in València, Spain, were compared with reference strains of Erwinia amylovora and Erwinia pyrifoliae, both of which are pathogenic to species of pear tree, and to other species of the family Enterobacteriaceae using a polyphasic approach. Phenotypic analyses clustered the novel isolates into one phenon, distinct from other species of the genus Erwinia, showing that the novel isolates constituted a homogeneous phenotypic group. Rep-PCR profiles, PCR products obtained with different pairs of primers and plasmid contents determined by restriction analysis showed differences between the novel strains and reference strains of E. amylovora and E. pyrifoliae. Phylogenetic analysis of 16S rRNA, gpd and recA gene sequences showed that the eight novel strains could not be assigned to any recognized species. On the basis of DNA-DNA hybridization studies, the novel isolates constituted a single group with relatedness values of 87-100  % to the designated type strain of the group, CFBP 5888(T). Depending on the method used, strain CFBP 5888(T) showed DNA-DNA relatedness values of between 22.7 and 50  % to strains of the closely related species E. amylovora and E. tasmaniensis. The DNA G+C contents of two of the novel strains, CFBP 5888(T) and CFBP 5883, were 51.1 and 50.5 mol%, respectively. On the basis of these and previous results, the novel isolates represent a novel species of the genus Erwinia, for which the name Erwinia piriflorinigrans sp. nov. is proposed. The type strain is CFBP 5888(T) (=CECT 7348(T)).

  18. Functional assembly of the foreign carotenoid lycopene into the photosynthetic apparatus of Rhodobacter sphaeroides, achieved by replacement of the native 3-step phytoene desaturase with its 4-step counterpart from Erwinia herbicola.

    Science.gov (United States)

    Garcia-Asua, Guillermo; Cogdell, Richard J; Hunter, C Neil

    2002-04-01

    Photosynthetic organisms synthesize a diverse range of carotenoids. These pigments are important for the assembly, function and stability of photosynthetic pigment-protein complexes, and they are used to quench harmful radicals. The photosynthetic bacterium Rhodobacter sphaeroides was used as a model system to explore the origin of carotenoid diversity. Replacing the native 3-step phytoene desaturase (CrtI) with the 4-step enzyme from Erwinia herbicola results in significant flux down the spirilloxanthin pathway for the first time in Rb. sphaeroides. In Rb. sphaeroides, the completion of four desaturations to lycopene by the Erwinia CrtI appears to require the absence of CrtC and, in a crtC background, even the native 3-step enzyme can synthesize a significant amount (13%) of lycopene, in addition to the expected neurosporene. We suggest that the CrtC hydroxylase can intervene in the sequence of reactions catalyzed by phytoene desaturase. We investigated the properties of the lycopene-synthesizing strain of Rb. sphaeroides. In the LH2 light-harvesting complex, lycopene transfers absorbed light energy to the bacteriochlorophylls with an efficiency of 54%, which compares favourably with other LH2 complexes that contain carotenoids with 11 conjugated double bonds. Thus, lycopene can join the assembly pathway for photosynthetic complexes in Rb. sphaeroides, and can perform its role as an energy donor to bacteriochlorophylls.

  19. Erwinia amylovora affects the phenylpropanoid-flavonoid pathway in mature leaves of Pyrus communis cv. Conférence.

    Science.gov (United States)

    Vrancken, K; Holtappels, M; Schoofs, H; Deckers, T; Treutter, D; Valcke, R

    2013-11-01

    Flavonoids, which are synthesized by the phenylpropanoid-flavonoid pathway, not only contribute to fruit colour and photoprotection, they also may provide antimicrobial and structural components during interaction with micro-organisms. A possible response of this pathway was assessed in both mature and immature leaves of shoots of 2-year-old pear trees cv. Conférence, which were inoculated with the gram-negative bacterium Erwinia amylovora strain SGB 225/12, were mock-inoculated or were left untreated. The phenylpropanoid-flavonoid pathway was analysed by histological studies, by gene expression using RT-qPCR and by HPLC analyses of the metabolites at different time intervals after infection. Transcription patterns of two key genes anthocyanidin reductase (ANR) and chalcone synthase (CHS) related to the phenylpropanoid-flavonoid pathway showed differences between control, mock-inoculated and E. amylovora-inoculated mature leaves, with the strongest reaction 48 h after inoculation. The impact of E. amylovora was also visualised in histological sections, and confirmed by HPLC, as epicatechin -which is produced via ANR- augmented 72 h after inoculation in infected leaf tissue. Besides the effect of treatments, ontogenesis-related differences were found as well. The increase of certain key genes, the rise in epicatechin and the visualisation in several histological sections in this study suggest a non-negligible impact on the phenylpropanoid-flavonoid pathway in Pyrus communis due to inoculation with E. amylovora. In this study, we propose a potential role of this pathway in defence mechanisms, providing a detailed analysis of the response of this system attributable to inoculation with E. amylovora.

  20. Conventional and real-time PCRs for detection of Erwinia piriflorinigrans allow its distinction from the fire blight pathogen, Erwinia amylovora.

    Science.gov (United States)

    Barbé, Silvia; Bertolini, Edson; Roselló, Montserrat; Llop, Pablo; López, María M

    2014-04-01

    Erwinia piriflorinigrans is a new pathogenic species of the bacterial genus Erwinia that has been described recently in Spain. Accurate detection and identification of E. piriflorinigrans are challenging because its symptoms on pear blossoms are similar to those caused by Erwinia amylovora, the causal agent of fire blight. Moreover, these two species share phenotypic and molecular characteristics. Two specific and sensitive conventional and real-time PCR protocols were developed to identify and detect E. piriflorinigrans and to differentiate it from E. amylovora and other species of this genus. These protocols were based on sequences from plasmid pEPIR37, which is present in all strains of E. piriflorinigrans analyzed. After the stability of the plasmid was demonstrated, the specificities of the protocols were confirmed by the amplification of all E. piriflorinigrans strains tested, whereas 304 closely related pathogenic and nonpathogenic Erwinia strains and microbiota from pear trees were not amplified. In sensitivity assays, 10(3) cells/ml extract were detected in spiked plant material by conventional or real-time PCR, and 10(2) cells/ml were detected in DNA extracted from spiked plant material by real-time PCR. The protocols developed here succeeded in detecting E. piriflorinigrans in 102 out of 564 symptomatic and asymptomatic naturally infected pear samples (flowers, cortex stem tissue, leaves, shoots, and fruitlets), in necrotic Pyracantha sp. blossoms, and in necrotic pear and apple tissues infected with both E. amylovora and E. piriflorinigrans. Therefore, these new tools can be used in epidemiological studies that will enhance our understanding of the life cycle of E. piriflorinigrans in different hosts and plant tissues and its interaction with E. amylovora.

  1. Development of a Duplex PCR for Identification of Erwinia amylovora%步双重PCR法检测梨火疫病原细菌(Erwinia amylovora)

    Institute of Scientific and Technical Information of China (English)

    许景升; 徐进; 冯洁

    2008-01-01

    @@ 由梨火疫病原细菌(Erwinia amylovom)导致的火疫病(fire blight)是梨、苹果及其它蔷薇科植物上的毁灭性病害,我国将其定为对外一类检疫性有害生物.该病随种苗、果实及包装材料传播.

  2. Complete genome sequence of the plant pathogen Erwinia amylovora strain ATCC 49946

    Science.gov (United States)

    Erwinia amylovora causes the economically important disease fire blight that affects rosaceous plants, especially pear and apple. Here we report the complete genome sequence and annotation of strain ATCC 49946. The analysis of the sequence and its comparison with sequenced genomes of closely related...

  3. Morphological and biochemical characterization of Erwinia amylovora-induced hypersensitive cell death in apple leaves

    NARCIS (Netherlands)

    Iakimova, E.T.; Sobiczewski, P.; Michalczuk, L.; Wegrzynowicz-Lesiak, E.; Mikicinski, A.; Woltering, E.J.

    2013-01-01

    In attached apple leaves, spot-inoculated with Erwinia amylovora, the phenotypic appearance of the hypersensitive response (HR) and the participation of ethylene, reactive oxygen species (ROS) and of vacuolar processing enzyme (VPE) (a plant caspase-1-like protease) were analysed. The HR in both the

  4. Plasmid content of Erwinia amylovora in orchards in Washington and Oregon

    Science.gov (United States)

    We examined the plasmid content of a collection of 305 isolates of Erwinia amylovora from Washington and Oregon in the Pacific Northwest of the United States with PCR assays and RFLP. Nearly all isolates of E. amylovora carried plasmid pEA29, which is not found in other species of bacteria, but 4% ...

  5. Plasmid Content of Isolates of Erwinia amylovora from Orchards in Washington and Oregon in the USA

    Science.gov (United States)

    Washington (WA) and Oregon (OR) represent a major pome fruit production region of the United States, and streptomycin-resistant isolates of the fire blight pathogen Erwinia amylovora are common in orchards in this region. We examined the plasmid content of a collection of more than 200 isolates of ...

  6. Antibiosis by Pantoea agglomerans biocontrol strain E325 against Erwinia amylovora on apple blossom stigmas

    Science.gov (United States)

    Pantoea agglomerans E325, the active ingredient in a commercial product for fire blight control, was previously shown in vitro to produce a unique alkaline- and phosphate-sensitive antibiotic specific to Erwinia amylovora. Antibiosis was evaluated as a mode of antagonism on blossom stigmas using two...

  7. Erwinia amylovora effector protein Eop1 suppresses PAMP-triggered immunity in Malus

    Science.gov (United States)

    Erwinia amylovora (Ea) utilizes a type three secretion system (T3SS) to deliver effector proteins into plant host cells. Several Ea effectors have been identified based on their sequence similarity to plant and animal bacterial pathogen effectors; however, the function of the majority of Ea effecto...

  8. Using functional genomics to identify molecular markers for fire blight resistance (Erwinia amylovora) in apple (Malus)

    Science.gov (United States)

    Fire blight, caused by Erwinia amylovora (Ea), is a destructive disease of apple (Malus), pear (Pyrus) and some woody ornamentals in the rose family (Rosaceae). The goal of this project is to use a functional genomics approach to develop tools to breed fire blight resistant apples. Six hundred fifty...

  9. Eerste jaar Erwinia-project legt topje van de ijsberg bloot

    NARCIS (Netherlands)

    Velvis, H.; Wolf, van der J.M.

    2006-01-01

    Over de herkomst van de Erwinia bacterie heerst nog altijd veel onduidelijkheid. Daarom is een onderzoek naar deze bacterie gestart. In vier jaar tijd worden diverse aardappelrassen onderzocht in de verschillende stadia van teelt en opslag. Op deze manier hoopt men te achterhalen waar de herkomst va

  10. The presence and survival of soft rot (Erwinia) in flower bulb production systems

    NARCIS (Netherlands)

    Doorn, van J.; Vreeburg, P.J.M.; Leeuwen, van P.J.; Dees, R.H.L.

    2011-01-01

    Soft rot is causing increasing damage in the flower bulb industry. Bulbous ornamentals such as Hyacinthus, Dahlia, Iris, Muscari, Freesia and Zantedeschia can be infected. Soft rot in flower bulbs is mainly caused by Dickeya spp. (Dickeya spp.) and Erwinia carotovora subsp. carotovora (Pectobacteriu

  11. Evaluation of a PCR kit for the detection of Erwinia carotovora subsp. atroseptica on potato tubers

    NARCIS (Netherlands)

    Frechon, D.; Exbrayat, P.; Helias, V.; Hyman, L.J.; Jouan, B.; Llop, P.; Lopez, M.M.; Payet, N.; Perombelon, M.C.M.; Toth, I.K.; Beckhoven, van J.R.C.M.; Wolf, van der J.M.; Bertheau, Y.

    1998-01-01

    A PCR-based kit, Probelia(TM), for the detection of Erwinia carotovora subsp. atroseptica (Eca) on potatoes was evaluated at five laboratories in four countries. The kit is based on DNA-specific PCR amplification followed by detection of amplicons by hybridization to a peroxidase-labelled DNA probe

  12. Differential lysine acetylation profiles of Erwinia amylovora strains revealed by proteomics

    Science.gov (United States)

    Protein lysine acetylation (LysAc) in bacteria has recently been demonstrated to be widespread in E. coli and Salmonella and to broadly regulate bacterial physiology and metabolism. However, LysAc in plant pathogenic bacteria is largely unknown. Here we report the lysine acetylome of Erwinia amylovo...

  13. AmyR is a novel negative regulator of amylovoran production in Erwinia amylovora

    Science.gov (United States)

    We have previously reported the characterization of an orphan gene ybjN from Escherichia coli. In this study, we attempted to understand the role of amyR in Erwinia amylovora, a functionally conserved homolog of E. coli ybjN. As reported earlier, amylovoran production in the amyR knockout mutant is ...

  14. Genome-wide identification of genes regulated by the Rcs Phosphorelay system in Erwinia amylovora

    Science.gov (United States)

    The exopolysaccharide amylovoran is one of the major pathgenicity factors in Erwinia amylovora, the causal agent of fire blight of apples and pears. We have previously demonstrated that the RcsBCD phosphorelay system is essential for virulence by controlling amylovoran biosynthesis. We have also fou...

  15. Draft genome sequence of Erwinia tracheiphila, an economically important bacterial pathogen of cucurbits

    Science.gov (United States)

    Erwinia tracheiphila is one of the most economically important pathogen of cucumbers, melons, squashes, pumpkins, and gourds, in the Northeastern and Midwestern United States, yet the molecular pathology remains uninvestigated. Here we report the first draft genome sequence of an E. tracheiphila str...

  16. [Erwinia amylovora--the fire blight pathogen of trees in Ukraine].

    Science.gov (United States)

    Iakovleva, L M; Moroz, S N; Shcherbina, T N; Ogorodnik, L E; Gvozdiak, R I; Patyka, V F

    2014-01-01

    Niduses of fire blight of fruit and ornamental trees have been found in the Kyiv and Vinnitsa regions of Ukraine. Pathogen Erwinia amylovora was isolated between April and October. The pathogen was often accompanied by bacteria Pseudomonas syringae pv. syringae. Artificial infection with a mixture of bacteria E. amylovora and P. syringae pv. syringae accelerates and enhances the disease process in the laboratory.

  17. Genetic islands in pome fruit pathogenic and nonpathogenic Erwinia species and related plasmids

    Directory of Open Access Journals (Sweden)

    Pablo eLlop

    2015-08-01

    Full Text Available New pathogenic bacteria species belonging to the genus Erwinia associated with pome fruit trees (Erwinia pyrifoliae, E. piriflorinigrans, E. uzenensis have been increasingly described in the last years, and comparative analyses have found that all these species share several genetic characteristics. Studies at different level (whole genome comparison, virulence genes, plasmid content, etc. show a high intraspecies homogeneity (i.e. among E. amylovora strains and also abundant similarities appear between the different Erwinia species: presence of plasmids of similar size in the pathogenic species; high similarity in several genes associated with exopolysaccharide production and hence, with virulence, as well as in some other genes, in the chromosomes. Many genetic similarities have been observed also among some of the plasmids (and genomes from the pathogenic species and E. tasmaniensis or E. billingiae, two epiphytic species on the same hosts. The amount of genetic material shared in this genus varies from individual genes to clusters, genomic islands and genetic material that even may constitute a whole plasmid. Recent research on evolution of erwinias point out the horizontal transfer acquisition of some genomic islands that were subsequently lost in some species and several pathogenic traits that are still present. How this common material has been obtained and is efficiently maintained in different species belonging to the same genus sharing a common ecological niche provides an idea of the origin and evolution of the pathogenic Erwinia and the interaction with nonpathogenic species present in the same niche, and the role of the genes that are conserved in all of them.

  18. Genetic islands in pome fruit pathogenic and non-pathogenic Erwinia species and related plasmids.

    Science.gov (United States)

    Llop, Pablo

    2015-01-01

    New pathogenic bacteria belonging to the genus Erwinia associated with pome fruit trees (Erwinia, E. piriflorinigrans, E. uzenensis) have been increasingly described in the last years, and comparative analyses have found that all these species share several genetic characteristics. Studies at different level (whole genome comparison, virulence genes, plasmid content, etc.) show a high intraspecies homogeneity (i.e., among E. amylovora strains) and also abundant similarities appear between the different Erwinia species: presence of plasmids of similar size in the pathogenic species; high similarity in several genes associated with exopolysaccharide production and hence, with virulence, as well as in some other genes, in the chromosomes. Many genetic similarities have been observed also among some of the plasmids (and genomes) from the pathogenic species and E. tasmaniensis or E. billingiae, two epiphytic species on the same hosts. The amount of genetic material shared in this genus varies from individual genes to clusters, genomic islands and genetic material that even may constitute a whole plasmid. Recent research on evolution of erwinias point out the horizontal transfer acquisition of some genomic islands that were subsequently lost in some species and several pathogenic traits that are still present. How this common material has been obtained and is efficiently maintained in different species belonging to the same genus sharing a common ecological niche provides an idea of the origin and evolution of the pathogenic Erwinia and the interaction with non-pathogenic species present in the same niche, and the role of the genes that are conserved in all of them.

  19. Partial resistance to Erwinia carotovora SUBSP. carotovora and plant vigour among F1 hybrids of Zantedeschia cultivars

    NARCIS (Netherlands)

    Snijder, R.C.; Tuyl, van J.M.

    2004-01-01

    The potential of breeding Zantedeschia cultivars for resistance to soft rot caused by Erwinia carotovora subsp. carotovora (syn. Pectobacterium carotovorum subsp. carotovorum) was determined. Hybrids of six cultivars (`Back Magic`, Galaxy`, Pink Persuasion`, Sensation`, `Treasure` and `Florex Gold`)

  20. 菊花破壁饮片的HPLC指纹图谱研究%Study on HPLC Fingerprint of Cell Wall-Broken Decoction Pieces of Chrysanthemi Flos

    Institute of Scientific and Technical Information of China (English)

    刘星云; 王慧玲; 彭丽华; 成金乐

    2016-01-01

    目的:建立菊花破壁饮片HPLC指纹图谱,并分析破壁饮片成品与其中间品、原料的化学成分相关性,为菊花破壁饮片整体质量评价提供依据.方法:采用高效液相色谱法,十八烷基硅烷键合硅胶为填充剂的色谱柱(250 mm×4.6 mm,5μm),以乙腈-0.5%磷酸溶液为流动相进行梯度洗脱,检测波长:348 nm,柱温:35℃.结果:建立了菊花破壁饮片的HPLC指纹图谱,得到了15个共有特征峰,11批样品的相似度达0.98以上,方法学考察结果符合指纹图谱技术要求.结论:所建立的方法稳定、可靠、重复性好,可用于菊花破壁饮片质量控制和综合评价.%Objective:To establish the HPLC fingerprint of Cell Wall-Broken Decoction Pieces of Chrysanthemi Flos, and analyze the relativity between its decoction pieces,broken powder and broken particle,helping to evaluate its quality comprehensively.Methods:RP-HPLC method was performed on an Agilent C18 (250 mm ×4.6 mm,5 μm)column with a gradient elution composed of acetonitrile-aqueous solution containing 0.5% phosphoric acid.The column temperature was set at 35 ℃,while the detective wavelength was set at 348 nm.Results:The chromatographic fingerprint common pattern was established.Fifteen mutual peaks were obtained from the chromatograms of eleven batches of samples.Conclusion:The method with good reproducibility is reliable and stable,which is feasible for quality control of cell wall-broken decoction pieces of Chrysanthemi Flos.

  1. The crystal structure of Erwinia amylovora levansucrase provides a snapshot of the products of sucrose hydrolysis trapped into the active site.

    Science.gov (United States)

    Wuerges, Jochen; Caputi, Lorenzo; Cianci, Michele; Boivin, Stephane; Meijers, Rob; Benini, Stefano

    2015-09-01

    Levansucrases are members of the glycoside hydrolase family and catalyse both the hydrolysis of the substrate sucrose and the transfer of fructosyl units to acceptor molecules. In the presence of sufficient sucrose, this may either lead to the production of fructooligosaccharides or fructose polymers. Aim of this study is to rationalise the differences in the polymerisation properties of bacterial levansucrases and in particular to identify structural features that determine different product spectrum in the levansucrase of the Gram-negative bacterium Erwinia amylovora (Ea Lsc, EC 2.4.1.10) as compared to Gram-positive bacteria such as Bacillus subtilis levansucrase. Ea is an enterobacterial pathogen responsible for the Fire Blight disease in rosaceous plants (e.g., apple and pear) with considerable interest for the agricultural industry. The crystal structure of Ea Lsc was solved at 2.77 Å resolution and compared to those of other fructosyltransferases from Gram-positive and Gram-negative bacteria. We propose the structural features, determining the different reaction products, to reside in just a few loops at the rim of the active site funnel. Moreover we propose that loop 8 may have a role in product length determination in Gluconacetobacter diazotrophicus LsdA and Microbacterium saccharophilum FFase. The Ea Lsc structure shows for the first time the products of sucrose hydrolysis still bound in the active site.

  2. HopX1 in Erwinia amylovora functions as an avirulence protein in apple and is regulated by HrpL.

    Science.gov (United States)

    Bocsanczy, A M; Schneider, D J; DeClerck, G A; Cartinhour, S; Beer, S V

    2012-02-01

    Fire blight is a devastating disease of rosaceous plants caused by the Gram-negative bacterium Erwinia amylovora. This pathogen delivers virulence proteins into host cells utilizing the type III secretion system (T3SS). Expression of the T3SS and of translocated and secreted substrates is activated by the alternative sigma factor HrpL, which recognizes hrp box promoters upstream of regulated genes. A collection of hidden Markov model (HMM) profiles was used to identify putative hrp boxes in the genome sequence of Ea273, a highly virulent strain of E. amylovora. Among potential virulence factors preceded by putative hrp boxes, two genes previously known as Eop3 and Eop2 were characterized. The presence of functionally active hrp boxes upstream of these two genes was confirmed by β-glucuronidase (GUS) assays. Deletion mutants of the latter candidate genes, renamed hopX1(Ea) and hopAK1(Ea), respectively, did not differ in virulence from the wild-type strain when assayed in pear fruit and apple shoots. The hopX1(Ea) deletion mutant of Ea273, complemented with a plasmid overexpressing hopX1(E)(a), suppressed the development of the hypersensitivity response (HR) when inoculated into Nicotiana benthamiana; however, it contributed to HR in Nicotiana tabacum and significantly reduced the progress of disease in apple shoots, suggesting that HopX1(Ea) may act as an avirulence protein in apple shoots.

  3. Carotenoid biosynthesis in bacteria: In vitro studies of a crt/bch transcription factor from Rhodobacter capsulatus and carotenoid enzymes from Erwinia herbicola

    Energy Technology Data Exchange (ETDEWEB)

    O`Brien, David Allen [Univ. of California, Berkeley, CA (United States)

    1992-11-01

    A putative transcription factor in Rhodobactor capsulatus which binds upstream of the crt and bch pigment biosynthesis operons and appears to play a role in the adaptation of the organism from the aerobic to the anaerobic-photosynthetic growth mode was characterized. Chapter 2 describes the identification of this factor through an in vitro mobility shift assay, as well as the determination of its binding properties and sequence specificity. Chapter 3 focuses on the isolation of this factor. Biochemistry of later carotenoid biosynthesis enzymes derived from the non-photosynthetic bacterium, Erwinia herbicola. Chapter 4 describes the separate overexpression and in vitro analysis of two enzymes involved in the main sequence of the carotenoid biosynthesis pathway, lycopene cyclase and 5-carotene hydroxylase. Chapter 5 examines the overexpression and enzymology of functionally active zeaxanthin glucosyltransferase, an enzyme which carries out a more unusual transformation, converting a carotenoid into its more hydrophilic mono- and diglucoside derivatives. In addition, amino acid homology with other glucosyltransferases suggests a putative binding site for the UDP-activated glucose substrate.

  4. Single Bacterium Detection Using Sers

    Science.gov (United States)

    Gonchukov, S. A.; Baikova, T. V.; Alushin, M. V.; Svistunova, T. S.; Minaeva, S. A.; Ionin, A. A.; Kudryashov, S. I.; Saraeva, I. N.; Zayarny, D. A.

    2016-02-01

    This work is devoted to the study of a single Staphylococcus aureus bacterium detection using surface-enhanced Raman spectroscopy (SERS) and resonant Raman spectroscopy (RS). It was shown that SERS allows increasing sensitivity of predominantly low frequency lines connected with the vibrations of Amide, Proteins and DNA. At the same time the lines of carotenoids inherent to this kind of bacterium are well-detected due to the resonance Raman scattering mechanism. The reproducibility and stability of Raman spectra strongly depend on the characteristics of nanostructured substrate, and molecular structure and size of the tested biological object.

  5. Regulatory network controlling extracellular proteins in Erwinia carotovora subsp. carotovora: FlhDC, the master regulator of flagellar genes, activates rsmB regulatory RNA production by affecting gacA and hexA (lrhA) expression.

    Science.gov (United States)

    Cui, Yaya; Chatterjee, Asita; Yang, Hailian; Chatterjee, Arun K

    2008-07-01

    Erwinia carotovora subsp. carotovora produces an array of extracellular proteins (i.e., exoproteins), including plant cell wall-degrading enzymes and Harpin, an effector responsible for eliciting hypersensitive reaction. Exoprotein genes are coregulated by the quorum-sensing signal, N-acyl homoserine lactone, plant signals, an assortment of transcriptional factors/regulators (GacS/A, ExpR1, ExpR2, KdgR, RpoS, HexA, and RsmC) and posttranscriptional regulators (RsmA, rsmB RNA). rsmB RNA production is positively regulated by GacS/A, a two-component system, and negatively regulated by HexA (PecT in Erwinia chrysanthemi; LrhA [LysR homolog A] in Escherichia coli) and RsmC, a putative transcriptional adaptor. While free RsmA, an RNA-binding protein, promotes decay of mRNAs of exoprotein genes, binding of RsmA with rsmB RNA neutralizes the RsmA effect. In the course of studies of GacA regulation, we discovered that a locus bearing strong homology to the flhDC operon of E. coli also controls extracellular enzyme production. A transposon insertion FlhDC(-) mutant produces very low levels of pectate lyase, polygalacturonase, cellulase, protease, and E. carotovora subsp. carotovora Harpin (Harpin(Ecc)) and is severely attenuated in its plant virulence. The production of these exoproteins is restored in the mutant carrying an FlhDC(+) plasmid. Sequence analysis and transcript assays disclosed that the flhD operon of E. carotovora subsp. carotovora, like those of other enterobacteria, consists of flhD and flhC. Complementation analysis revealed that the regulatory effect requires functions of both flhD and flhC products. The data presented here show that FlhDC positively regulates gacA, rsmC, and fliA and negatively regulates hexA (lrhA). Evidence shows that FlhDC controls extracellular protein production through cumulative effects on hexA and gacA. Reduced levels of GacA and elevated levels of HexA in the FlhDC(-) mutant are responsible for the inhibition of rsmB RNA

  6. Reaction of arracacha genotypes to the root soft rot caused by Pectobacterium chrysanthemi Reação de genótipos de mandioquinha-salsa à podridão-mole das raízes causada por Pectobacterium chrysanthemi

    Directory of Open Access Journals (Sweden)

    Gilmar Paulo Henz

    2005-01-01

    Full Text Available The purpose of this paper was to screen thirty-two arracacha genotypes for their reaction to root soft rot. Twenty roots of each genotype were inoculated with two Pectobacterium chrysanthemi isolates in a randomized experiment (10 roots/isolate. After inoculation, roots were individually wrapped with PVC film and kept at 26ºC in closed plastic bags. Soft rot lesions were recorded after 36 hours and genotypes were grouped in four classes of susceptibility by cluster analysis: 10 were less susceptible, 16 intermediate, 3 susceptible and 3 very susceptible. All the tested arracacha genotypes showed only variation in the degree of susceptibility.O objetivo deste trabalho foi avaliar a reação de 32 genótipos de mandioquinha-salsa à podridão-mole das raízes. Vinte raízes de cada genótipo foram inoculadas com dois isolados de Pectobacterium chrysanthemi em um experimento casualizado (10 raízes/isolado. Após a inoculação, as raízes foram embaladas com filmes de PVC e mantidas a 26ºC em sacos de plástico. As lesões de podridão-mole foram avaliadas após 36 horas e os genótipos agrupados em quatro classes de suscetibilidade por análise de agrupamento: 10 foram menos suscetíveis, 16 intermediários, 3 suscetíveis e 3 muito suscetíveis. Todos os genótipos avaliados demonstraram apenas variação no grau de suscetibilidade.

  7. Erwinia mallotivora sp., a new pathogen of papaya (Carica papaya) in Peninsular Malaysia.

    Science.gov (United States)

    Amin, Noriha Mat; Bunawan, Hamidun; Redzuan, Rohaiza Ahmad; Jaganath, Indu Bala S

    2010-12-24

    Erwinia mallotivora was isolated from papaya infected with dieback disease showing the typical symptoms of greasy, water-soaked lesions and spots on leaves. Phylogenetic analysis of 16S rRNA gene sequences showed that the strain belonged to the genus Erwinia and was united in a monophyletic group with E. mallotivora DSM 4565 (AJ233414). Earlier studies had indicated that the causal agent for this disease was E. papayae. However, our current studies, through Koch's postulate, have confirmed that papaya dieback disease is caused by E. mallotivora. To our knowledge, this is the first new discovery of E. mallotivora as a causal agent of papaya dieback disease in Peninsular Malaysia. Previous reports have suggested that E. mallotivora causes leaf spot in Mallotus japonicus. However, this research confirms it also to be pathogenic to Carica papaya.

  8. Erwinia mallotivora sp., a New Pathogen of Papaya (Carica papaya in Peninsular Malaysia

    Directory of Open Access Journals (Sweden)

    Noriha Mat Amin

    2010-12-01

    Full Text Available Erwinia mallotivora was isolated from papaya infected with dieback disease showing the typical symptoms of greasy, water-soaked lesions and spots on leaves. Phylogenetic analysis of 16S rRNA gene sequences showed that the strain belonged to the genus Erwinia and was united in a monophyletic group with E. mallotivora DSM 4565 (AJ233414. Earlier studies had indicated that the causal agent for this disease was E. papayae. However, our current studies, through Koch’s postulate, have confirmed that papaya dieback disease is caused by E. mallotivora. To our knowledge, this is the first new discovery of E. mallotivora as a causal agent of papaya dieback disease in Peninsular Malaysia. Previous reports have suggested that E. mallotivora causes leaf spot in Mallotus japonicus. However, this research confirms it also to be pathogenic to Carica papaya.

  9. Erwinia endophytica sp. nov., isolated from potato (Solanum tuberosum L.) stems.

    Science.gov (United States)

    Ramírez-Bahena, Martha Helena; Salazar, Sergio; Cuesta, Maria José; Tejedor, Carmen; Igual, Jose Mariano; Fernández-Pascual, Mercedes; Peix, Alvaro

    2015-12-04

    We analysed through a polyphasic taxonomy approach two bacterial strains coded BSTT30T and BSTT40, isolated in the course of a study of endophytic bacteria occurring in stems and roots of potato growing in a soil from Salamanca, Spain. The 16S rRNA gene sequence was identical in both strains and had 98.4% identity with respect to the closest relatives Erwinia tasmaniensis Et1/99T and E. rhapontici ATCC29283T and the following closest related species with 98.2 % similarity were E. billingiae E63T and E. toletana A37T, for which the strains were classified within genus Erwinia. The analysis of housekeeping genes gpd, gyrB and rpoD confirmed their phylogenetic affiliation and showed identities lower than 90% in all cases with respect to the mentioned closest relatives. The respiratory quinone is Q8. The major fatty acids are C16:0, C16:1 7c/ 16:1 6c in summed feature 3 and C18:1 7c/ 18:2 6,9c in summed feature 8. Oxidase negative and catalase positive. Glucose is fermented without gas production. Arginine dihydrolase, urease and indole production are negative. It can grow at 35ºC and at pH 10. The DNA G+C content was 50.1 mol %. DNA-DNA hybridization results showed values lower than 29% relatedness with respect to the type strains of the four closest related species. Therefore, the combined results of genotypic, phenotypic and chemotaxonomic data support the classification of strains BSTT30T and BSTT40 into a novel species of Erwinia, for which the name Erwinia endophytica sp. nov. is proposed. The type strain is BSTT30T (=LMG 28457T, CECT 8692T).

  10. Estimation of resistance pear cultivars to Erwinia amylovora using artificial immature pear fruits method

    OpenAIRE

    Gavrilović, Veljko; Stanisavljević, Rade; Stošić, Stefan; Stevanović, Miloš; Aleksić, Goran; Stajić, Milica; Dolovac, Nenad

    2014-01-01

    Susceptibility of different pear cultivars to Erwinia amaylovora by artificial inoculated immature pear fruits are shown in this article. According obtained results significant differences among cultivars are confirmed and they could be divided in four groups. Most susceptibly cultivars were Santa Marija. Second group includes Williams, Morettini, Carmen, Hardenpont. As most resistant shown to be Magness, Turandot and two local varietyies Karamanka, as well as another unknown local cultivar. ...

  11. Absence of lysogeny in wild populations of Erwinia amylovora and Pantoea agglomerans.

    Science.gov (United States)

    Roach, Dwayne R; Sjaarda, David R; Sjaarda, Calvin P; Ayala, Carlos Juarez; Howcroft, Brittany; Castle, Alan J; Svircev, Antonet M

    2015-05-01

    Lytic bacteriophages are in development as biological control agents for the prevention of fire blight disease caused by Erwinia amylovora. Temperate phages should be excluded as biologicals since lysogeny produces the dual risks of host resistance to phage attack and the transduction of virulence determinants between bacteria. The extent of lysogeny was estimated in wild populations of E. amylovora and Pantoea agglomerans with real-time polymerase chain reaction primers developed to detect E. amylovora phages belonging to the Myoviridae and Podoviridae families. Pantoea agglomerans, an orchard epiphyte, is easily infected by Erwinia spp. phages, and it serves as a carrier in the development of the phage-mediated biological control agent. Screening of 161 E. amylovora isolates from 16 distinct geographical areas in North America, Europe, North Africa and New Zealand and 82 P. agglomerans isolates from southern Ontario, Canada showed that none possessed prophage. Unstable phage resistant clones or lysogens were produced under laboratory conditions. Additionally, a stable lysogen was recovered from infection of bacterial isolate Ea110R with Podoviridae phage ΦEa35-20. These laboratory observations suggested that while lysogeny is possible in E. amylovora, it is rare or absent in natural populations, and there is a minimal risk associated with lysogenic conversion and transduction by Erwinia spp. phages.

  12. Fe2+ chelator proferrorosamine A: a gene cluster of Erwinia rhapontici P45 involved in its synthesis and its impact on growth of Erwinia amylovora CFBP1430.

    Science.gov (United States)

    Born, Yannick; Remus-Emsermann, Mitja N P; Bieri, Marco; Kamber, Tim; Piel, Jörn; Pelludat, Cosima

    2016-02-01

    Proferrorosamine A (proFRA) is an iron (Fe2+) chelator produced by the opportunistic plant pathogen Erwinia rhapontici P45. To identify genes involved in proFRA synthesis, transposon mutagenesis was performed. The identified 9.3 kb gene cluster, comprising seven genes, designated rosA-rosG, encodes proteins that are involved in proFRA synthesis. Based on gene homologies, a biosynthetic pathway model for proFRA is proposed. To obtain a better understanding of the effect of proFRA on non-proFRA producing bacteria, E. rhapontici P45 was co-cultured with Erwinia amylovora CFBP1430, a fire-blight-causing plant pathogen. E. rhapontici P45, but not corresponding proFRA-negative mutants, led to a pink coloration of E. amylovora CFBP1430 colonies on King's B agar, indicating accumulation of the proFRA-iron complex ferrorosamine, and growth inhibition in vitro. By saturating proFRA-containing extracts with Fe2+, the inhibitory effect was neutralized, suggesting that the iron-chelating capability of proFRA is responsible for the growth inhibition of E. amylovora CFBP1430.

  13. Low genetic diversity among pathogenic strains of Erwinia psidii from Brazil Baixa diversidade genética entre estirpes patogênicas de Erwinia psidii no Brasil

    Directory of Open Access Journals (Sweden)

    Ana C. O. Teixeira

    2009-09-01

    Full Text Available Erwinia psidii causes bacterial disease of guava in Brazil. Phenotypic and molecular characterization through rep-PCR fingerprinting of 42 strains from different geographical regions showed that E. psidii populations in Brazil have a low level of genetic diversity and suggest that contaminated plant material is the main source for pathogen dissemination in the country.Erwinia psidii é o agente causal da seca-dos-ponteiros ou bacteriose da goiabeira no Brasil. A caracterização fenotípica e molecular através de rep-PCR de 42 estirpes patogênicas de diferentes regiões mostrou que as populações de E. psidii no Brasil têm um baixo nível de diversidade genética e sugere que material de propagação infectado é a principal fonte de disseminação do patógeno para novas áreas no país.

  14. Erwinia amylovora expresses fast and simultaneously hrp/dsp virulence genes during flower infection on apple trees.

    Directory of Open Access Journals (Sweden)

    Doris Pester

    Full Text Available BACKGROUND: Pathogen entry through host blossoms is the predominant infection pathway of the gram-negative bacterium Erwinia amylovora leading to manifestation of the disease fire blight. Like in other economically important plant pathogens, E. amylovora pathogenicity depends on a type III secretion system encoded by hrp genes. However, timing and transcriptional order of hrp gene expression during flower infections are unknown. METHODOLOGY/PRINCIPAL FINDINGS: Using quantitative real-time PCR analyses, we addressed the questions of how fast, strong and uniform key hrp virulence genes and the effector dspA/E are expressed when bacteria enter flowers provided with the full defense mechanism of the apple plant. In non-invasive bacterial inoculations of apple flowers still attached to the tree, E. amylovora activated expression of key type III secretion genes in a narrow time window, mounting in a single expression peak of all investigated hrp/dspA/E genes around 24-48 h post inoculation (hpi. This single expression peak coincided with a single depression in the plant PR-1 expression at 24 hpi indicating transient manipulation of the salicylic acid pathway as one target of E. amylovora type III effectors. Expression of hrp/dspA/E genes was highly correlated to expression of the regulator hrpL and relative transcript abundances followed the ratio: hrpA>hrpN>hrpL>dspA/E. Acidic conditions (pH 4 in flower infections led to reduced virulence/effector gene expression without the typical expression peak observed under natural conditions (pH 7. CONCLUSION/SIGNIFICANCE: The simultaneous expression of hrpL, hrpA, hrpN, and the effector dspA/E during early floral infection indicates that speed and immediate effector transmission is important for successful plant invasion. When this delicate balance is disturbed, e.g., by acidic pH during infection, virulence gene expression is reduced, thus partly explaining the efficacy of acidification in fire blight

  15. Potential Distribution of Alien Invasive Species and Risk Assessment: a Case Study of Erwinia amylovora in China

    Institute of Scientific and Technical Information of China (English)

    CHEN Chen; CHEN Juan; HU Bai-shi; JIANG Ying-hua; LIU Feng-quan

    2007-01-01

    Alien invasive species represent a severe risk to biodiversity and economy, as in the case of fire blight (Erwinia amylovora), a bacterial disease that originated in North America, which may be released into new locations by means of fruit trade. On the basis of the knowledge of Erwinia amylovora's biophysical characteristics and environmental data, the geographic information system (GIS) has been applied to determine areas where Erwinia amylovora can potentially invade China. Temperature and precipitation, during the blossoming period, are considered to be two critical factors affecting the Erwinia amylovora's suitable climatic zones. This spatial modeling approach was validated from a case study in Europe, where the occurrence of Erwinia amylovora has been proven. The model prediction agreed with the occurrence of the bacteria recorded in Europe, and the same procedure has been applied to produce a potential establishment area in China's two preferential apple cultivation regions, Bohai Bay region and Huangtu Altiplano region. It has been found that areas belonging to the high-risk category are more or less the main apple producing areas, accounting for their great economic importance in China. This methodology provides an initial baseline for assessment, prevention, and management of alien species that may become invasive under certain environmental conditions. In addition, this modeling approach provides a tool for policy makers to use, in making decisions on management practices where alien species are involved.

  16. The olive fly endosymbiont, "Candidatus Erwinia dacicola," switches from an intracellular existence to an extracellular existence during host insect development.

    Science.gov (United States)

    Estes, Anne M; Hearn, David J; Bronstein, Judith L; Pierson, Elizabeth A

    2009-11-01

    As polyphagous, holometabolous insects, tephritid fruit flies (Diptera: Tephritidae) provide a unique habitat for endosymbiotic bacteria, especially those microbes associated with the digestive system. Here we examine the endosymbiont of the olive fly [Bactrocera oleae (Rossi) (Diptera: Tephritidae)], a tephritid of great economic importance. "Candidatus Erwinia dacicola" was found in the digestive systems of all life stages of wild olive flies from the southwestern United States. PCR and microscopy demonstrated that "Ca. Erwinia dacicola" resided intracellularly in the gastric ceca of the larval midgut but extracellularly in the lumen of the foregut and ovipositor diverticulum of adult flies. "Ca. Erwinia dacicola" is one of the few nonpathogenic endosymbionts that transitions between intracellular and extracellular lifestyles during specific stages of the host's life cycle. Another unique feature of the olive fly endosymbiont is that unlike obligate endosymbionts of monophagous insects, "Ca. Erwinia dacicola" has a G+C nucleotide composition similar to those of closely related plant-pathogenic and free-living bacteria. These two characteristics of "Ca. Erwinia dacicola," the ability to transition between intracellular and extracellular lifestyles and a G+C nucleotide composition similar to those of free-living relatives, may facilitate survival in a changing environment during the development of a polyphagous, holometabolous host. We propose that insect-bacterial symbioses should be classified based on the environment that the host provides to the endosymbiont (the endosymbiont environment).

  17. The Analysis of Saccharides in Chrysanthemi Flos by Capillary Electrophoresis-Amperometric Detection%菊花中糖类组分的毛细管电泳——安培检测研究

    Institute of Scientific and Technical Information of China (English)

    朱金坤; 姚孝林; 郑胜彪; 唐婧

    2012-01-01

    本文采用毛细管区带电泳-安培检测法(CZE-AD),研究了四类菊花中的常见单糖及蔗糖的分离、检测方法。在选定的实验条件下,可在20分钟内实现对葡萄糖、果糖、半乳糖、蔗糖的有效分离;待测糖类在铜电极上具有良好的电流响应、检测灵敏度(LOD:~1.0×10-6mol/L)及适宜的线性范围(5.0×10-6~5.0×10-4mol/L)。实验对滁菊、杭黄菊、贡菊、亳菊中的糖类组分进行了CZE-AD分离检测,方法具有良好的重现性及应用前景。检测结果表明,四类菊花中常见糖类组分的含量差异较大,可为相关植物资源的开发利用提供参考;检测方法简单、快速、有效,可用于菊花中常见糖类成分的测定及质量控制。%In this paper,the determination of normal saccharides in four kinds of Chrysanthemi Flos was investigated with capillary zone electrophoresis-amperometric detection method(CZE-AD).Under the optimized conditions,glucose,fructose,galactose and sucrose could be separated in 20 min.These analyzed saccharides gained excellent current response,detection limits(LOD): ~1.0×10-6mol/L and adequate linear ranges(5.0×10-6~5.0×10-4mol/L).The saccharides in four kinds of Chrysanthemi Flos(Chu-Ju,Hanghuang-Ju,Gong-Ju,Bo-Ju) were analyzed,and the results indicated good reproducibility and applicability of this proposed CZE-AD method.It could be found that the concerns of saccharides varied from one kind of Chrysanthemi Flos to another,and the detection data would provide reference to exploit the corresponding plant resources efficiently.This proposed detection method was simple,quick and effective with potential application in the analysis of saccharides for quality control of Chrysanthemi Flos.

  18. EFEKTIFITAS DAYA HAMBAT BAKTERI Streptomyces sp TERHADAP Erwinia sp PENYEBAB PENYAKIT BUSUK REBAH PADA TANAMAN LIDAH BUAYA (Aloe barbadensis Mill

    Directory of Open Access Journals (Sweden)

    SARMILA TASNIM

    2013-05-01

    Full Text Available Streptomyces sp was conducted from December 2010 - June 2011 at the Laboratoryof Microbiology, Biology Department, Math and Science Faculty, UdayanaUniversity Bukit Jimbaran-Bali. Implementation stages of the research consisted ofisolation and testing of the antibiotic activity Streptomyces sp to inhibit growthbacterial pathogens Erwinia sp as a cause of disease in plants fallen foul (Soft rot ofAloe barbadensis Mill.The results of this study have eight isolates of Streptomyces spwith macroscopic and microscopic characters are varied. Furthermore, all isolateswere obtained and then tested against antibiotic activity to inhibit growth the bacteriaErwinia sp. Test results obtained by Streptomyces sp that has the most effective ininhibiting the ability of the bacteria Erwinia sp isolates are Streptomyces sp2for (45%.

  19. Genetic differences between blight-causing Erwinia species with differing host specificities, identified by suppression subtractive hybridization.

    Science.gov (United States)

    Triplett, Lindsay R; Zhao, Youfu; Sundin, George W

    2006-11-01

    PCR-based subtractive hybridization was used to isolate sequences from Erwinia amylovora strain Ea110, which is pathogenic on apples and pears, that were not present in three closely related strains with differing host specificities: E. amylovora MR1, which is pathogenic only on Rubus spp.; Erwinia pyrifoliae Ep1/96, the causal agent of shoot blight of Asian pears; and Erwinia sp. strain Ejp556, the causal agent of bacterial shoot blight of pear in Japan. In total, six subtractive libraries were constructed and analyzed. Recovered sequences included type III secretion components, hypothetical membrane proteins, and ATP-binding proteins. In addition, we identified an Ea110-specific sequence with homology to a type III secretion apparatus component of the insect endosymbiont Sodalis glossinidius, as well as an Ep1/96-specific sequence with homology to the Yersinia pestis effector protein tyrosine phosphatase YopH.

  20. High performance liquid chromatography time of flight electrospray ionization mass spectrometry for quantification of sesquiterpenes in Chrysanthemi indici Flos active extract

    Science.gov (United States)

    Fu, Ling; Wang, Pan; Sun, Yiqun; Wang, Yangyang; Zhao, Jing; Ye, Yuting; Zhang, Yanbin; Bi, Yuefeng

    2015-01-01

    Background: Chrysanthemi indici Flos, a traditional herbal medicine is used to clearing heat–toxicity, removing the liver fire, and improving eyesight. In our preliminary work, an active extract of CTC in C. An indici Flos with anti-hepatitis B virus and liver protective activity was found by HepG2.2.1.5 test and experiment of protein synthesis in mice's injured liver. In this work, we aimed to study the active faction CTC further by qualitative and quantitative analysis method. Materials and Methods: High performance liquid chromatography time of flight electrospray ionization mass spectrometry (HPLC TOF ESI-MS) analysis method of the CTC was established. Cumambrin A and angeloylcumambrin B in CTC were analyzed by high performance liquid chromatography-ultraviolet-evaporative light scattering detector (HPLC-UV-ELSD) analysis methods. A binary gradient elution program was conducted for chromatographic separation with acetonitrile (A) and ultrapure water (B) as follows: 0–10 min, 42–46% A; 10–20 min, 46–55% A; 20–25 min, 55–60% A; and 25–35 min, 60–75% A. The column temperature and UV wavelength were set at 30°C and 205 nm. Result: Ten constituents including (3R, 5R, 6S, 7S, 10R)-7-(2-hydroxy-2-propyl)-10-methyl-4-methyleneperhy, dronaphthal-ene-3, 5, 6-triol acetone solvate; (+)-edusmance-4, (14)-ene-11, 13-diol; linarin; luteolin; apigenin; tricin; 5, 3’,4’- trimethyl-6,7-dimethoxy flavones; cumambrin A; acacetin; and angeloylcumambrin B in CTC were identified by HPLC TOF ESI-MS. The contents of sesquiterpenes in CTC were decreased by storing years. Conclusions: The results showed that both UV and ELSD methods were feasible, accurate, and the determination results were in good consistency. The contents of two sesquiterpenes decreased with storing years. Two sesquiterpenes could be used as quality control for C. indici flos CTC. PMID:26600718

  1. Relaciones serológicas entre aislamientos bacterianos de los géneros Erwinia, Pectobacterium Y Pantoea

    OpenAIRE

    Yonis Hernández; Gustavo Trujillo

    2004-01-01

    Este estudio fue realizado con el propósito de conocer las relaciones serológicas entre aislamientos pertenecientes a diferentes generos y especies de bacterias. Se utilizaron ocho aislamientos, tres de Pectobacterium chrysanthemi obtenidos de maíz (Zea mays), papa (Solanum tuberosum) y batata (Ipomoea batatas); tres de P. carotovora subsp. carotovora, provenientes uno de cafecito de jardín (Aglaonema commutatum ´María‘) y dos de tomate (Lycopersicon esculentum), con diferencias en el comport...

  2. Erwinia asparaginase achieves therapeutic activity after pegaspargase allergy: a report from the Children's Oncology Group.

    Science.gov (United States)

    Salzer, Wanda L; Asselin, Barbara; Supko, Jeffrey G; Devidas, Meenakshi; Kaiser, Nicole A; Plourde, Paul; Winick, Naomi J; Reaman, Gregory H; Raetz, Elizabeth; Carroll, William L; Hunger, Stephen P

    2013-07-25

    AALL07P2 evaluated whether substitution of Erwinia asparaginase 25000 IU/m(2) for 6 doses given intramuscularly Monday/Wednesday/Friday (M/W/F) to children and young adults with acute lymphoblastic leukemia and clinical allergy to pegaspargase would provide a 48-hour nadir serum asparaginase activity (NSAA) ≥ 0.10 IU/mL. AALL07P2 enrolled 55 eligible/evaluable patients. NSAA ≥ 0.1 IU/mL was achieved in 38 of 41 patients (92.7%) with acceptable samples 48 hours and in 38 of 43 patients (88.4%) 72 hours after dosing during course 1. Among samples obtained during all courses, 95.8% (252 of 263) of 48-hour samples and 84.5% (125 of 148) of 72-hour samples had NSAA ≥ 0.10-IU/mL. Pharmacokinetic parameters were estimated by fitting the serum asparaginase activity-time course for all 6 doses given during course 1 to a 1-compartment open model with first order absorption. Erwinia asparaginase administered with this schedule achieved therapeutic NSAA at both 48 and 72 hours and was well tolerated with no reports of hemorrhage, thrombosis, or death, and few cases of grade 2 to 3 allergic reaction (n = 6), grade 1 to 3 hyperglycemia (n = 6), or grade 1 pancreatitis (n = 1). Following allergy to pegaspargase, Erwinia asparaginase 25000 IU/m(2) × 6 intramuscularly M/W/F can be substituted for a single dose of pegaspargase.

  3. Biology of the fire blight pathogen Erwinia amylovora in oligotrophic environments: survival responses and virulence

    OpenAIRE

    Delgado Santander, Ricardo

    2016-01-01

    Erwinia amylovora es una bacteria fitopatógena de la familia Enterobacteriaceae, responsable del fuego bacteriano de las rosáceas. Los efectos destructivos de este patógeno sobre frutos, flores y prácticamente todos los órganos de las plantas hospedadoras afectadas constituyen una amenaza importante para la producción de pera y manzana, y suponen graves pérdidas económicas anuales en todo el mundo. E. amylovora está clasificada como un organismo de cuarentena en la Unión Europea y en otros pa...

  4. Erwinia amylovora – the Causal Agent of Root Collar Necrosis of Apple Trees

    OpenAIRE

    Veljko Gavrilović; Svetlana Milijašević; Biljana Todorović; Svetlana Živković; Nenad Trkulja

    2008-01-01

    A large-scale outbreak of fire blight symptoms caused by Erwinia amylovora was recorded in pome fruit trees during 2007. In addition to fruit necrosis and shoot blight as the typical disease symptoms, dark purple necrosis was observed in the root collar area girdling the trunk just above the ground and thus withering the whole apple tree. Since similar symptoms on apple trees could be caused by E. amylovora or one of several phytopathogenic fungi of the genera Phomopsis and Phytophthora, an i...

  5. Same ammo, different weapons: enzymatic extracts from two apple genotypes with contrasted susceptibilities to fire blight (Erwinia amylovora) differentially convert phloridzin and phloretin in vitro.

    Science.gov (United States)

    Gaucher, Matthieu; Dugé de Bernonville, Thomas; Guyot, Sylvain; Dat, James F; Brisset, Marie-Noëlle

    2013-11-01

    The necrogenic bacterium Erwinia amylovora responsible for the fire blight disease causes cell death in apple tissues to enrich intercellular spaces with nutrients. Apple leaves contain large amounts of dihydrochalcones (DHCs), including phloridzin and its aglycone phloretin. Previous work showed an important decrease in the constitutive DHCs stock in infected leaves, probably caused by transformation reactions during the infection process. At least two flavonoid transformation pathways have been described so far: deglucosylation and oxidation. The aim of the present study was to determine whether DHCs are differentially converted in two apple genotypes displaying contrasted susceptibilities to the disease. Different analyses were performed: i) enzymatic activity assays in infected leaves, ii) identification/quantification of end-products obtained after in vitro enzymatic reactions with DHCs, iii) evaluation of the bactericidal activity of end-products. The results of the enzymatic assays showed that deglucosylation was dominant over oxidation in the susceptible genotype MM106 while the opposite was observed in the resistant genotype Evereste. These data were confirmed by LC-UV/Vis-MS analysis of in vitro reaction mixtures, especially because higher levels of o-quinoid oxidation products of phloretin were measured by using the enzymatic extracts of Evereste infected leaves. Their presence correlated well with a strong bactericidal activity of the reaction mixtures. Thus, our results suggest that a differential transformation of DHCs occur in apple genotypes with a potential involvement in the establishment of the susceptibility or the resistance to fire blight, through the release of glucose or of highly bactericidal compounds respectively.

  6. Surface survival and internalization of salmonella through natural cracks on developing cantaloupe fruits, alone or in the presence of the melon wilt pathogen Erwinia tracheiphila.

    Directory of Open Access Journals (Sweden)

    Dhiraj Gautam

    Full Text Available Outbreaks of foodborne illness attributed to the consumption of Salmonella-tainted cantaloupe have occurred repeatedly, but understanding of the ecology of Salmonella on cantaloupe fruit surfaces is limited. We investigated the interactions between Salmonella enterica Poona, the plant pathogenic bacterium Erwinia tracheiphila, and cantaloupe fruit. Fruit surfaces were inoculated at the natural cracking stage by spreading S. enterica and E. tracheiphila, 20 µl at 107 cfu/ml, independently or together, over a 2×2 cm rind area containing a crack. Microbial and microscopic analyses were performed at 0, 9 and 24 days post inoculation (DPI. Even at 24 DPI (fruit maturity S. enterica was detected on 14% and 40% of the fruit inoculated with S. enterica alone and the two-pathogen mixture, respectively. However, the population of S. enterica declined gradually after initial inoculation. E. tracheiphila, inoculated alone or together with Salmonella, caused watersoaked lesions on cantaloupe fruit; but we could not conclude in this study that S. enterica survival on the fruit surface was enhanced by the presence of those lesions. Of fruit inoculated with E. tracheiphila alone and sampled at 24 DPI, 61% had watersoaked lesions on the surface. In nearly half of those symptomatic fruits the watersoaking extended into the sub-rind mesocarp, and E. tracheiphila was recovered from that tissue in 50% of the symptomatic fruit. In this work, E. tracheiphila internalized through natural cracks on developing fruits. S. enterica was never detected in the fruit interior (ca. 2-3 mm below rind surface under the limited conditions of our experiments, but the possibility that it, or other human pathogens that contaminate fresh produce, might also do so should be investigated under a wider range of conditions and produce types.

  7. Evidence of two lineages of the symbiont 'Candidatus Erwinia dacicola' in Italian populations of Bactrocera oleae (Rossi) based on 16S rRNA gene sequences.

    Science.gov (United States)

    Savio, Claudia; Mazzon, Luca; Martinez-Sañudo, Isabel; Simonato, Mauro; Squartini, Andrea; Girolami, Vincenzo

    2012-01-01

    The close association between the olive fly Bactrocera oleae (Rossi) (Diptera: Tephritidae) and bacteria has been known for more than a century. Recently, the presence of a host-specific, hereditary, unculturable symbiotic bacterium, designated 'Candidatus Erwinia dacicola', has been described inside the cephalic organ of the fly, called the oesophageal bulb. In the present study, the 16S rRNA gene sequence variability of 'Ca. E. dacicola' was examined within and between 26 Italian olive fly populations sampled across areas where olive trees occur in the wild and areas where cultivated olive trees have been introduced through history. The bacterial contents of the oesophageal bulbs of 314 olive flies were analysed and a minimum of 781 bp of the 16S rRNA gene was sequenced. The corresponding host fly genotype was assessed by sequencing a 776 bp portion of the mitochondrial genome. Two 'Ca. E. dacicola' haplotypes were found (htA and htB), one being slightly more prevalent than the other (57%). The two haplotypes did not co-exist in the same individuals, as confirmed by cloning. Interestingly, the olive fly populations of the two main Italian islands, Sicily and Sardinia, appeared to be represented exclusively by the htB and htA haplotypes, respectively, while peninsular populations showed both bacterial haplotypes in different proportions. No significant correlation emerged between the two symbiont haplotypes and the 16 host fly haplotypes observed, suggesting evidence for a mixed model of vertical and horizontal transmission of the symbiont during the fly life cycle.

  8. Surface survival and internalization of salmonella through natural cracks on developing cantaloupe fruits, alone or in the presence of the melon wilt pathogen Erwinia tracheiphila.

    Science.gov (United States)

    Gautam, Dhiraj; Dobhal, Shefali; Payton, Mark E; Fletcher, Jacqueline; Ma, Li Maria

    2014-01-01

    Outbreaks of foodborne illness attributed to the consumption of Salmonella-tainted cantaloupe have occurred repeatedly, but understanding of the ecology of Salmonella on cantaloupe fruit surfaces is limited. We investigated the interactions between Salmonella enterica Poona, the plant pathogenic bacterium Erwinia tracheiphila, and cantaloupe fruit. Fruit surfaces were inoculated at the natural cracking stage by spreading S. enterica and E. tracheiphila, 20 µl at 107 cfu/ml, independently or together, over a 2×2 cm rind area containing a crack. Microbial and microscopic analyses were performed at 0, 9 and 24 days post inoculation (DPI). Even at 24 DPI (fruit maturity) S. enterica was detected on 14% and 40% of the fruit inoculated with S. enterica alone and the two-pathogen mixture, respectively. However, the population of S. enterica declined gradually after initial inoculation. E. tracheiphila, inoculated alone or together with Salmonella, caused watersoaked lesions on cantaloupe fruit; but we could not conclude in this study that S. enterica survival on the fruit surface was enhanced by the presence of those lesions. Of fruit inoculated with E. tracheiphila alone and sampled at 24 DPI, 61% had watersoaked lesions on the surface. In nearly half of those symptomatic fruits the watersoaking extended into the sub-rind mesocarp, and E. tracheiphila was recovered from that tissue in 50% of the symptomatic fruit. In this work, E. tracheiphila internalized through natural cracks on developing fruits. S. enterica was never detected in the fruit interior (ca. 2-3 mm below rind surface) under the limited conditions of our experiments, but the possibility that it, or other human pathogens that contaminate fresh produce, might also do so should be investigated under a wider range of conditions and produce types.

  9. Bacterierot in Sedum hoe te handelen

    NARCIS (Netherlands)

    Helm, van der F.P.M.

    2009-01-01

    In sedum kan een bacterieziekte voorkomen die zich bij nat weer zeer snel door het gewas verpreidt. De vermoedelijke veroorzaker is Erwinia chrysanthemi (Dickeya). Deze bacterie zorgt in hyacint voor het gevreesde agressieve witsnot en in dahlia voor bacterieverwelkingsziekte

  10. Identification of genes differentially expressed during interaction of resistant and susceptible apple cultivars (Malus x domestica) with Erwinia amylovora

    Science.gov (United States)

    The necrogenic enterobacterium, Erwinia amylovora is the causal agent of the fire blight (FB) disease in many Rosaceae species, including apple and pear. During the infection process, the bacteria induce an oxidative stress response with kinetics similar to those induced in an incompatible bacteria-...

  11. Apple (Malus x domestica) transcriptome in response to the compatible pathogen Erwinia amylovora and the incompatible pathogen Pseudomonas syringae

    Science.gov (United States)

    Infiltration of Erwinia amylovora (Ea) into host leaves induces an oxidative burst similar to that observed during incompatible reactions associated with Hypersensitive Response (HR). However, the subsequent progressive development of necrosis in apple and other hosts is unlike an incompatible reac...

  12. TRANSGENIC EXPRESSION OF THE ERWINIA AMYLOVORA (FIRE BLIGHT) EFFECTOR PROTEIN EOP1 SUPRESSES HOST BASAL DEFENSE MECHANISMS IN MALUS (APPLE)

    Science.gov (United States)

    Erwinia amylovora (Ea) is the causative agent of fire blight, a devastating disease of apple and pear. Like many other plant and animal bacterial pathogens Ea utilizes a type three secretion system (TTSS) to deliver effector proteins into plant host cells. Once inside the host cell, effector protei...

  13. Prevalence of Candidatus Erwinia dacicola in wild and laboratory olive fruit fly populations and across developmental stages.

    Science.gov (United States)

    Estes, Anne M; Hearn, David J; Burrack, Hannah J; Rempoulakis, Polychronis; Pierson, Elizabeth A

    2012-04-01

    The microbiome of the olive fruit fly, Bactrocera oleae (Gmelin), a worldwide pest of olives (Olea europaea L.), has been examined for >100 yr as part of efforts to identify bacteria that are plant pathogens vectored by the fly or are beneficial endosymbionts essential for the fly's survival and thus targets for possible biological control. Because tephritid fruit flies feed on free-living bacteria in their environment, distinguishing between the transient, acquired bacteria of their diet and persistent, resident bacteria that are vertically transmitted endosymbionts is difficult. Several culture-dependent and -independent studies have identified a diversity of species in the olive fruit fly microbiome, but they have not distinguished the roles of the microbes. Candidatus Erwinia dacicola, has been proposed to be a coevolved endosymbiont of the olive fruit fly; however, this was based on limited samples from two Italian populations. Our study shows that C. Erwinia dacicola was present in all New and Old World populations and in the majority of individuals of all life stages sampled in 2 yr. Olive fruit flies reared on olives in the laboratory had frequencies of C. Erwinia dacicola similar to that of wild populations; however, flies reared on artificial diets containing antibiotics in the laboratory rarely had the endosymbiont. The relative abundance of C. Erwinia dacicola varied across development stages, being most abundant in ovipositing females and larvae. This uniform presence of C. Erwini dacicola suggests that it is a persistent, resident endosymbiont of the olive fruit fly.

  14. Conductrimetric detection of Pseudomonas syringae pathovar pisi in pea seeds and soft rot Erwinia spp. on potato tubers.

    NARCIS (Netherlands)

    Fraaije, B.A.

    1996-01-01

    Pea bacterial blight and potato blackleg are diseases caused by Pseudomonas syringae pv. pisi ( Psp ) and soft rot Erwinia spp., respectively. The primary source of inoculum for these bacteria is contaminated plant propagation material, i.e. pea seeds and potato tubers. One of the best ways to contr

  15. Genetics of biosynthesis and structure of the capsular exopolysaccharide from the Asian pear pathogen Erwinia pyrifoliae.

    Science.gov (United States)

    Kim, Won-Sik; Schollmeyer, Martin; Nimtz, Manfred; Wray, Victor; Geider, Klaus

    2002-12-01

    Erwinia pyrifoliae is a novel bacterial pathogen, which causes Asian pear blight and is related to Erwinia amylovora, the causative agent of fire blight. E. pyrifoliae produces exopolysaccharide (EPS) related to amylovoran in its sugar composition and sugar linkages. This was shown by degradation of the EPS with a viral depolymerase, and by methylation analysis and ESI/MS. The structure of the repeating units was confirmed by (1)H-NMR spectra. The EPS of E. pyrifoliae carried side chains, which were mainly terminated by acetyl and pyruvyl residues as found previously for amylovoran. On the other hand, a second side chain with glucose found for up to 65% of the repeating units of amylovoran was completely absent. The nucleotide sequences of five genes of the cps cluster of E. pyrifoliae encoding proteins for EPS synthesis were characterized and displayed a high homology with the corresponding ams genes. Similar functions of the gene products are assumed. As for ams mutants of E. amylovora, a cpsB mutant of E. pyrifoliae did not synthesize EPS and did not produce ooze on slices of immature pears or symptoms on pear seedlings. The cps mutant was complemented for EPS synthesis and virulence on pear slices with a gene cluster of E. amylovora that included amsB.

  16. Molecular detection of Erwinia psidii in guava plants under greenhouse and field conditions

    Directory of Open Access Journals (Sweden)

    Claudênia Ferreira da Silva

    2016-09-01

    Full Text Available ABSTRACT: Erwinia psidii causes bacterial blight of guava ( Psidium guajava , an important disease of this crop in Brazil. The pathogen affects branches and twigs of guava trees, reducing yield significantly. Bacterial dissemination often occurs through contaminated but asymptomatic propagating plant material. The objectives of this research were to evaluate the use of BIO-PCR and conventional PCR to detect E. psidii in inoculated guava plants grown in a greenhouse and in symptomatic and asymptomatic trees from guava orchards. Erwinia psidii strain IBSBF 1576 was inoculated (107CFU mL-1 into young guava shoots and plant tissue was analysed at 0, 5, 10, and 15 days after inoculation. Symptoms were observed after 5 days and all inoculated shoots were PCR positive at all times, by both BIO-PCR and conventional PCR. Under natural infection conditions, 40 samples were tested by BIO-PCR from each of three guava orchards, 20 showing symptoms and 20 asymptomatic. PCR was positive for 58 out of 60 symptomatic samples (96.7% and for 6.7% of asymptomatic samples, showing that the method can be used to detect the pathogen at early stages of infection. This PCR method may be used as a diagnostic tool to assess bacterial survival, dissemination and disease outbreaks.

  17. Preliminary results on the ability of pentatomidae to transfer fire blight Erwinia amylovora under controlled conditions.

    Science.gov (United States)

    Peusens, G; Schoofs, H; Deckers, T; Belien, T

    2013-01-01

    With their piercing-sucking mouthparts stink bugs (Heteroptera: Pentatomidae), a major pest in especially organic orchards, create wounds in fruit of pear trees. As Erwinia amylovora (Burrill, Winslow et al.), a wide spread bacterial disease affecting many rosaceous plants including pome fruit trees and hawthorn, enters through openings in flowers, leaves, shoots and fruit, feeding punctures caused by these bugs might be inoculated with Erwinia bacteria. In order to investigate the ability of the bugs Pentotoma rufipes L. and Polomena prasina L. to transmit fire blight, insects were caught in an organically managed orchard without fire blight, brought into contact with artificially inoculated immature pear fruit/slices and transferred to healthy, mechanically wounded pear fruit/slices. After an incubation period potential transmission of bacteria was examined by evaluation of symptom expression (necrosis, ooze production). To assess the presence of bacteria on the exoskeleton of the tested bugs, all bugs were forced to walk on a semiselective nutrient agar medium. In another experiment the viability of Ea on the exoskeleton was tested -after previous contact with ooze- through washing and plating of the wash water. All experiments were conducted under optimal climatological conditions and according to quarantine standards. Results demonstrated the ability of stink bugs to transfer E. amylovora to fruit and the viability of bacteria on stink bugs externally - both under lab conditions.

  18. A prospective study on drug monitoring of PEGasparaginase and Erwinia asparaginase and asparaginase antibodies in pediatric acute lymphoblastic leukemia.

    Science.gov (United States)

    Tong, Wing H; Pieters, Rob; Kaspers, Gertjan J L; te Loo, D Maroeska W M; Bierings, Marc B; van den Bos, Cor; Kollen, Wouter J W; Hop, Wim C J; Lanvers-Kaminsky, Claudia; Relling, Mary V; Tissing, Wim J E; van der Sluis, Inge M

    2014-03-27

    This study prospectively analyzed the efficacy of very prolonged courses of pegylated Escherichia coli asparaginase (PEGasparaginase) and Erwinia asparaginase in pediatric acute lymphoblastic leukemia (ALL) patients. Patients received 15 PEGasparaginase infusions (2500 IU/m(2) every 2 weeks) in intensification after receiving native E coli asparaginase in induction. In case of allergy to or silent inactivation of PEGasparaginase, Erwinia asparaginase (20 000 IU/m(2) 2-3 times weekly) was given. Eighty-nine patients were enrolled in the PEGasparaginase study. Twenty (22%) of the PEGasparaginase-treated patients developed an allergy; 7 (8%) showed silent inactivation. The PEGasparaginase level was 0 in all allergic patients (grade 1-4). Patients without hypersensitivity to PEGasparaginase had serum mean trough levels of 899 U/L. Fifty-nine patients were included in the Erwinia asparaginase study; 2 (3%) developed an allergy and none silent inactivation. Ninety-six percent had at least 1 trough level ≥100 U/L. The serum asparagine level was not always completely depleted with Erwinia asparaginase in contrast to PEGasparaginase. The presence of asparaginase antibodies was related to allergies and silent inactivation, but with low specificity (64%). Use of native E coli asparaginase in induction leads to high hypersensitivity rates to PEGasparaginase in intensification. Therefore, PEGasparaginase should be used upfront in induction, and we suggest that the dose could be lowered. Switching to Erwinia asparaginase leads to effective asparaginase levels in most patients. Therapeutic drug monitoring has been added to our ALL-11 protocol to individualize asparaginase therapy.

  19. Dickeya solani sp. nov., a pectinolytic plant pathogenic bacterium isolated from potato (Solanum tuberosum)

    NARCIS (Netherlands)

    Wolf, van der J.M.; Nijhuis, E.H.; Kowalewska, M.J.; Saddler, G.S.; Parkinson, N.; Elphinstone, J.G.; Pritchard, L.; Toth, I.K.; Lojkowska, E.; Potrykus, M.; Waleron, M.; Vos, de P.; Cleenwerck, I.; Pirhonen, M.; Garlant, L.; Hélias, V.; Pothier, J.F.; Pflüger, V.; Duffy, B.; Tsror, L.; Manulis, S.

    2014-01-01

    Pectinolytic bacteria were recently isolated from diseased potato plants exhibiting blackleg and slow wilt symptoms found in a number of European countries and Israel. These Gram-negative, motile, rods were identified as belonging to the genus Dickeya, previously the Pectobacterium chrysanthemi comp

  20. Phylogenetic position and virulence apparatus of the pear flower necrosis pathogen Erwinia piriflorinigrans CFBP 5888T as assessed by comparative genomics.

    Science.gov (United States)

    Smits, Theo H M; Rezzonico, Fabio; López, María M; Blom, Jochen; Goesmann, Alexander; Frey, Jürg E; Duffy, Brion

    2013-10-01

    Erwinia piriflorinigrans is a necrotrophic pathogen of pear reported from Spain that destroys flowers but does not progress further into the host. We sequenced the complete genome of the type strain CFBP 5888(T) clarifying its phylogenetic position within the genus Erwinia, and indicating a position between its closest relative, the epiphyte Erwinia tasmaniensis and other plant pathogenic Erwinia spp. (i.e., the fire blight pathogen E. amylovora and the Asian pear pathogen E. pyrifoliae). Common features are the type III and type VI secretion systems, amylovoran biosynthesis and desferrioxamine production. The E. piriflorinigrans genome also provided the first evidence for production of the siderophore chrysobactin within the genus Erwinia sensu stricto, which up to now was mostly associated with phytopathogenic, soft-rot Dickeya and Pectobacterium species. Plasmid pEPIR37, reported in this strain, is closely related to small plasmids found in the fire blight pathogen E. amylovora and E. pyrifoliae. The genome of E. piriflorinigrans also gives detailed insights in evolutionary genomics of pathoadapted Erwinia.

  1. Resistance identification of banana varieties to banana bacterial soft rot caused by Dickeya ( Pectobacterium chrysanthemi pv.paradisiaca )%香蕉品种对香蕉细菌性软腐病的抗性鉴定

    Institute of Scientific and Technical Information of China (English)

    蒲小明; 林壁润; 吕顺; 沈会芳; 周佳暖; 周建坤; 韩秀香

    2012-01-01

    Banana bacterial soft rot caused by Dickeya paradisiaca {Pectobacterium chrysanthemi pv. Paradisiaca) was a new devastating invasive disease in China, and using resistant varieties would be an effective method to control the disease. Detached leaf method and pot experiments were used to identify the resistance of 39 banana varieties. The results showed that the levels of resistance of these 39 banana varieties were different, among which 'Huangdi Banana' was highly resistant (HR), 'Red Dwarf' was resistant (R), and 'Aijiao Dundilei' and 'Long-zhou Zhongba' were moderately resistant (MR). Such method could identify resistant varieties effectively, rapidly and accurately, and the resistance results by detached leaf and pot experiments were consistent with those by the field experiments.%由菊欧文氏杆菌香蕉致病变种[Dickeya paradisiaca(原名Pectobacterium chrysanthemi pv.paradisiaca)]引起的香蕉细菌性软腐病是我国新入侵的毁灭性香蕉病害,应用抗病品种将是防治该病害的有效方法.采用离体叶片法和盆栽法相结合的筛选方法对39个香蕉品种进行抗性鉴定,结果表明,供试品种对香蕉细菌性软腐病菌的抗性存在明显差异,其中表现高抗(HR)的品种为‘皇帝蕉’,抗病(R)品种为‘红河矮’,中抗(MR)品种为‘矮脚遁地蕾’和‘龙州中把’.离体和盆栽鉴定结果与品种田间抗病性一致,可有效、快速、准确地筛选出抗病品种.

  2. Cellulose production, activated by cyclic di-GMP through BcsA and BcsZ, is a virulence factor and an essential determinant of the three-dimensional architectures of biofilms formed by Erwinia amylovora Ea1189.

    Science.gov (United States)

    Castiblanco, Luisa F; Sundin, George W

    2016-10-18

    Bacterial biofilms are multicellular aggregates encased in an extracellular matrix mainly composed of exopolysaccharides (EPSs), protein and nucleic acids, which determines the architecture of the biofilm. Erwinia amylovora Ea1189 forms a biofilm inside the xylem of its host, which results in vessel plugging and water transport impairment. The production of the EPSs amylovoran and levan is critical for the formation of a mature biofilm. In addition, cyclic dimeric GMP (c-di-GMP) has been reported to positively regulate amylovoran biosynthesis and biofilm formation in E. amylovora Ea1189. In this study, we demonstrate that cellulose is synthesized by E. amylovora Ea1189 and is a major modulator of the three-dimensional characteristics of biofilms formed by this bacterium, and also contributes to virulence during systemic host invasion. In addition, we demonstrate that the activation of cellulose biosynthesis in E. amylovora is a c-di-GMP-dependent process, through allosteric binding to the cellulose catalytic subunit BcsA. We also report that the endoglucanase BcsZ is a key player in c-di-GMP activation of cellulose biosynthesis. Our results provide evidence of the complex composition of the extracellular matrix produced by E. amylovora and the implications of cellulose biosynthesis in shaping the architecture of the biofilm and in the expression of one of the main virulence phenotypes of this pathogen.

  3. Cloning, purification, crystallization and 1.57 Å resolution X-ray data analysis of AmsI, the tyrosine phosphatase controlling amylovoran biosynthesis in the plant pathogen Erwinia amylovora.

    Science.gov (United States)

    Benini, Stefano; Caputi, Lorenzo; Cianci, Michele

    2014-12-01

    The Gram-negative bacterium Erwinia amylovora is a destructive pathogen of plants belonging to the Rosaceae family. Amongst its pathogenicity factors, E. amylovora produces the exopolysaccharide amylovoran, which contributes to the occlusion of plant vessels, causing wilting of shoots and eventually resulting in plant death. Amylovoran biosynthesis requires the presence of 12 genes (from amsA to amsL) clustered in the ams region of the E. amylovora genome. They mostly encode glycosyl transferases (AmsG, AmsB, AmsD, AmsE, AmsJ and AmsK), proteins involved in amylovoran translocation and assembly (AmsH, AmsL and AmsC), and also a tyrosine kinase (AmsA) and a tyrosine phosphatase (AmsI), which are both involved in the regulation of amylovoran biosynthesis. The low-molecular-weight protein tyrosine phosphatase AmsI was overexpressed as a His6-tagged protein in Escherichia coli, purified and crystallized. X-ray diffraction data were collected to a maximum resolution of 1.57 Å in space group P3121.

  4. Non-conventional possibilities of protection of apple and pear against fire blight (Erwinia amylovora

    Directory of Open Access Journals (Sweden)

    Piotr Sobiczewski

    2013-12-01

    Full Text Available Standard program of plant protection against fire blight consists of use of management practices and chemical control method. Recently a new, non-conventional possibilities based on application of biocontrol agents (two biopreparations have been already introduced into practice: Bliteban A506 (Pseudomonas fluorescens and BlossomBless (Pantoea agglomerans, plant extracts active against Erwinia amylovora (AkseBio containing extracts from Thymbra spicata and Biomit Plussz with extracts from various plant species and microelements and resistance inducers (Regalis, Bion and plant extracts are of great interest. Also plant transformation with resistance genes such as: hrpN (harpin, dpo (EPS depolymerase and lytic protein genes (attacin E, cecropin SB-37, T4 lysozyme is a promising perspective.

  5. Localization of transposon insertions in pathogenicity mutants of Erwinia amylovora and their biochemical characterization.

    Science.gov (United States)

    Bellemann, P; Geider, K

    1992-05-01

    Transposon Tn5, on a mobilizable ColE1 plasmid, on a Ti plasmid derepressed for bacterial transfer, and on the bacteriophage fd genome, was used to construct pathogenicity mutants of the fire blight pathogen Erwinia amylovora. Eleven nonpathogenic mutants were isolated from 1600 independent mutants screened. These mutants were divided into three types: auxotrophs, exopolysaccharide (EPS)-deficient mutants and a mutant of the dsp phenotype. According to their insertion sites the Tn5 mutants were mapped into several classes. Some of the mutants could be complemented with cosmid clones from a genomic library of the parent strain for EPS production on minimal agar. EPS-deficient mutants and the dsp mutant could complement each other to produce virulence symptoms on pear slices.

  6. Structure of amylovoran, the capsular exopolysaccharide from the fire blight pathogen Erwinia amylovora.

    Science.gov (United States)

    Nimtz, M; Mort, A; Domke, T; Wray, V; Zhang, Y; Qiu, F; Coplin, D; Geider, K

    1996-06-01

    The acidic exopolysaccharide (EPS) of Erwinia amylovora, amylovoran, was purified from culture supernatants of bacteria in minimal medium and cleaved chemically either by treatment with trifluoracetic acid or hydrofluoric acid, and enzymatically by digestion with depolymerase from E. amylovora phage phi-Ealh. Structural characterization of the resulting oligosaccharides was performed by a combination of mass spectrometric and NMR [one- and two-dimensional (1D and 2D)] spectroscopic techniques. A branched repeating unit with five monosaccharide residues and various substituents was determined: [sequence: see text] The terminal monosaccharide of the side branch, which bears a 4,6-bound pyruvate residue in the R-configuration, was found to be modified with 2-linked (26%), 3-linked (24%), 2-,3-linked (40%) O-acetyl groups, or these were absent (10%). An additional glucose residue is linked to approximately 10% of the core alpha-galactose of the repeating unit.

  7. Simplification of vacuole structure during plant cell death triggered by culture filtrates of Erwinia carotovora

    Institute of Scientific and Technical Information of China (English)

    Yumi Hirakawa; Toshihisa Nomura; Seiichiro Hasezawa; Takumi Higaki

    2015-01-01

    Vacuoles are suggested to play crucial roles in plant defense-related cel death. During programmed cel death, previous live cel imaging studies have observed vacuoles to become simpler in structure and have implicated this simplification as a prelude to the vacuole’s rupture and consequent lysis of the plasma membrane. Here, we examined dynamics of the vacuole in cel cycle-synchronized tobacco BY-2 (Nicotiana tabacum L. cv. Bright Yel ow 2) cel s during cel death induced by application of culture filtrates of Erwinia carotovora. The filtrate induced death in about 90%of the cel s by 24 h. Prior to cel death, vacuole shape simplified and endoplasmic actin filaments disassembled;however, the vacuoles did not rupture until after plasma membrane integrity was lost. Instead of facilitating rupture, the simplification of vacuole structure might play a role in the retrieval of membrane components needed for defense-related cel death.

  8. Cyclic Di-GMP modulates the disease progression of Erwinia amylovora.

    Science.gov (United States)

    Edmunds, Adam C; Castiblanco, Luisa F; Sundin, George W; Waters, Christopher M

    2013-05-01

    The second messenger cyclic di-GMP (c-di-GMP) is a nearly ubiquitous intracellular signal molecule known to regulate various cellular processes, including biofilm formation, motility, and virulence. The intracellular concentration of c-di-GMP is inversely governed by diguanylate cyclase (DGC) enzymes and phosphodiesterase (PDE) enzymes, which synthesize and degrade c-di-GMP, respectively. The role of c-di-GMP in the plant pathogen and causal agent of fire blight disease Erwinia amylovora has not been studied previously. Here we demonstrate that three of the five predicted DGC genes in E. amylovora (edc genes, for Erwinia diguanylate cyclase), edcA, edcC, and edcE, are active diguanylate cyclases. We show that c-di-GMP positively regulates the secretion of the main exopolysaccharide in E. amylovora, amylovoran, leading to increased biofilm formation, and negatively regulates flagellar swimming motility. Although amylovoran secretion and biofilm formation are important for the colonization of plant xylem tissues and the development of systemic infections, deletion of the two biofilm-promoting DGCs increased tissue necrosis in an immature-pear infection assay and an apple shoot infection model, suggesting that c-di-GMP negatively regulates virulence. In addition, c-di-GMP inhibited the expression of hrpA, a gene encoding the major structural component of the type III secretion pilus. Our results are the first to describe a role for c-di-GMP in E. amylovora and suggest that downregulation of motility and type III secretion by c-di-GMP during infection plays a key role in the coordination of pathogenesis.

  9. Erwinia typographi sp. nov., isolated from bark beetle (Ips typographus) gut.

    Science.gov (United States)

    Skrodenyte-Arbaciauskiene, V; Radziute, S; Stunzenas, V; Būda, V

    2012-04-01

    Gram-negative-staining bacteria that were resistant to monoterpene myrcene (7-methyl-3-methylene-1.6-octadiene, C10H16, at concentrations of up to 10 µl ml(-1) in TSB) were isolated from the gut contents of adult bark beetles Ips typographus (Coleoptera, Scolytidae). The beetles were collected from the bark of Norway spruce (Picea abies) in Lithuania. Bark beetles feed on conifers, which produce myrcene among many other defensive compounds. It has been suggested that the micro-organisms present within the beetles' guts could be involved in their resistance towards this plant defensive compound. The most resistant bacterial strains were isolated and characterized by phenotypic assays as well as fatty acid analysis, 16S rRNA gene sequencing, multilocus sequence analyses (MLSA) based on the rpoB, atpD and infB genes and DNA-DNA hybridization. Biochemical characterization indicated that the bacteria belonged to the family Enterobacteriaceae. Phylogenetic analyses of the 16S rRNA gene sequences and MLSA of the novel strains revealed that they belonged to the genus Erwinia, but represented a novel species. The dominant cellular fatty acids were C16:0 and C17:0 cyclo. The DNA G+C content was 49.1 mol%. The results obtained in this study indicated that these bacteria from the bark beetle gut represented a novel species, for which the name Erwinia typographi sp. nov. is proposed, with the type strain DSM 22678T (=Y1T=LMG 25347T).

  10. Erwinia amylovora pyrC mutant causes fire blight despite pyrimidine auxotrophy.

    Science.gov (United States)

    Ramos, L S; Sinn, J P; Lehman, B L; Pfeufer, E E; Peter, K A; McNellis, T W

    2015-06-01

    Erwinia amylovora bacteria cause fire blight disease, which affects apple and pear production worldwide. The Erw. amylovora pyrC gene encodes a predicted dihydroorotase enzyme involved in pyrimidine biosynthesis. Here, we discovered that the Erw. amylovora pyrC244::Tn5 mutant was a uracil auxotroph. Unexpectedly, the Erw. amylovora pyrC244::Tn5 mutant grew as well as the wild-type in detached immature apple and pear fruits. Fire blight symptoms caused by the pyrC244::Tn5 mutant in immature apple and pear fruits were attenuated compared to those caused by the wild-type. The pyrC244::Tn5 mutant also caused severe fire blight symptoms in apple tree shoots. A plasmid-borne copy of the wild-type pyrC gene restored prototrophy and symptom induction in apple and pear fruit to the pyrC244::Tn5 mutant. These results suggest that Erw. amylovora can obtain sufficient pyrimidine from the host to support bacterial growth and fire blight disease development, although de novo pyrimidine synthesis by Erw. amylovora is required for full symptom development in fruits. Significance and impact of the study: This study provides information about the fire blight host-pathogen interaction. Although the Erwinia amylovora pyrC mutant was strictly auxotrophic for pyrimidine, it grew as well as the wild-type in immature pear and apple fruits and caused severe fire blight disease in apple trees. This suggests that Erw. amylovora can obtain sufficient pyrimidines from host tissue to support growth and fire blight disease development. This situation contrasts with findings in some human bacterial pathogens, which require de novo pyrimidine synthesis for growth in host blood, for example.

  11. Biosynthesis of the antimetabolite 6-thioguanine in Erwinia amylovora plays a key role in fire blight pathogenesis.

    Science.gov (United States)

    Coyne, Sébastien; Chizzali, Cornelia; Khalil, Mohammed N A; Litomska, Agnieszka; Richter, Klaus; Beerhues, Ludger; Hertweck, Christian

    2013-09-27

    Sulfur for fire: The molecular basis for the biosynthesis of the antimetabolite 6-thioguanine (6TG) was unveiled in Erwinia amylovora, the causative agent of fire blight. Bioinformatics, heterologous pathway reconstitution in E. coli, and mutational analyses indicate that the protein YcfA mediates guanine thionation in analogy to 2-thiouridylase. Assays in planta and in cell cultures reveal for the first time a crucial role of 6TG in fire blight pathogenesis.

  12. Prevalence of Erwinia soft rot affecting cut foliage, Dracaena sanderiana ornamental industry and solution towards its management.

    Science.gov (United States)

    Kayalvily, Thio Desiya; Jegathambigai, V; Karunarathne, M D S D; Svinningen, Arne; Mikunthan, G

    2012-01-01

    The study was carried out under net house conditions at Green Farms Ltd, Marawila to determine the occurrence and severity of Erwinia soft rot disease in Dracaena sanderiana plants and to formulate the possible control measures. Field experiment was carried out to manage the soft rot disease in D. sanderiana plants. Three different soil treatments with vermicompost, cow dung and poultry manure were tested to manage the disease and plots without application were kept as control. Percent disease incidence, disease reduction and growth parameters were recorded and data were statistically analyzed. Higher percentage of disease reduction was observed in vermicompost (80%) treated plots than those with cow dung (60%) and poultry manure treated. Sprinkler application of water was found favorable to spread soft rot disease and watering through horse pope had lessened the disease incidence significantly. Moreover plant height, shoot and root biomass, number of leaves per plant, leaf length and leaf width were significantly high in vermicompost media. Weeding, removal of diseased leaves and plants, and avoiding sprinkler irrigation were helpful to reduce the disease spread from plant to plant. Vermicompost is the best substrate for suppression of the disease and promoting the growth of plant. Among the different water management practices tested to reduce the disease severity of Erwinia soft rot disease in D. sanderiana plants, water irrigated through the horse pipe was effective compare to sprinkler application. In-vitro experiment conducted to manage the Erwinia soft rot disease by using bio-agent, Pseudomonas fluorescens was found effective to reduce the growth of Erwinia under in-vitro conditions.

  13. Molecular characterization and pathogenicity of Erwinia spp. associated with pineapple [Ananas comosus (L. Merr.] and papaya (Carica papaya L.

    Directory of Open Access Journals (Sweden)

    Ramachandran Kogeethavani

    2015-12-01

    Full Text Available The Erwinia species are well-known pathogens of economic importance in Malaysia causing serious damage to high-value fruit crops that include pineapple [Ananas comosus (L. Merr.] and papaya (Carica papaya L..The 16S rRNA sequence using eubacteria fD1 and rP2 primers, identified two bacteria species; Dickeya zeae from pineapple heart rot, and Erwinia mallotivora from papaya dieback. Phylogenetic analysis based on the neighbor-joining method indicated that all the bacterial isolates clustered in their own taxa and formed monophyletic clades. From the pathogenicity test, all isolates of D. zeae and E. mallotivora showed pathogenic reactions on their respective host plants. Genetic variability of these isolates was assessed using repetitive sequence-based PCR (rep-PCR fingerprinting. The results indicated interspecies, and intraspecies variation in both species’ isolates. There were more polymorphic bands shown by rep-PCR fingerprints than enterobacterial repetitive intergenic consensus (ERIC and BOX- PCRs, however both species’ isolates produced distinguishable banding patterns. Unweighted pair-group method with arithmetic averages (UPGMA cluster analysis indicated that all Dickeya and Erwinia isolates from the same species were grouped in the same main cluster. Similarity among the isolates ranged from 77 to 99%. Sequencing of 16S rRNA using eubacteria fD1 and rP2 primers, and rep-PCR fingerprinting revealed diversity among Dickeya and Erwinia isolates. But this method appears to be reliable for discriminating isolates from pineapple heart rot and papaya dieback.

  14. Identification and genetics of 6-thioguanine secreted by Erwinia species and its interference with the growth of other bacteria.

    Science.gov (United States)

    Wensing, A; Gernold, M; Jock, S; Jansen, R; Geider, K

    2014-04-01

    We identified a compound in culture supernatants of Erwinia species, such as Erwinia amylovora, E. pyrifoliae, E. billingiae, E. tasmaniensis, E. persicina and E. rhapontici absorbing at 340 nm, which was associated before with the yellow pigment produced by E. amylovora on media containing copper ions. The compound was purified from E. tasmaniensis strain Et1/99 supernatants by chromatography on Dowex-1 and Dowex-50 columns and identified by HPLC/MS and NMR analysis as 6-thioguanine (6TG). Its signal at 167 Da matched with the expected molecular mass. By random mutagenesis with miniTn5, we obtained mutants defective in the genes for pyrimidine and purine metabolism. A specific gene cluster with ycf genes described by us before, absent in the corresponding region of Escherichia coli, was identified in the genome sequence of three Erwinia species and named tgs region for thioguanine synthesis. Clones of the tgs gene cluster promoted 6TG synthesis and secretion in E. coli, when the bacteria were grown in minimal medium supplemented with amino acids. 6TG was bacteriostatic for E. coli and Salmonella typhimurium strains, with cell growth resumed after prolonged incubation. Similar results were obtained with P. agglomerans strains. Bacteria from the genus Pectobacterium were barely and Rahnella or Gibbsiella species were not inhibited by 6TG. Adenine and guanine relieved the toxic effect of 6TG on E. coli. Non-producing strains were fully virulent on host plants. 6TG synthesis may help erwinias to interfere with growth of some microorganisms in the environment.

  15. Duplex real-time polymerase chain reaction reveals competition between Erwinia amylovora and E. pyrifoliae on pear blossoms.

    Science.gov (United States)

    Lehman, Susan M; Kim, Won-Sik; Castle, Alan J; Svircev, Antonet M

    2008-06-01

    Erwinia amylovora and E. pyrifoliae are the causative agents of fire blight and Asian pear blight, respectively. The pathogens are closely related, with overlapping host ranges. Data are unavailable on the current distribution of E. pyrifoliae and on the interaction between the two species when they are present together on the same host. In this study, a duplex real-time polymerase chain reaction (PCR) protocol was developed to monitor the population dynamics of E. amylovora and E. pyrifoliae on the surface of Bartlett pear blossoms. Bacterial cells washed from blossoms were used directly as the PCR template without DNA extraction. Primers and a probe based on the E. amylovora levansucrase gene detected all E. amylovora strains. All E. pyrifoliae strains, including the Japanese Erwinia strains previously described as E. amylovora, were detected with a primer and probe combination based on the E. pyrifoliae hrpW gene. Disease development and severity were not significantly different in blossoms inoculated with individual Erwinia species or with a mixture of the two species. However, E. amylovora grew to greater population sizes than did E. pyrifoliae in both single species inoculations and in mixtures, suggesting that E. amylovora has a greater competitive fitness on Bartlett pear blossoms than E. pyrifoliae.

  16. IDENTIFICATION OF THE BACTERIUM TOMATO STEM CANKER

    Directory of Open Access Journals (Sweden)

    Goner A. Shaker

    2014-01-01

    Full Text Available Diseased tomato samples were collected from green house was evaluated for isolation, pathogenicity and biochemical tests. The symptoms of the infected tomato plants were as sudden wilting after curled on leaves and necrotic streak regions developed at the crown and base of the stem and the cavities deepen and expand up and down, brown discoloration and necrosis occurring on xylem and phloem vasculer. All of ages of tomato plant were susceptible to bacteria when the weather condition favorable and immediately, seen collapse symptom on tomato plant at once fail and die. The bacterium was isolated from diseased plant in all regions on nutrient Agar; a yellow bacterium was isolated from infected tomato plant in green houses and fields in Abu-Ghraib, Rashiedia and Qanat Al-Geiaysh nurseries in Baghdad provinces of Iraq. The bacterium was found gram positive, rod-shaped, non-motile and capable an aerobic growth and based on the morphological and biochemical characteristics revealed that this bacterium belongs to: Clavibacter michiganensis subsp. michiganensis. (smith pathogenicity and hypersensitivity of the bacterium Cmm showed the disease index were 18.33, 6.66, 16.66, 5, 0% for tomato seedlings were inoculated treatments as the wounding roots, without wounding roots, crown of the stem, petiole and control respectively.

  17. 梨火疫病病原菌(Erwinia amylovora)三型分泌系统的鉴别及Erwinia spp. HrpA的分析%Identification of Type Ⅲ Secration System in Erwinia amylovora and Analyse of HrpA in Erwinia spp.

    Institute of Scientific and Technical Information of China (English)

    朱勃; 金谷雷; 怀雁; 谢关林

    2009-01-01

    [目的]明确梨火疫病病原菌(Erwinia amylovora)三型分泌系统(type Ⅲ secretion system,TTSS)的所在区域、相关基因及Erwiaia spp.的HrpA(hypersensitive response and pathogenicity gene A)选择压水平.[方法]利用生物信息学方法,通过BLAST程序对梨火疫病菌基因组的核苷酸数据库进行同源比对,同时对Erwiaia spp.与宿主互作的表面蛋白HrpA进行选择压分析.[结果] TTSS分析结果显示在40 kb大小的毒力岛上有27个TTSS相关基因.通过与看家基因及毒力岛上的致病性相关的重要基因HrpN的比较、选择压分析、多序列比较,发现HrpA基因处于较强的选择压作用下,且HrpA蛋白N端主要受正向选择,C端主要受净化选择.NJ法构建的HrpA系统发育树显示Erwinia spp.形成两个明显的分支,表明HrpA基因可能在种间分化后产生了不同的选择压变化.[结论]梨火疫病病原菌基因组3.14-3.18 Mb为其TTSS分布区域.该病原菌通过TTSS侵染寄主植物,在其病原菌表面有一种类鞭毛结构的TTSS蛋白HrpA,HrpA作为传输者起着将效应分子输送到宿主内部的功能,在进化上受到了较强的选择压影响.

  18. Zymomonas mobilis: a bacterium for ethanol production

    Energy Technology Data Exchange (ETDEWEB)

    Baratti, J.C.; Bu' Lock, J.D.

    1986-01-01

    Zymomonas mobilis is a facultative anaerobic gram negative bacterium first isolated in tropical countries from alcoholic beverages like the African palm wine, the Mexican pulque and also as a contaminant of cider (cider sickness) or beer in the European countries. It is one of the few facultative anaerobic bacteria degrading glucose by the Entner-Doudoroff pathway usually found in strictly aerobic microorganisms. Some work was devoted to this bacterium in the 50s and 60s and was reviewed by Swings and De Ley in their classical paper published in 1977. During the 70s there was very little work on the bacterium until 1979 and the first report by the Australian group of P.L. Rogers on the great potentialities of Z. mobilis for ethanol production. At that time the petroleum crisis had led the developed countries to search for alternative fuel from renewable resources. The Australian group clearly demonstrated the advantages of the bacterium compared to the yeasts traditionally used for the alcoholic fermentation. As a result, there was a considerable burst in the Zymomonas literature which started from nearly zero in the late 70s to attain 70 papers published in the field in 1984. In this article, papers published from 1982 to 1986 are reviewed.

  19. Characterization of a new ViI-like Erwinia amylovora bacteriophage phiEa2809.

    Science.gov (United States)

    Lagonenko, Alexander L; Sadovskaya, Olga; Valentovich, Leonid N; Evtushenkov, Anatoly N

    2015-04-01

    Erwinia amylovora is a Gram-negative plant pathogenic bacteria causing fire blight disease in many Rosaceae species. A novel E. amylovora bacteriophage, phiEa2809, was isolated from symptomless apple leaf sample collected in Belarus. This phage was also able to infect Pantoea agglomerans strains. The genome of phiEa2809 is a double-stranded linear DNA 162,160 bp in length, including 145 ORFs and one tRNA gene. The phiEa2809 genomic sequence is similar to the genomes of the Serratia plymutica phage MAM1, Shigella phage AG-3, Dickeya phage vB DsoM LIMEstone1 and Salmonella phage ViI and lacks similarity to described E. amylovora phage genomes. Based on virion morphology (an icosahedral head, long contractile tail) and genome structure, phiEa2809 was classified as a member of Myoviridae, ViI-like bacteriophages group. PhiEa2809 is the firstly characterized ViI-like bacteriophage able to lyse E. amylovora.

  20. Cell surface attachment structures contribute to biofilm formation and xylem colonization by Erwinia amylovora.

    Science.gov (United States)

    Koczan, Jessica M; Lenneman, Bryan R; McGrath, Molly J; Sundin, George W

    2011-10-01

    Biofilm formation plays a critical role in the pathogenesis of Erwinia amylovora and the systemic invasion of plant hosts. The functional role of the exopolysaccharides amylovoran and levan in pathogenesis and biofilm formation has been evaluated. However, the role of biofilm formation, independent of exopolysaccharide production, in pathogenesis and movement within plants has not been studied previously. Evaluation of the role of attachment in E. amylovora biofilm formation and virulence was examined through the analysis of deletion mutants lacking genes encoding structures postulated to function in attachment to surfaces or in cellular aggregation. The genes and gene clusters studied were selected based on in silico analyses. Microscopic analyses and quantitative assays demonstrated that attachment structures such as fimbriae and pili are involved in the attachment of E. amylovora to surfaces and are necessary for the production of mature biofilms. A time course assay indicated that type I fimbriae function earlier in attachment, while type IV pilus structures appear to function later in attachment. Our results indicate that multiple attachment structures are needed for mature biofilm formation and full virulence and that biofilm formation facilitates entry and is necessary for the buildup of large populations of E. amylovora cells in xylem tissue.

  1. EDS1 contributes to nonhost resistance of Arabidopsis thaliana against Erwinia amylovora.

    Science.gov (United States)

    Moreau, Manon; Degrave, Alexandre; Vedel, Régine; Bitton, Frédérique; Patrit, Oriane; Renou, Jean-Pierre; Barny, Marie-Anne; Fagard, Mathilde

    2012-03-01

    Erwinia amylovora causes fire blight in rosaceous plants. In nonhost Arabidopsis thaliana, E. amylovora triggers necrotic symptoms associated with transient bacterial multiplication, suggesting either that A. thaliana lacks a susceptibility factor or that it actively restricts E. amylovora growth. Inhibiting plant protein synthesis at the time of infection led to an increase in necrosis and bacterial multiplication and reduced callose deposition, indicating that A. thaliana requires active protein synthesis to restrict E. amylovora growth. Analysis of the callose synthase-deficient pmr4-1 mutant indicated that lack of callose deposition alone did not lead to increased sensitivity to E. amylovora. Transcriptome analysis revealed that approximately 20% of the genes induced following E. amylovora infection are related to defense and signaling. Analysis of mutants affected in NDR1 and EDS1, two main components of the defense-gene activation observed, revealed that E. amylovora multiplied ten times more in the eds1-2 mutant than in the wild type but not in the ndr1-1 mutant. Analysis of mutants affected in three WRKY transcription factors showing EDS1-dependent activation identified WRKY46 and WRKY54 as positive regulators and WRKY70 as a negative regulator of defense against E. amylovora. Altogether, we show that EDS1 is a positive regulator of nonhost resistance against E. amylovora in A. thaliana and hypothesize that it controls the production of several effective defenses against E. amylovora through the action of WRKY46 and WRKY54, while WRKY70 acts as a negative regulator.

  2. Biomolecular characterization of the levansucrase of Erwinia amylovora, a promising biocatalyst for the synthesis of fructooligosaccharides.

    Science.gov (United States)

    Caputi, Lorenzo; Nepogodiev, Sergey A; Malnoy, Mickael; Rejzek, Martin; Field, Robert A; Benini, Stefano

    2013-12-18

    Erwinia amylovora is a plant pathogen that affects Rosaceae, such as apple and pear. In E. amylovora the fructans, produced by the action of a levansucrase (EaLsc), play a role in virulence and biofilm formation. Fructans are bioactive compounds, displaying health-promoting properties in their own right. Their use as food and feed supplements is increasing. In this study, we investigated the biomolecular properties of EaLsc using HPAEC-PAD, MALDI-TOF MS, and spectrophotometric assays. The enzyme, which was heterologously expressed in Escherichia coli in high yield, was shown to produce mainly fructooligosaccharides (FOSs) with a degree of polymerization between 3 and 6. The kinetic properties of EaLsc were similar to those of other phylogenetically related Gram-negative bacteria, but the good yield of FOSs, the product spectrum, and the straightforward production of the enzyme suggest that EaLsc is an interesting biocatalyst for future studies aimed at producing tailor-made fructans.

  3. Global small RNA chaperone Hfq and regulatory small RNAs are important virulence regulators in Erwinia amylovora.

    Science.gov (United States)

    Zeng, Quan; McNally, R Ryan; Sundin, George W

    2013-04-01

    Hfq is a global small RNA (sRNA) chaperone that interacts with Hfq-regulated sRNAs and functions in the posttranscriptional regulation of gene expression. In this work, we identified Hfq to be a virulence regulator in the Gram-negative fire blight pathogen Erwinia amylovora. Deletion of hfq in E. amylovora Ea1189 significantly reduced bacterial virulence in both immature pear fruits and apple shoots. Analysis of virulence determinants in strain Ea1189Δhfq showed that Hfq exerts pleiotropic regulation of amylovoran exopolysaccharide production, biofilm formation, motility, and the type III secretion system (T3SS). Further characterization of biofilm regulation by Hfq demonstrated that Hfq limits bacterial attachment to solid surfaces while promoting biofilm maturation. Characterization of T3SS regulation by Hfq revealed that Hfq positively regulates the translocation and secretion of the major type III effector DspE and negatively controls the secretion of the putative translocator HrpK and the type III effector Eop1. Lastly, 10 Hfq-regulated sRNAs were identified using a computational method, and two of these sRNAs, RprA and RyhA, were found to be required for the full virulence of E. amylovora.

  4. Differential lysine acetylation profiles of Erwinia amylovora strains revealed by proteomics.

    Science.gov (United States)

    Wu, Xia; Vellaichamy, Adaikkalam; Wang, Dongping; Zamdborg, Leonid; Kelleher, Neil L; Huber, Steven C; Zhao, Youfu

    2013-02-21

    Protein lysine acetylation (LysAc) has recently been demonstrated to be widespread in E. coli and Salmonella, and to broadly regulate bacterial physiology and metabolism. However, LysAc in plant pathogenic bacteria is largely unknown. Here we first report the lysine acetylome of Erwinia amylovora, an enterobacterium causing serious fire blight disease of apples and pears. Immunoblots using generic anti-lysine acetylation antibodies demonstrated that growth conditions strongly affected the LysAc profiles in E. amylovora. Differential LysAc profiles were also observed for two E. amylovora strains, known to have differential virulence in plants, indicating translational modification of proteins may be important in determining virulence of bacterial strains. Proteomic analysis of LysAc in two E. amylovora strains identified 141 LysAc sites in 96 proteins that function in a wide range of biological pathways. Consistent with previous reports, 44% of the proteins are involved in metabolic processes, including central metabolism, lipopolysaccharide, nucleotide and amino acid metabolism. Interestingly, for the first time, several proteins involved in E. amylovora virulence, including exopolysaccharide amylovoran biosynthesis- and type III secretion-associated proteins, were found to be lysine acetylated, suggesting that LysAc may play a major role in bacterial virulence. Comparative analysis of LysAc sites in E. amylovora and E. coli further revealed the sequence and structural commonality for LysAc in the two organisms. Collectively, these results reinforce the notion that LysAc of proteins is widespread in bacterial metabolism and virulence.

  5. Lipopolysaccharide biosynthesis genes discriminate between Rubus- and Spiraeoideae-infective genotypes of Erwinia amylovora.

    Science.gov (United States)

    Rezzonico, Fabio; Braun-Kiewnick, Andrea; Mann, Rachel A; Rodoni, Brendan; Goesmann, Alexander; Duffy, Brion; Smits, Theo H M

    2012-10-01

    Comparative genomic analysis revealed differences in the lipopolysaccharide (LPS) biosynthesis gene cluster between the Rubus-infecting strain ATCC BAA-2158 and the Spiraeoideae-infecting strain CFBP 1430 of Erwinia amylovora. These differences corroborate rpoB-based phylogenetic clustering of E. amylovora into four different groups and enable the discrimination of Spiraeoideae- and Rubus-infecting strains. The structure of the differences between the two groups supports the hypothesis that adaptation to Rubus spp. took place after species separation of E. amylovora and E. pyrifoliae that contrasts with a recently proposed scenario, based on CRISPR data, in which the shift to domesticated apple would have caused an evolutionary bottleneck in the Spiraeoideae-infecting strains of E. amylovora which would be a much earlier event. In the core region of the LPS biosynthetic gene cluster, Spiraeoideae-infecting strains encode three glycosyltransferases and an LPS ligase (Spiraeoideae-type waaL), whereas Rubus-infecting strains encode two glycosyltransferases and a different LPS ligase (Rubus-type waaL). These coding domains share little to no homology at the amino acid level between Rubus- and Spiraeoideae-infecting strains, and this genotypic difference was confirmed by polymerase chain reaction analysis of the associated DNA region in 31 Rubus- and Spiraeoideae-infecting strains. The LPS biosynthesis gene cluster may thus be used as a molecular marker to distinguish between Rubus- and Spiraeoideae-infecting strains of E. amylovora using primers designed in this study.

  6. Characterization of Erwinia amylovora strains from Bulgaria by pulsed-field gel electrophoresis.

    Science.gov (United States)

    Atanasova, Iliana; Urshev, Zoltan; Hristova, Petya; Bogatzevska, Nevena; Moncheva, Penka

    2012-01-01

    The aim of this study was to characterize genetically Bulgarian Erwinia amylovora strains using pulsed-field gel electrophoresis (PFGE) analysis. Fifty E. amylovora strains isolated from different hosts, locations, as well as in different years were analysed by PFGE after XbaI, SpeI, and XhoI digestion of the genomic DNA. The strains were distributed into four groups according to their XbaI-generated profile. About 82% of the strains displayed a PFGE profile identical to that of type Pt2. Three strains belonged to the Central Europe Pt1 type. Two new PFGE profiles, not reported so far, were established--one for a strain isolated from Malus domestica and another for all Fragaria spp. strains. The same grouping of the strains was obtained after analysis of the SpeI digestion patterns. On the basis of PFGE profiles, after XbaI and SpeI digestion, a genetic differentiation between the strains associated with subfamily Maloideae and subfamily Rosoideae was revealed. The presence of more than one PFGE profile in the population of E. amylovora in Bulgaria suggests a multiple source of inoculum.

  7. Involvement of N-acylhomoserine lactones throughout plant infection by Erwinia carotovora subsp. atroseptica (Pectobacterium atrosepticum).

    Science.gov (United States)

    Smadja, Bruno; Latour, Xavier; Faure, Denis; Chevalier, Sylvie; Dessaux, Yves; Orange, Nicole

    2004-11-01

    Erwinia carotovora subsp. atroseptica is responsible for potato blackleg disease in the field and tuber soft rot during crop storage. The process leading to the disease occurs in two phases: a primary invasion step followed by a maceration step. Bacteria-to-bacteria communication is associated with a quorum-sensing (QS) process based on the production of N-acylhomoserine lactones (HSL). The role of HSL throughout plant infection was analyzed. To this purpose, HSL produced by a specific E. carotovora subsp. atroseptica wild-type strain, which was particularly virulent on potato, were identified. A derivative of this strain that expressed an HSL lactonase gene and produced low amounts of HSL was generated. The comparison of these strains allowed the evaluation of the role of HSL and QS in disease establishment and development. Bacterial growth and motility; activity of proteins secreted by type I, II, and III systems; and hypersensitive and maceration reactions were evaluated. Results indicated that HSL production and QS regulate only those traits involved in the second stage of the host plant infection (i.e., tissue maceration) and hypersensitive response in nonhost tobacco plants. Therefore, the use of QS quenching strategies for biological control in E. carotovora subsp. atroseptica cannot prevent initial infection and multiplication of this pathogen.

  8. Antimicrobial and anti-pathogenic activity of some thioureides derivatives against Erwinia amylovora phytopathogenic strains.

    Science.gov (United States)

    Măruţescu, Luminiţa; Niţulescu, Mihai-George; Bucur, Marcela; Diţu, Lia-Mara; Mihăescu, Grigore; Lazăr, Veronica; Sesan, Tatiana

    2011-01-01

    A series of N-(1-methyl-1 Hpyrazole-4-carbonyl)-thiourea derivatives were assessed for their in vitro antimicrobial and anti-pathogenic activity against twenty-two strains of Erwinia amylovora isolated from different regions in Romania. The compounds were solubilised in dimethylsulfoxide and screened for their in vitro antimicrobial activity. The qualitative screening of the susceptibility spectra of various strains to the compounds was performed by adapted diffusion techniques (distribution of the tested compound solution directly on the solid medium previously seeded with the bacterial inoculums). The quantitative assay of the minimal inhibitory concentration (MIC, microg/mL) was based on liquid medium two-fold microdilutions. The subinhibitory concentrations of the tested substances were investigated for their influence on biofilm development on inert substrata. The present study showed that six new thiourea compounds exhibited a low antibacterial activity (MIC values > 500 microg/ml), but the subinhibitory concentrations inhibited the biofilm development on inert substrata. Thus, these results could suggest the usefulness of the tested compounds as control agents for preventing the first stage (colonization) of the infection with the fire blight pathogen.

  9. Cloning and characterization of a sucrose isomerase from Erwinia rhapontici NX-5 for isomaltulose hyperproduction.

    Science.gov (United States)

    Li, Sha; Cai, Heng; Qing, Yujia; Ren, Ben; Xu, Hong; Zhu, Hongyang; Yao, Jun

    2011-01-01

    The sucrose isomerase (SIase) gene from an efficient strain of Erwinia rhapontici NX-5 for isomaltulose hyperproduction was cloned and overexpressed in Escherichia coli. Protein sequence alignment revealed that SIase was a member of the glycoside hydrolase 13 family. The molecular mass of the purified recombinant protein was estimated at 66 kDa by SDS-PAGE. The SIase had an optimal pH and temperature of 5.0 and 30 °C, respectively, with a K (m) of 257 mmol/l and V (max) of 48.09 μmol/l/s for sucrose. To the best of our knowledge, the recombinant SIase has the most acidic optimum pH for isomaltulose synthesis. When the recombinant E. coli (pET22b- palI) cells were used for isomaltulose synthesis, almost complete conversion of sucrose (550 g/l solution) to isomaltulose was achieved in 1.5 h with high isomaltulose yields (87%). The immobilized E. coli cells remained stable for more than 30 days in a "batch"-type enzyme reactor. This indicated that the recombinant SIase could continuously and efficiently produce isomaltulose.

  10. Purification and characterization of a highly selective sucrose isomerase from Erwinia rhapontici NX-5.

    Science.gov (United States)

    Ren, Ben; Li, Sha; Xu, Hong; Feng, Xiao-Hai; Cai, Heng; Ye, Qi

    2011-06-01

    A highly selective sucrose isomerase (SIase) was purified to homogeneity from the cell-free extract of Erwinia rhapontici NX-5 with a recovery of 27.7% and a fold purification of 213.6. The purified SIase showed a high specific activity of 427.1 U mg(-1) with molecular weight of 65.6 kDa. The K (m) for sucrose was 222 mM while V (max) was 546 U mg(-1). The optimum pH and temperature for SIase activity were 6.0 and 30 °C, respectively. The purified SIase was stable in the temperature range of 10-40 °C and retained 65% of the enzyme activity after 2 weeks' storage at 30 °C. The SIase activity was enhanced by Mg(2+) and Mn(2+), inhibited by Ca(2+), Cu(2+), Zn(2+), and Co(2+), completely inhibited by Hg(2+) and Ag(2+). The purified SIase was strongly inhibited by SDS, while partially inhibited by dimethylformamide, tetrahydrofuran, and PMSF. Additionally, glucose and fructose acted as competitive inhibitors for purified SIase.

  11. Modified inoculation and disease assessment methods reveal host specificity in Erwinia tracheiphila-Cucurbitaceae interactions.

    Science.gov (United States)

    Nazareno, Eric S; Dumenyo, C Korsi

    2015-12-01

    We conducted a greenhouse trial to determine specific compatible interactions between Erwinia tracheiphila strains and cucurbit host species. Using a modified inoculation system, E. tracheiphila strains HCa1-5N, UnisCu1-1N, and MISpSq-N were inoculated to cucumber (Cucumis sativus) cv. 'Sweet Burpless', melon (Cucumis melo) cv. 'Athena Hybrid', and squash (Cucubita pepo) cv. 'Early Summer Crookneck'. We observed symptoms and disease progression for 30 days; recorded the number of days to wilting of the inoculated leaf (DWIL), days to wilting of the whole plant (DWWP), and days to death of the plant (DDP). We found significant interactions between host cultivar and pathogen strains, which imply host specificity. Pathogen strains HCa1-5N and UnisCu1-1N isolated from Cucumis species exhibited more virulence in cucumber and melon than in squash, while the reverse was true for strain MISpSq-N, an isolate from Cucurbita spp. Our observations confirm a previous finding that E. tracheiphila strains isolated from Cucumis species were more virulent on Cucumis hosts and those from Cucubita were more virulent on Cucubita hosts. This confirmation helps in better understanding the pathosystem and provides baseline information for the subsequent development of new disease management strategies for bacterial wilt. We also demonstrated the efficiency of our modified inoculation and disease scoring methods.

  12. Sucrose isomerase and its mutants from Erwinia rhapontici can synthesise α-arbutin.

    Science.gov (United States)

    Zhou, Xing; Zheng, Yuantao; Wei, Xingming; Yang, Kedi; Yang, Xiangkai; Wang, Yuting; Xu, Liming; Du, Liqin; Huang, Ribo

    2011-10-01

    Sucrose isomerase (SI) from Erwinia rhapontici is an intramolecular isomerase that is normally used to synthesise isomaltulose from sucrose by a mechanism of intramolecular transglycosylation. In this study, it was found that SI could synthesise α-arbutin using hydroquinone and sucrose as substrates, via an intermolecular transglycosylation reaction. Five phenylalanine residues (F185, F186, F205, F297, and F321) in the catalytic pocket of SI were chosen for sitedirected mutagenesis. Mutants F185I, F321I, and F321W, whose hydrolytic activities were enhanced after the mutation, could synthesise α-arbutin through intermolecular transglycosylation with a more than two-fold increase in the molar transfer ratio compared with wild type SI. The F297A mutant showed a strong ability to synthesise a novel α-arbutin derivative and a four-fold increase in its specific activity for intermolecular transglycosylation over the wild type. Our findings may lead to a new way to synthesise novel glucoside products such as α-arbutin derivatives by simply manipulating the Phe residues in the catalytic pocket. From the structure superposition, our strategy of manipulating these Phe residues may be applicable to other similar transglycosylating enzymes.

  13. Morphological and biochemical characterization of Erwinia amylovora-induced hypersensitive cell death in apple leaves.

    Science.gov (United States)

    Iakimova, Elena T; Sobiczewski, Piotr; Michalczuk, Lech; Węgrzynowicz-Lesiak, Elżbieta; Mikiciński, Artur; Woltering, Ernst J

    2013-02-01

    In attached apple leaves, spot-inoculated with Erwinia amylovora, the phenotypic appearance of the hypersensitive response (HR) and the participation of ethylene, reactive oxygen species (ROS) and of vacuolar processing enzyme (VPE) (a plant caspase-1-like protease) were analysed. The HR in both the resistant and susceptible genotypes expressed a similar pattern of distinguishable micro HR lesions that progressed into confined macro HR lesions. The HR symptoms in apple were compared to those in non-host tobacco. The morphology of dead cells (protoplast shrinkage and retraction from cell wall) in apple leaves resembled necrotic programmed cell death (PCD). Lesion formation in both cv. Free Redstar (resistant) and cv. Idared (highly susceptible) was preceded by ROS accumulation and elevation of ethylene levels. Treatment of infected leaves with an inhibitor of ethylene synthesis led to a decrease of ethylene emission and suppression of lesion development in both cultivars. In the resistant but not in the susceptible apple cultivar an early and late increase in VPE gene expression was detected. This suggests that VPE might be an underlying component of the response to E. amylovora in resistant apple cultivars. The findings show that in the studied pathosystem the cell death during the HR proceeds through a signal transduction cascade in which ROS, ethylene and VPE pathways play a role.

  14. Insecticidal Bacillus thuringiensis silences Erwinia carotovora virulence by a new form of microbial antagonism, signal interference.

    Science.gov (United States)

    Dong, Yi-Hu; Zhang, Xi-Fen; Xu, Jin-Ling; Zhang, Lian-Hui

    2004-02-01

    It is commonly known that bacteria may produce antibiotics to interfere with the normal biological functions of their competitors in order to gain competitive advantages. Here we report that Bacillus thuringiensis suppressed the quorum-sensing-dependent virulence of plant pathogen Erwinia carotovora through a new form of microbial antagonism, signal interference. E. carotovora produces and responds to acyl-homoserine lactone (AHL) quorum-sensing signals to regulate antibiotic production and expression of virulence genes, whereas B. thuringiensis strains possess AHL-lactonase, which is a potent AHL-degrading enzyme. B. thuringiensis did not seem to interfere with the normal growth of E. carotovora; rather, it abolished the accumulation of AHL signal when they were cocultured. In planta, B. thuringiensis significantly decreased the incidence of E. carotovora infection and symptom development of potato soft rot caused by the pathogen. The biocontrol efficiency is correlated with the ability of bacterial strains to produce AHL-lactonase. While all the seven AHL-lactonase-producing B. thuringiensis strains provided significant protection against E. carotovora infection, Bacillus fusiformis and Escherichia coli strains that do not process AHL-degradation enzyme showed little effect in biocontrol. Mutation of aiiA, the gene encoding AHL-lactonase in B. thuringiensis, resulted in a substantial decrease in biocontrol efficacy. These results suggest that signal interference mechanisms existing in natural ecosystems could be explored as a new version of antagonism for prevention of bacterial infections.

  15. First report of Pantoea ananatis (Syn. Erwinia uredovora) being associated with peanut rust in Georgia

    Science.gov (United States)

    Peanut rust is caused by the fungus Puccinia arachidis. This disease, if not treated can cause severe damage and defoliation. While sequencing DNA of urediniospores of the rust fungus, BLAST analysis detected many sequences corresponding to the bacterial species Pantoea ananatis. This bacterium, ...

  16. Novel Waddlia Intracellular Bacterium in Artibeus intermedius Fruit Bats, Mexico.

    Science.gov (United States)

    Pierlé, Sebastián Aguilar; Morales, Cirani Obregón; Martínez, Leonardo Perea; Ceballos, Nidia Aréchiga; Rivero, Juan José Pérez; Díaz, Osvaldo López; Brayton, Kelly A; Setién, Alvaro Aguilar

    2015-12-01

    An intracellular bacterium was isolated from fruit bats (Artibeus intermedius) in Cocoyoc, Mexico. The bacterium caused severe lesions in the lungs and spleens of bats and intracytoplasmic vacuoles in cell cultures. Sequence analyses showed it is related to Waddlia spp. (order Chlamydiales). We propose to call this bacterium Waddlia cocoyoc.

  17. Production of glucosyltransferase by Erwinia sp. using experimental design and response surface methodology Produção de glicosiltransferase por Erwinia sp. utilizando planejamento experimental e metodologia de superfície de resposta

    Directory of Open Access Journals (Sweden)

    Haroldo Yukio Kawaguti

    2005-09-01

    Full Text Available Glucosyltransferase produced by strain Erwinia sp. is an intracellular enzyme that catalyzes the formation of isomaltulose from sucrose. Isomaltulose is a non-cariogenic reducing dissacharide commercially used in foods. Response surface methodology and 2³-factorial central composite design were employed to optimize a fermentation medium for the production of glucosyltransferase by Erwinia sp. in shaken flasks at 200 rpm and 30ºC. The three variables involved in this study were sugar cane molasses (SCM, corn steep liquor (CSL and yeast extract Prodex Lac SD (YEP. The statistical analysis of the results showed that, in the range studied, all the factors had a significant effect on glucosyltransferase production and the optimum medium composition for enzyme production was (in g l-1 SCM-100, CSL-60 and YEP-8, which lead to a glucosyltransferase activity of 6.65 U mL-1.A glicosiltransferase obtida pela linhagem Erwinia sp. é uma enzima intracelular que catalisa a conversão de sacarose em isomaltulose. A isomaltulose é um dissacarídeo redutor, não cariogênico e comercialmente utilizado em alimentos como substituto da sacarose. A metodologia de superfície de resposta e planejamento fatorial composto central-2³ foram utilizados para otimizar o meio de cultivo para a produção de glicosiltransferase de Erwinia sp. em frascos sob agitação a 200 rpm e 30ºC. As três variáveis independentes envolvidas no estudo foram o melaço de cana de açúcar, a água de maceração de milho e o extrato de levedura Prodex Lac SD. As análises estatísticas dos resultados mostraram que, dentro da faixa estudada das concentrações dos componentes de meio de cultivo, todas as variáveis apresentaram efeito significativo na produção de glicosiltransferase. O meio de cultivo otimizado foi composto de 100 gL-1 de melaço de cana de açúcar, 60 gL-1 de água de maceração de milho e 8 gL-1 de extrato de levedura Prodex Lac SD, apresentando atividade de

  18. Fire blight (Erwinia amylovora) of rosaceous plants. Pathogen virulence and selection and characterization of biological control agents

    OpenAIRE

    Cabrefiga Olamendi, Jordi

    2004-01-01

    El fuego bacteriano, causado por Erwinia amylovora, es una enfermedad muy importante a nivel comercial y económico porque afecta a plantas de la familia de las rosáceas y es especialmente agresiva en manzano (Pyrus malus) y peral (Pyrus communis), así como en plantas ornamentales (Crataegus, Cotoneaster o Pyracantha). Esta enfermedad está distribuida por todo el mundo en zonas climáticas templadas de Amércia del Norte, Nueva Zelanda, Japón, Israel, Turquí y Europa. En España, el fuego bacter...

  19. Molecular cloning and characterization of an Erwinia carotovora subsp. carotovora pectin lyase gene that responds to DNA-damaging agents.

    OpenAIRE

    McEvoy, J L; Murata, H.; Chatterjee, A. K.

    1990-01-01

    recA-mediated production of pectin lyase (PNL) and the bacteriocin carotovoricin occurs in Erwinia carotovora subsp. carotovora 71 when this organism is subjected to agents that damage or inhibit the synthesis of DNA. The structural gene pnlA was isolated from a strain 71 cosmid gene library following mobilization of the cosmids into a moderate PNL producer, strain 193. The cosmid complemented pnl::Tn5 but not ctv::Tn5 mutations. A constitutive level of PNL activity was detected in RecA+ and ...

  20. Antagonistic potential of Pseudomonas graminis 49M against Erwinia amylovora, the causal agent of fire blight.

    Science.gov (United States)

    Mikiciński, Artur; Sobiczewski, Piotr; Puławska, Joanna; Malusa, Eligio

    2016-08-01

    In a previous study (Mikiciński et al. in Eur J Plant Pathol, doi: 10.1007/s10658-015-0837-y , 2015), we described the characterization of novel strain 49M of Pseudomonas graminis, isolated from the phyllosphere of apple trees in Poland showing a good protective activity against fire blight on different organs of host plants. We now report investigations to clarify the basis for this activity. Strain 49M was found to produce siderophores on a medium containing complex CAS-Fe(3+) and HDTMA, but was not able to produce N-acyl homoserine lactones (AHLs). Moreover, it formed a biofilm on polystyrene and polyvinyl chloride (PVC) surfaces. Strain 49M gave a positive reaction in PCR with primers complementary to gacA, the regulatory gene influencing the production of several secondary metabolites including antibiotics. The genes prnD (encoding pyrrolnitrin), pltC, pltB (pyoluteorin), phlD (2,4-diacetyl-phloroglucinol) and phzC as well as phzD (and their homologs phzF and phzA encoding phenazine), described for antagonistic fluorescent pseudomonads, however, were not detected. Research into the biotic relationship between strain 49M and Erwinia amylovora strain Ea659 on five microbiological media showed that this strain clearly inhibited the growth of the pathogen on King's B and nutrient agar with glycerol media, to a very small extent on nutrient agar with sucrose, and not at all on Luria-Bertani agar. On medium 925, strain 49M even stimulated E. amylovora growth. The addition of ferric chloride to King's B resulted in the loss of its inhibitory ability. Testing the survival of 49M in vitro showed its resistance to drought, greater than that of E. amylovora.

  1. AmyR is a novel negative regulator of amylovoran production in Erwinia amylovora.

    Directory of Open Access Journals (Sweden)

    Dongping Wang

    Full Text Available In this study, we attempted to understand the role of an orphan gene amyR in Erwinia amylovora, a functionally conserved ortholog of ybjN in Escherichia coli, which has recently been characterized. Amylovoran, a high molecular weight acidic heteropolymer exopolysaccharide, is a virulent factor of E. amylovora. As reported earlier, amylovoran production in an amyR knockout mutant was about eight-fold higher than that in the wild type (WT strain of E. amylovora. When a multicopy plasmid containing the amyR gene was introduced into the amyR mutant or WT strains, amylovoran production was strongly inhibited. Furthermore, amylovoran production was also suppressed in various amylovoran-over-producing mutants, such as grrSA containing multicopies of the amyR gene. Consistent with amylovoran production, an inverse correlation was observed between in vitro expression of amyR and that of amylovoran biosynthetic genes. However, both the amyR knockout mutant and over-expression strains showed reduced levan production, another exopolysaccharide produced by E. amylovora. Virulence assays demonstrated that while the amyR mutant was capable of inducing slightly greater disease severity than that of the WT strain, strains over-expressing the amyR gene did not incite disease on apple shoots or leaves, and only caused reduced disease on immature pear fruits. Microarray studies revealed that amylovoran biosynthesis and related membrane protein-encoding genes were highly expressed in the amyR mutant, but down-regulated in the amyR over-expression strains in vitro. Down-regulation of amylovoran biosynthesis genes in the amyR over-expression strain partially explained why over-expression of amyR led to non-pathogenic or reduced virulence in vivo. These results suggest that AmyR plays an important role in regulating exopolysaccharide production, and thus virulence in E. amylovora.

  2. Genome-wide identification of genes regulated by the Rcs phosphorelay system in Erwinia amylovora.

    Science.gov (United States)

    Wang, Dongping; Qi, Mingsheng; Calla, Bernarda; Korban, Schuyler S; Clough, Steven J; Cock, Peter J A; Sundin, George W; Toth, Ian; Zhao, Youfu

    2012-01-01

    The exopolysaccharide amylovoran is one of the major pathogenicity factors in Erwinia amylovora, the causal agent of fire blight of apples and pears. We have previously demonstrated that the RcsBCD phosphorelay system is essential for virulence by controlling amylovoran biosynthesis. We have also found that the hybrid sensor kinase RcsC differentially regulates amylovoran production in vitro and in vivo. To further understand how the Rcs system regulates E. amylovora virulence gene expression, we conducted genome-wide microarray analyses to determine the regulons of RcsB and RcsC in liquid medium and on immature pear fruit. Array analyses identified a total of 648 genes differentially regulated by RcsCB in vitro and in vivo. Consistent with our previous findings, RcsB acts as a positive regulator in both conditions, while RcsC positively controls expression of amylovoran biosynthetic genes in vivo but negatively controls expression in vitro. Besides amylovoran biosynthesis and regulatory genes, cell-wall and cell-envelope (membrane) as well as regulatory genes were identified as the major components of the RcsBC regulon, including many novel genes. We have also demonstrated that transcripts of rcsA, rcsC, and rcsD genes but not the rcsB gene were up-regulated when bacterial cells were grown in minimal medium or following infection of pear fruits compared with those grown in Luria Bertani medium. Furthermore, using the genome of E. amylovora ATCC 49946, a hidden Markov model predicted 60 genes with a candidate RcsB binding site in the intergenic region, 28 of which were identified in the microarray assay. Based on these findings as well as previous reported data, a working model has been proposed to illustrate how the Rcs phosphorelay system regulates virulence gene expression in E. amylovora.

  3. The fire blight pathogen Erwinia amylovora requires the rpoN gene for pathogenicity in apple.

    Science.gov (United States)

    Ramos, Laura S; Lehman, Brian L; Sinn, Judith P; Pfeufer, Emily E; Halbrendt, Noemi O; McNellis, Timothy W

    2013-10-01

    RpoN is a σ(54) factor regulating essential virulence gene expression in several plant pathogenic bacteria, including Pseudomonas syringae and Pectobacterium carotovorum. In this study, we found that mutation of rpoN in the fire blight pathogen Erwinia amylovora caused a nonpathogenic phenotype. The E. amylovora rpoN Tn5 transposon mutant rpoN1250::Tn5 did not cause fire blight disease symptoms on shoots of mature apple trees. In detached immature apple fruits, the rpoN1250::Tn5 mutant failed to cause fire blight disease symptoms and grew to population levels 12 orders of magnitude lower than the wild-type. In addition, the rpoN1250::Tn5 mutant failed to elicit a hypersensitive response when infiltrated into nonhost tobacco plant leaves, and rpoN1250::Tn5 cells failed to express HrpN protein when grown in hrp (hypersensitive response and pathogenicity)-inducing liquid medium. A plasmid-borne copy of the wild-type rpoN gene complemented all the rpoN1250::Tn5 mutant phenotypes tested. The rpoN1250::Tn5 mutant was prototrophic on minimal solid and liquid media, indicating that the rpoN1250::Tn5 nonpathogenic phenotype was not caused by a defect in basic metabolism or growth. This study provides clear genetic evidence that rpoN is an essential virulence gene of E. amylovora, suggesting that rpoN has the same function in E. amylovora as in P. syringae and Pe. carotovorum.

  4. AmyR is a novel negative regulator of amylovoran production in Erwinia amylovora.

    Science.gov (United States)

    Wang, Dongping; Korban, Schuyler S; Pusey, P Lawrence; Zhao, Youfu

    2012-01-01

    In this study, we attempted to understand the role of an orphan gene amyR in Erwinia amylovora, a functionally conserved ortholog of ybjN in Escherichia coli, which has recently been characterized. Amylovoran, a high molecular weight acidic heteropolymer exopolysaccharide, is a virulent factor of E. amylovora. As reported earlier, amylovoran production in an amyR knockout mutant was about eight-fold higher than that in the wild type (WT) strain of E. amylovora. When a multicopy plasmid containing the amyR gene was introduced into the amyR mutant or WT strains, amylovoran production was strongly inhibited. Furthermore, amylovoran production was also suppressed in various amylovoran-over-producing mutants, such as grrSA containing multicopies of the amyR gene. Consistent with amylovoran production, an inverse correlation was observed between in vitro expression of amyR and that of amylovoran biosynthetic genes. However, both the amyR knockout mutant and over-expression strains showed reduced levan production, another exopolysaccharide produced by E. amylovora. Virulence assays demonstrated that while the amyR mutant was capable of inducing slightly greater disease severity than that of the WT strain, strains over-expressing the amyR gene did not incite disease on apple shoots or leaves, and only caused reduced disease on immature pear fruits. Microarray studies revealed that amylovoran biosynthesis and related membrane protein-encoding genes were highly expressed in the amyR mutant, but down-regulated in the amyR over-expression strains in vitro. Down-regulation of amylovoran biosynthesis genes in the amyR over-expression strain partially explained why over-expression of amyR led to non-pathogenic or reduced virulence in vivo. These results suggest that AmyR plays an important role in regulating exopolysaccharide production, and thus virulence in E. amylovora.

  5. Erwinia amylovora CRISPR elements provide new tools for evaluating strain diversity and for microbial source tracking.

    Science.gov (United States)

    McGhee, Gayle C; Sundin, George W

    2012-01-01

    Clustered regularly interspaced short palindromic repeats (CRISPRs) comprise a family of short DNA repeat sequences that are separated by non repetitive spacer sequences and, in combination with a suite of Cas proteins, are thought to function as an adaptive immune system against invading DNA. The number of CRISPR arrays in a bacterial chromosome is variable, and the content of each array can differ in both repeat number and in the presence or absence of specific spacers. We utilized a comparative sequence analysis of CRISPR arrays of the plant pathogen Erwinia amylovora to uncover previously unknown genetic diversity in this species. A total of 85 E. amylovora strains varying in geographic isolation (North America, Europe, New Zealand, and the Middle East), host range, plasmid content, and streptomycin sensitivity/resistance were evaluated for CRISPR array number and spacer variability. From these strains, 588 unique spacers were identified in the three CRISPR arrays present in E. amylovora, and these arrays could be categorized into 20, 17, and 2 patterns types, respectively. Analysis of the relatedness of spacer content differentiated most apple and pear strains isolated in the eastern U.S. from western U.S. strains. In addition, we identified North American strains that shared CRISPR genotypes with strains isolated on other continents. E. amylovora strains from Rubus and Indian hawthorn contained mostly unique spacers compared to apple and pear strains, while strains from loquat shared 79% of spacers with apple and pear strains. Approximately 23% of the spacers matched known sequences, with 16% targeting plasmids and 5% targeting bacteriophage. The plasmid pEU30, isolated in E. amylovora strains from the western U.S., was targeted by 55 spacers. Lastly, we used spacer patterns and content to determine that streptomycin-resistant strains of E. amylovora from Michigan were low in diversity and matched corresponding streptomycin-sensitive strains from the

  6. Molecular signature of differential virulence in natural isolates of Erwinia amylovora.

    Science.gov (United States)

    Wang, Dongping; Korban, Schuyler S; Zhao, Youfu

    2010-02-01

    ABSTRACT Erwinia amylovora, the causal agent of fire blight, is considered to be a genetically homogeneous species based on physiological, biochemical, phylogenetic, and genetic analysis. However, E. amylovora strains exhibiting differential virulence are isolated from nature. The exopolysaccharide amylovoran and type III secretion system (T3SS) are two major yet separate virulence factors in E. amylovora. The objective of this study was to investigate whether there is a correlation between E. amylovora virulence and levels of virulence gene expression. Four wild-type strains (Ea1189, Ea273, Ea110, and CFBP1430), widely used in studies of E. amylovora pathogenesis, have been analyzed and compared. E. amylovora strains Ea273 and Ea110 elicited higher severity of disease symptoms than those of Ea1189 and CFBP1430 on apple cv. Golden Delicious and G16 apple root stock plants but not on susceptible Gala plants. In addition, Ea273 and Ea110 elicited severe hypersensitive responses within shorter periods of time at lower inoculum concentrations than those of Ea1189 and CFBP1430 on tobacco plants. Further molecular analyses have revealed that amylovoran production and expression of both amylovoran (amsG) and T3SS (dspE and hrpL) genes were significantly higher in Ea273 and Ea110 than those in Ea1189 and CFBP1430. Other phenotypes such as swarming motility in these four strains also differed significantly. These results indicate that E. amylovora strains of different origin can be divided into subgroups based on molecular signatures of virulence gene expression. Therefore, these molecular signatures may be used to differentiate E. amylovora strains, which may have taxonomical and evolutionary implications.

  7. Erwinia amylovora CRISPR elements provide new tools for evaluating strain diversity and for microbial source tracking.

    Directory of Open Access Journals (Sweden)

    Gayle C McGhee

    Full Text Available Clustered regularly interspaced short palindromic repeats (CRISPRs comprise a family of short DNA repeat sequences that are separated by non repetitive spacer sequences and, in combination with a suite of Cas proteins, are thought to function as an adaptive immune system against invading DNA. The number of CRISPR arrays in a bacterial chromosome is variable, and the content of each array can differ in both repeat number and in the presence or absence of specific spacers. We utilized a comparative sequence analysis of CRISPR arrays of the plant pathogen Erwinia amylovora to uncover previously unknown genetic diversity in this species. A total of 85 E. amylovora strains varying in geographic isolation (North America, Europe, New Zealand, and the Middle East, host range, plasmid content, and streptomycin sensitivity/resistance were evaluated for CRISPR array number and spacer variability. From these strains, 588 unique spacers were identified in the three CRISPR arrays present in E. amylovora, and these arrays could be categorized into 20, 17, and 2 patterns types, respectively. Analysis of the relatedness of spacer content differentiated most apple and pear strains isolated in the eastern U.S. from western U.S. strains. In addition, we identified North American strains that shared CRISPR genotypes with strains isolated on other continents. E. amylovora strains from Rubus and Indian hawthorn contained mostly unique spacers compared to apple and pear strains, while strains from loquat shared 79% of spacers with apple and pear strains. Approximately 23% of the spacers matched known sequences, with 16% targeting plasmids and 5% targeting bacteriophage. The plasmid pEU30, isolated in E. amylovora strains from the western U.S., was targeted by 55 spacers. Lastly, we used spacer patterns and content to determine that streptomycin-resistant strains of E. amylovora from Michigan were low in diversity and matched corresponding streptomycin-sensitive strains

  8. PR genes of apple: identification and expression in response to elicitors and inoculation with Erwinia amylovora

    Directory of Open Access Journals (Sweden)

    Kim Jihyun F

    2006-10-01

    Full Text Available Abstract Background In the past decade, much work has been done to dissect the molecular basis of the defence signalling pathway in plants known as Systemic Acquired Resistance (SAR. Most of the work has been carried out in model species such as Arabidopsis, with little attention paid to woody plants. However within the range of species examined, components of the pathway seem to be highly conserved. In this study, we attempted to identify downstream components of the SAR pathway in apple to serve as markers for its activation. Results We identified three pathogenesis related (PR genes from apple, PR-2, PR-5 and PR-8, which are induced in response to inoculation with the apple pathogen, Erwinia amylovora, but they are not induced in young apple shoots by treatment with known elicitors of SAR in herbaceous plants. We also identified three PR-1-like genes from apple, PR-1a, PR-1b and PR-1c, based solely on sequence similarity to known PR-1 genes of model (intensively researched herbaceous plants. The PR-1-like genes were not induced in response to inoculation with E. amylovora or by treatment with elicitors; however, each showed a distinct pattern of expression. Conclusion Four PR genes from apple were partially characterized. PR-1a, PR-2, PR-5 and PR-8 from apple are not markers for SAR in young apple shoots. Two additional PR-1-like genes were identified through in-silico analysis of apple ESTs deposited in GenBank. PR-1a, PR-1b and PR-1c are not involved in defence response or SAR in young apple shoots; this conclusion differs from that reported previously for young apple seedlings.

  9. Genetic and virulence variability among Erwinia tracheiphila strains recovered from different cucurbit hosts.

    Science.gov (United States)

    Rojas, E Saalau; Dixon, P M; Batzer, J C; Gleason, M L

    2013-09-01

    The causal agent of cucurbit bacterial wilt, Erwinia tracheiphila, has a wide host range in the family Cucurbitaceae, including economically important crops such as muskmelon (Cucumis melo), cucumber (C. sativus), and squash (Cucurbita spp.). Genetic variability of 69 E. tracheiphila strains was investigated by repetitive-element polymerase chain reaction (rep-PCR) using BOXA1R and ERIC1-2 primers. Fingerprint profiles revealed significant variability associated with crop host; strains isolated from Cucumis spp. were clearly distinguishable from Cucurbita spp.-isolated strains regardless of geographic origin. Twelve E. tracheiphila strains isolated from muskmelon, cucumber, or summer squash were inoculated onto muskmelon and summer squash seedlings, followed by incubation in a growth chamber. Wilt symptoms were assessed over 3 weeks, strains were reisolated, and rep-PCR profiles were compared with the inoculated strains. Wilting occurred significantly faster when seedlings were inoculated with strains that originated from the same crop host genus (P<0.001). In the first run of the experiment, cucumber and muskmelon strains caused wilting on muskmelon seedlings at a median of 7.8 and 5.6 days after inoculation (dai), respectively. Summer squash seedlings wilted 18.0, 15.7, and 5.7 dai when inoculated with muskmelon-, cucumber-, and squash-origin strains, respectively. In a second run of the experiment, cucumber and muskmelon strains caused wilting on muskmelon at 7.0 and 6.9 dai, respectively, whereas summer squash seedlings wilted at 23.6, 29.0 and 9.0 dai when inoculated with muskmelon-, cucumber-, and squash-origin strains, respectively. Our results provide the first evidence of genetic diversity within E. tracheiphila and suggest that strain specificity is associated with plant host. This advance is a first step toward understanding the genetic and population structure of E. tracheiphila.

  10. Crystallization and preliminary crystallographic analysis of l-asparaginase from Erwinia carotovora

    Energy Technology Data Exchange (ETDEWEB)

    Wikman, Linnea E. K. [Turku Centre for Biotechnology, University of Turku and Åbo Akademi University, Turku 20521 (Finland); Krasotkina, Julya; Kuchumova, Anastasia; Sokolov, Nikolay N. [Institute for Biomedical Chemistry, Russian Academy of Medical Sciences, 559-B, 10 Pogodinskay St, Moscow 119121 (Russian Federation); Papageorgiou, Anastassios C., E-mail: tassos.papageorgiou@btk.fi [Turku Centre for Biotechnology, University of Turku and Åbo Akademi University, Turku 20521 (Finland)

    2005-04-01

    Er. carotovoral-asparaginase, a potential antileukaemic agent, has been crystallized. Crystals diffract to 2.6 Å using a rotating-anode source and belong to space group P2{sub 1}, with unit-cell parameters a = 78.0, b = 112.3, c = 78.7 Å, β = 101.9° and a homotetramer in the crystallographic asymmetric unit. Bacterial l-asparaginases have been used as therapeutic agents in the treatment of acute childhood lymphoblastic leukaemia for over 30 y. However, their use is limited owing to the glutaminase activity of the administered enzymes, which results in serious side effects. In contrast, l-asparaginase from Erwinia carotovora exhibits low glutaminase activity at physiological concentrations of l-asparagine and l-glutamine in the blood. Recombinant Er. carotovoral-asparaginase was crystallized in the presence of l-glutamate by the hanging-drop vapour-diffusion method using 10 mg ml{sup −1} purified enzyme, 16–18%(w/v) PEG 3350 and 0.2 M NaF. X-ray diffraction data were collected to 2.6 Å at 293 K using an in-house rotating-anode generator. The crystals belong to the monoclinic P2{sub 1} space group, with unit-cell parameters a = 78.0, b = 112.3, c = 78.7 Å, β = 101.9° and a homotetramer in the crystallographic asymmetric unit. A molecular-replacement solution has been found and refinement is currently in progress. The crystal structure may provide leads towards protein-engineering efforts aimed at safer asparaginase administration in leukaemia treatment.

  11. Identification of gene-specific markers for resistance to Erwinia amylovora (fire blight) in Malus (apple) by a functional genomics approach

    Science.gov (United States)

    Fire blight, caused by Erwinia amylovora (Ea), is a destructive disease of apple (Malus), pear (Pyrus) and other plants in the rose family (Rosaceae). 650 expressed sequence tags (ESTs) associated with fire blight were identified from Ea-challenged apple leaf tissue by suppression subtractive hybrid...

  12. Molecular and physiological properties of bacteriophages from North America and Germany affecting the fire blight pathogen Erwinia amylovora.

    Science.gov (United States)

    Müller, Ina; Lurz, Rudi; Kube, Michael; Quedenau, Claudia; Jelkmann, Wilhelm; Geider, Klaus

    2011-11-01

    For possible control of fire blight affecting apple and pear trees, we characterized Erwinia amylovora phages from North America and Germany. The genome size determined by electron microscopy (EM) was confirmed by sequence data and major coat proteins were identified from gel bands by mass spectroscopy. By their morphology from EM data, φEa1h and φEa100 were assigned to the Podoviridae and φEa104 and φEa116 to the Myoviridae. Host ranges were essentially confined to E. amylovora, strains of the species Erwinia pyrifoliae, E. billingiae and even Pantoea stewartii were partially sensitive. The phages φEa1h and φEa100 were dependent on the amylovoran capsule of E. amylovora, φEa104 and φEa116 were not. The Myoviridae efficiently lysed their hosts and protected apple flowers significantly better than the Podoviridae against E. amylovora and should be preferred in biocontrol experiments. We have also isolated and partially characterized E. amylovora phages from apple orchards in Germany. They belong to the Podoviridae or Myoviridae with a host range similar to the phages isolated in North America. In EM measurements, the genome sizes of the Podoviridae were smaller than the genomes of the Myoviridae from North America and from Germany, which differed from each other in corresponding nucleotide sequences.

  13. Gene-for-gene relationship in the host-pathogen system Malus × robusta 5-Erwinia amylovora.

    Science.gov (United States)

    Vogt, Isabelle; Wöhner, Thomas; Richter, Klaus; Flachowsky, Henryk; Sundin, George W; Wensing, Annette; Savory, Elizabeth A; Geider, Klaus; Day, Brad; Hanke, Magda-Viola; Peil, Andreas

    2013-03-01

    Fire blight is a destructive bacterial disease caused by Erwinia amylovora affecting plants in the family Rosaceae, including apple. Host resistance to fire blight is present mainly in accessions of Malus spp. and is thought to be quantitative in this pathosystem. In this study we analyzed the importance of the E. amylovora effector avrRpt2(EA) , a homolog of Pseudomonas syringae avrRpt2, for resistance of Malus × robusta 5 (Mr5). The deletion mutant E. amylovora Ea1189ΔavrRpt2(EA) was able to overcome the fire blight resistance of Mr5. One single nucleotide polymorphism (SNP), resulting in an exchange of cysteine to serine in the encoded protein, was detected in avrRpt2(EA) of several Erwinia strains differing in virulence to Mr5. E. amylovora strains encoding serine (S-allele) were able to overcome resistance of Mr5, whereas strains encoding cysteine (C-allele) were not. Allele specificity was also observed in a coexpression assay with Arabidopsis thaliana RIN4 in Nicotiana benthamiana. A homolog of RIN4 has been detected and isolated in Mr5. These results suggest a system similar to the interaction of RPS2 from A. thaliana and AvrRpt2 from P. syringae with RIN4 as guard. Our data are suggestive of a gene-for-gene relationship for the host-pathogen system Mr5 and E. amylovora.

  14. Production of isomaltulose obtained by Erwinia sp. cells submitted to different treatments and immobilized in calcium alginate

    Directory of Open Access Journals (Sweden)

    Haroldo Yukio Kawaguti

    2011-03-01

    Full Text Available In recent decades, there has been an increase in the studies of isomaltulose obtainment, due to its physicochemical properties and physiological health benefits. These properties, which include low cariogenicity, low glycemic index and greater stability, allow the use of this sweetener as a substitute for sucrose in foods; besides the fact that it can be converted to isomalt, a dietary non-cariogenic sugar alcohol used in pharmaceuticals as well as in the food industry. Isomaltulose (6-O-α-D-glucopyronosyl-1-6-D-fructofuranose is a disaccharide reducer obtained by the enzymatic conversion of sucrose - the α-glucosyltransferase enzyme. Different treatments were performed for the preparation of whole cells; lysed cells; and crude enzyme extract of Erwinia sp. D12 strain immobilized in calcium alginate. The packed bed column of granules, containing Erwinia sp. cells sonicated and immobilized in calcium alginate (CSI, reached a maximum conversion of 53-59% sucrose into isomaltulose and it presented activity for 480 hours. The converted syrup was purified and the isomaltulose crystallization was performed through the lowering of temperature. The isomaltulose crystals presented purity of 96.5%.

  15. hexA of Erwinia carotovora ssp. carotovora strain Ecc71 negatively regulates production of RpoS and rsmB RNA, a global regulator of extracellular proteins, plant virulence and the quorum-sensing signal, N-(3-oxohexanoyl)-L-homoserine lactone.

    Science.gov (United States)

    Mukherjee, A; Cui, Y; Ma, W; Liu, Y; Chatterjee, A K

    2000-04-01

    The soft-rotting bacterium, Erwinia carotovora ssp. carotovora (E. c. carotovora), produces an array of extracellular enzymes (= exoenzymes), including pectate lyase (Pel), polygalacturonase (Peh), cellulase (Cel) and protease (Prt), as well as HarpinEcc, the elicitor of hypersensitive reaction (HR). The production of these exoenzymes and HarpinEcc responds to plant products and the quorum-sensing signal [N-(3-oxohexanoyl)-L-homoserine lactone; OHL] and is subject to both transcriptional and post-transcriptional regulation. hexA of E. c. carotovora strain Ecc71 (hereafter hexA71), like that of another E. c. carotovora strain, negatively controls the production of exoenzymes, OHL and virulence in E. c. carotovora strain Ecc71. In addition to exoenzymes, HexA71 negatively regulates the expression of hrpNEcc, the structural gene for HarpinEcc. Exoenzyme overproduction is abolished by OHL deficiency in a HexA- and Ohll- double mutant, indicating that HexA and OHL are components of a common regulatory pathway controlling exoenzyme production. HexA71 negatively affects RpoS, as the levels of this alternative sigma factor are higher in the HexA- mutant than in the HexA+ strain. However, a HexA- and RpoS double mutant produces higher levels of exoenzymes and transcripts of pel-1, peh-1 and celVgenes than the HexA- and RpoS+ parent. Thus, the elevated levels of RpoS protein in the HexA- mutant do not account for exoenzyme overproduction. The following evidence associates for the first time the phenotypic changes in the HexA mutant to overproduction of rsmB RNA, a global regulator of exoenzymes, HarpinEcc, OHL and secondary metabolites. Analyses of rsmB transcripts and expression of an rsmB-lacZoperon fusion in E. c. carotovora strain Ecc71 revealed that HexA71 negatively regulates transcription of rsmB. Multiple copies of hexA71+ DNA suppress various phenotypes, including exoenzyme production in E. c. carotovora strain Ecc71, and concomitantly inhibit the production of rsm

  16. Functional Characterization of Twin-arginine Translocation Gene (tatC) from Erwinia amylovora%梨火疫病菌(Erwinia amylovora)双精氨酸运输系统基因(tatC)的功能分析

    Institute of Scientific and Technical Information of China (English)

    于洋洋; 刘倩倩; 徐恩丽; 胡白石

    2011-01-01

    Erwinia amylovom causes fire blight on many plants of the Rosaceae family such as apple and pear. The Twin-arginine translocation (Tat) pathway plays a crucial role in transporting the proteins which are related to virulence. In this study, a tatC disruption mutant EaAtatC was successfully constructed by homologous recombination as well as the complement strain EaAtatC (pME-tatC). The results showed that the tatC mutant EaAtafC displayed the reduced virulence, extracellular polysaccharide production, chemotaxis, motility, flagella assembly and growth in vitro. However, compared to wild type strain NCPPB1665 (Eal665), the induction of hypersensitive response in nonhost tobacco, biofilm synthesis and cellulose production of the tatC mutant EaAtalC didn't show significant difference. These findings demonstrate that tatC gene in Erwinia amylovora is involved in growth, and motility and play an important role in virulence.%梨火疫病菌(Erwinia amylovora)可引起梨、苹果等蔷薇科植物的火疫病.双精氨酸运输系统(Tat)与致病相关蛋白的转运有特定的联系.本研究在梨火疫病菌全基因组中发现了同源基因,通过同源重组的方法,构建了梨火疫病菌的tatC基因突变体以及互补子.研究结果表明,tatC基因影响着梨火疫病菌的致病性、胞外多糖、鞭毛运动、游动性、趋化性、生长情况等多种生物学特性.然而,Ea△tatC仍能引起烟草过敏性反应,并且在胞外纤维素和生物膜的生成方面与野生型菌株相比没有明显差异.说明梨火疫病菌tatC基因对病菌的生长、游动性以及致病性方面具有关键作用.

  17. 野菊花乙醚提取物的理化性质及抗氧化能力%Physicochemical Characterization and Antioxidant Activity of Ether Extract from Flos chrysanthemi indici

    Institute of Scientific and Technical Information of China (English)

    申海进; 郭巧生; 房海灵

    2012-01-01

    Physicochemical proprieties and antioxidant activity of ether extract from Flos chrysanthemi indici were investigated in this study.The yield of dried ether extract from crude materials(dry weight) was 2.15%.The content of carotenoids in dried extract was(165.51 ± 1.42) mg/g.The ether extract was added to four kinds of edible oil including peanut oil,soybean oil,sunflower oil and corn oil to evaluate its antioxidant activity.Our results indicated that the ether extract could effectively inhibit lipid oxidation and displayed a potential antioxidant activity in a dose-dependent manner.Therefore,this ether extract may be considered as a potential antioxidant agent.%研究野菊花乙醚提取物的基本理化性质及其抗氧化特性。结果表明:野菊花乙醚提取物中类胡萝卜素含量为(165.51±1.42)mg/g,β-胡萝卜素含量为(2537.96±11.57)μg/g;抗氧化实验显示提取物均能够有效阻止花生油、大豆油、葵花籽油、玉米油的脂质快速氧化,且具有剂量效应关系。野菊花乙醚提取物是一种潜在的油脂氧化保护剂,具备一定的抗氧化能力。

  18. Research progress on chemical constituents and pharmacological activities of Chrysanthemi Flos%杭白菊化学成分和药理活性的研究进展

    Institute of Scientific and Technical Information of China (English)

    钟爱娇; 姜哲; 李雪征; 李宁

    2014-01-01

    杭白菊具有较高的药用价值,民间应用广泛,是卫生部首批批准的药食同源的道地药材之一。杭白菊主要含有黄酮、挥发油、三萜、甾体、酚类、多糖及微量元素等多种成分,具有心血管保护、降血脂、血糖、血压、抗氧化、抗癌、神经保护、肝保护等多种药理活性。为了进一步开发和合理利用该药用植物资源,综述了近年来国内外对杭白菊的化学成分和药理活性的研究进展。%Chrysanthemi Flosis widely used in folk with high medicinal value. It was firstly approved as one of the medicinal and edible herbals by Ministry of Public Health. Phytochemical studies revealed that it contained flavonoids, volatile oils, triterpenes, sterols, phenols, polysaccharides, trace elements and other physiologically active substances. Furthermore, it showed many pharmaceutical effects, such as cardiovascular protective and hypolipidemic, hyperglycemic and hypotesive activities, antioxidative, anti-cancer, neuroprotective effects, hepatoprotective effects, and so on. To further develop and rationally use this plant resource, this article summarizes the chemical constituents and pharmacological activities ofChrysanthemi Flosin recent years.

  19. Isolation of a Bacterium Strain Degraded Agar

    Institute of Scientific and Technical Information of China (English)

    2010-01-01

    One in 58 strains of bacteria isolated from the compost showed clear colonies after a few days of growth on the plates containing medium made of only agar and water.Water suspension contained only agar (2 and 8g·L -1 ) with two controls (normal saline,LB medium) was inoculated with the bacterium BR5-1 to see whether there was an increasement of the alive bacteria concentration after 48 h of the growth.The results showed that there was a significant rising of the alive bacteria concentration in the agar susp...

  20. Swimming Efficiency of Bacterium Escherichia Coli

    CERN Document Server

    Chattopadhyay, S; Wu, X L; Yeung, C; Chattopadhyay, Suddhashil; Moldovan, Radu; Yeung, Chuck

    2005-01-01

    We use in vivo measurements of swimming bacteria in an optical trap to determine fundamental properties of bacterial propulsion. In particular, we determine the propulsion matrix, which relates the angular velocity of the flagellum to the torques and forces propelling the bacterium. From the propulsion matrix dynamical properties such as forces, torques, swimming speed and power can be obtained from measurements of the angular velocity of the motor. We find significant heterogeneities among different individuals even though all bacteria started from a single colony. The propulsive efficiency, defined as the ratio of the propulsive power output to the rotary power input provided by the motors, is found to be 0.2%.

  1. Characterization of Streptomycin Resistance in Isolates of Erwinia amylovora in California.

    Science.gov (United States)

    Förster, Helga; McGhee, Gayle C; Sundin, George W; Adaskaveg, James E

    2015-10-01

    In surveys from 2006 to 2014, streptomycin resistance in Erwinia amylovora from pear-growing areas in California declined from very high incidence in 2006 and 2007 to very low incidence in 2013 and 2014. The majority of resistant strains were designated as moderately resistant-low (MR-L), and were almost exclusively found in Sacramento County, whereas highly resistant (HR) strains were only recovered in Sutter-Yuba and San Joaquin counties. Resistance of HR strains was associated with a mutation in codon 43 of the chromosomal rpsL gene that results in a change from lysine to arginine, the same mutation that was originally reported for resistant strains from California in the mid-1970s. MR-L strains were found to harbor the strA-strB streptomycin resistance genes on transposon Tn5393a. This transposon lacks insertion sequence IS1133 that provides a promoter for efficient expression of strA-strB, resulting in lower minimum inhibitory concentrations of MR-L strains compared with those from other locations that harbor strA-strB on Tn5393::IS1133. In contrast to previously described plasmid-mediated resistance where Tn5393 is inserted in pEa34, or pEA29, Tn5393a in MR-L strains was located on plasmid pEU30. This plasmid was first described in E. amylovora from the western United States but was not associated with streptomycin resistance determinants previously. We hypothesize that Tn5393a was introduced into an E. amylovora strain carrying pEU30 and transposed into that plasmid. This hypothesis was supported by clustered regularly interspaced short palindromic repeat (CRISPR) sequence analysis that showed that two MR-L strains share the same CRISPR1 pattern as a streptomycin-sensitive strain. With current low resistance levels in California growing regions, streptomycin could be successfully used again, but applications per season should be limited and the antibiotic should be mixed and rotated with different modes of action.

  2. Erwinia carotovora elicitors and Botrytis cinerea activate defense responses in Physcomitrella patens

    Directory of Open Access Journals (Sweden)

    Bentancor Marcel

    2007-10-01

    Full Text Available Abstract Background Vascular plants respond to pathogens by activating a diverse array of defense mechanisms. Studies with these plants have provided a wealth of information on pathogen recognition, signal transduction and the activation of defense responses. However, very little is known about the infection and defense responses of the bryophyte, Physcomitrella patens, to well-studied phytopathogens. The purpose of this study was to determine: i whether two representative broad host range pathogens, Erwinia carotovora ssp. carotovora (E.c. carotovora and Botrytis cinerea (B. cinerea, could infect Physcomitrella, and ii whether B. cinerea, elicitors of a harpin (HrpN producing E.c. carotovora strain (SCC1 or a HrpN-negative strain (SCC3193, could cause disease symptoms and induce defense responses in Physcomitrella. Results B. cinerea and E.c. carotovora were found to readily infect Physcomitrella gametophytic tissues and cause disease symptoms. Treatments with B. cinerea spores or cell-free culture filtrates from E.c. carotovoraSCC1 (CF(SCC1, resulted in disease development with severe maceration of Physcomitrella tissues, while CF(SCC3193 produced only mild maceration. Although increased cell death was observed with either the CFs or B. cinerea, the occurrence of cytoplasmic shrinkage was only visible in Evans blue stained protonemal cells treated with CF(SCC1 or inoculated with B. cinerea. Most cells showing cytoplasmic shrinkage accumulated autofluorescent compounds and brown chloroplasts were evident in a high proportion of these cells. CF treatments and B. cinerea inoculation induced the expression of the defense-related genes: PR-1, PAL, CHS and LOX. Conclusion B. cinerea and E.c. carotovora elicitors induce a defense response in Physcomitrella, as evidenced by enhanced expression of conserved plant defense-related genes. Since cytoplasmic shrinkage is the most common morphological change observed in plant PCD, and that harpins and B

  3. Biodegradation of heavy oils by halophilic bacterium

    Institute of Scientific and Technical Information of China (English)

    Ruixia Hao; Anhuai Lu

    2009-01-01

    A halophilic bacterial strain TM-1 was isolated from the reservoir of the Shengli oil field in East China. Strain TM-1, which was found to be able to degrade crude oils, is a gram-positive non-motile bacterium with a coccus shape that can grow at temperatures of up to 58 ℃ and in 18% NaCl solution. Depending on the culture conditions, the organism may occur in tetrads. In addition, strain TM-1 pro-duced acid from glucose without gas formation and was catalase-negative. Furthermore, strain TM-I was found to be a facultative aer-obe capable of growth under anaerobic conditions. Moreover, it produced butylated hydroxytoluene, 1,2-benzenedicarboxylic acid-bis ester and dibutyl phthalate and could use different organic substrates. Laboratory studies indicated that strain TM-1 affected different heavy oils by degrading various components and by changing the chemical properties of the oils. In addition, growth of the bacterium in heavy oils resulted in the loss of aromatic hydrocarbons, resins and asphaltenes, and enrichment with light hydrocarbons and an overall redistribution of these hydrocarbons.

  4. Eliminação da produção de bacteriocinas em Erwinia e Pseudomonas fitopatogênicas Elimination of bacteriocin production in the phytopathogenic bacteria Erwinia and Pseudomonas

    Directory of Open Access Journals (Sweden)

    C.M.R. de Biagi

    1995-04-01

    Full Text Available Duas linhagens de bactérias fitopatogênicas do gênero Pseudomonas e duas do gênero Erwinia produtoras de bacteriocinas, foram submetidas a diferentes tratamentos visando estimar a estabilidade desse caráter. Houve eliminação da produção de bacteriocinas após tratamento com duas concentrações de brometo de etídio e a porcentagem de eliminação variou com o hospedeiro e com a concentração da droga. Não houve eliminação em temperaturas elevadas e também a preservação da linhagem por três anos em laboratório não causou perda do caráter. A eliminação da produção de bacteriocinas com brometo de etídio sugere que os genes envolvidos tenham localização plasmidiana.Four strains, two from each of the plant pathogenic bacteria Pseudomonas and Erwinia, all producing bacteriocins, were submitted to different treatments in order to evaluate the persistance of bacteriocin production. After ethidium bromide treatment elimination of the bacteriocin production was achieved and the percentage of loss varied with the host strain and drug concentration. Elimination was not detected after treatment with high temperatures, above the normal ones. Also preservation of the strains for three years, in laboratory, did not cause elimination of the character. The curing of bacteriocin production after ethidium bromide treatment suggests that the involved genes are located in plasmids.

  5. A Sequential Statistical Approach towards an Optimized Production of a Broad Spectrum Bacteriocin Substance from a Soil Bacterium Bacillus sp. YAS 1 Strain

    Directory of Open Access Journals (Sweden)

    Amira M. Embaby

    2014-01-01

    Full Text Available Bacteriocins, ribosomally synthesized antimicrobial peptides, display potential applications in agriculture, medicine, and industry. The present study highlights integral statistical optimization and partial characterization of a bacteriocin substance from a soil bacterium taxonomically affiliated as Bacillus sp. YAS 1 after biochemical and molecular identifications. A sequential statistical approach (Plackett-Burman and Box-Behnken was employed to optimize bacteriocin (BAC YAS 1 production. Using optimal levels of three key determinants (yeast extract (0.48% (w/v, incubation time (62 hrs, and agitation speed (207 rpm in peptone yeast beef based production medium resulted in 1.6-fold enhancement in BAC YAS 1 level (470 AU/mL arbitrary units against Erwinia amylovora. BAC YAS 1 showed activity over a wide range of pH (1–13 and temperature (45–80°C. A wide spectrum antimicrobial activity of BAC YAS 1 against the human pathogens (Clostridium perfringens, Staphylococcus epidermidis, Campylobacter jejuni, Enterobacter aerogenes, Enterococcus sp., Proteus sp., Klebsiella sp., and Salmonella typhimurium, the plant pathogen (E. amylovora, and the food spoiler (Listeria innocua was demonstrated. On top and above, BAC YAS 1 showed no antimicrobial activity towards lactic acid bacteria (Lactobacillus bulgaricus, L. casei, L. lactis, and L. reuteri. Promising characteristics of BAC YAS 1 prompt its commercialization for efficient utilization in several industries.

  6. A sequential statistical approach towards an optimized production of a broad spectrum bacteriocin substance from a soil bacterium Bacillus sp. YAS 1 strain.

    Science.gov (United States)

    Embaby, Amira M; Heshmat, Yasmin; Hussein, Ahmed; Marey, Heba S

    2014-01-01

    Bacteriocins, ribosomally synthesized antimicrobial peptides, display potential applications in agriculture, medicine, and industry. The present study highlights integral statistical optimization and partial characterization of a bacteriocin substance from a soil bacterium taxonomically affiliated as Bacillus sp. YAS 1 after biochemical and molecular identifications. A sequential statistical approach (Plackett-Burman and Box-Behnken) was employed to optimize bacteriocin (BAC YAS 1) production. Using optimal levels of three key determinants (yeast extract (0.48% (w/v), incubation time (62 hrs), and agitation speed (207 rpm)) in peptone yeast beef based production medium resulted in 1.6-fold enhancement in BAC YAS 1 level (470 AU/mL arbitrary units against Erwinia amylovora). BAC YAS 1 showed activity over a wide range of pH (1-13) and temperature (45-80 °C). A wide spectrum antimicrobial activity of BAC YAS 1 against the human pathogens (Clostridium perfringens, Staphylococcus epidermidis, Campylobacter jejuni, Enterobacter aerogenes, Enterococcus sp., Proteus sp., Klebsiella sp., and Salmonella typhimurium), the plant pathogen (E. amylovora), and the food spoiler (Listeria innocua) was demonstrated. On top and above, BAC YAS 1 showed no antimicrobial activity towards lactic acid bacteria (Lactobacillus bulgaricus, L. casei, L. lactis, and L. reuteri). Promising characteristics of BAC YAS 1 prompt its commercialization for efficient utilization in several industries.

  7. Genome comparison of the epiphytic bacteria Erwinia billingiae and E. tasmaniensis with the pear pathogen E. pyrifoliae

    Directory of Open Access Journals (Sweden)

    Kuhl Heiner

    2010-06-01

    Full Text Available Abstract Background The genus Erwinia includes plant-associated pathogenic and non-pathogenic Enterobacteria. Important pathogens such as Erwinia amylovora, the causative agent of fire blight and E. pyrifoliae causing bacterial shoot blight of pear in Asia belong to this genus. The species E. tasmaniensis and E. billingiae are epiphytic bacteria and may represent antagonists for biocontrol of fire blight. The presence of genes that are putatively involved in virulence in E. amylovora and E. pyrifoliae is of special interest for these species in consequence. Results Here we provide the complete genome sequences of the pathogenic E. pyrifoliae strain Ep1/96 with a size of 4.1 Mb and of the non-pathogenic species E. billingiae strain Eb661 with a size of 5.4 Mb, de novo determined by conventional Sanger sequencing and next generation sequencing techniques. Genome comparison reveals large inversions resulting from homologous recombination events. Furthermore, comparison of deduced proteins highlights a relation of E. billingiae strain Eb661 to E. tasmaniensis strain Et1/99 and a distance to E. pyrifoliae for the overall gene content as well as for the presence of encoded proteins representing virulence factors for the pathogenic species. Pathogenicity of E. pyrifoliae is supposed to have evolved by accumulation of potential virulence factors. E. pyrifoliae carries factors for type III secretion and cell invasion. Other genes described as virulence factors for E. amylovora are involved in the production of exopolysaccharides, the utilization of plant metabolites such as sorbitol and sucrose. Some virulence-associated genes of the pathogenic species are present in E. tasmaniensis but mostly absent in E. billingiae. Conclusion The data of the genome analyses correspond to the pathogenic lifestyle of E. pyrifoliae and underlines the epiphytic localization of E. tasmaniensis and E. billingiae as a saprophyte.

  8. Complete genome sequence of the fire blight pathogen Erwinia pyrifoliae DSM 12163T and comparative genomic insights into plant pathogenicity

    Directory of Open Access Journals (Sweden)

    Frey Jürg E

    2010-01-01

    Full Text Available Abstract Background Erwinia pyrifoliae is a newly described necrotrophic pathogen, which causes fire blight on Asian (Nashi pear and is geographically restricted to Eastern Asia. Relatively little is known about its genetics compared to the closely related main fire blight pathogen E. amylovora. Results The genome of the type strain of E. pyrifoliae strain DSM 12163T, was sequenced using both 454 and Solexa pyrosequencing and annotated. The genome contains a circular chromosome of 4.026 Mb and four small plasmids. Based on their respective role in virulence in E. amylovora or related organisms, we identified several putative virulence factors, including type III and type VI secretion systems and their effectors, flagellar genes, sorbitol metabolism, iron uptake determinants, and quorum-sensing components. A deletion in the rpoS gene covering the most conserved region of the protein was identified which may contribute to the difference in virulence/host-range compared to E. amylovora. Comparative genomics with the pome fruit epiphyte Erwinia tasmaniensis Et1/99 showed that both species are overall highly similar, although specific differences were identified, for example the presence of some phage gene-containing regions and a high number of putative genomic islands containing transposases in the E. pyrifoliae DSM 12163T genome. Conclusions The E. pyrifoliae genome is an important addition to the published genome of E. tasmaniensis and the unfinished genome of E. amylovora providing a foundation for re-sequencing additional strains that may shed light on the evolution of the host-range and virulence/pathogenicity of this important group of plant-associated bacteria.

  9. Genetic characterization of the HrpL regulon of the fire blight pathogen Erwinia amylovora reveals novel virulence factors.

    Science.gov (United States)

    McNally, R Ryan; Toth, Ian K; Cock, Peter J A; Pritchard, Leighton; Hedley, Pete E; Morris, Jenny A; Zhao, Youfu; Sundin, George W

    2012-02-01

    The bacterial pathogen Erwinia amylovora is the causal agent of fire blight, an economically significant disease of apple and pear. Disease initiation by E. amylovora requires the translocation of effector proteins into host cells via the hypersensitive response and pathogenicity (hrp) type III secretion system (T3SS). The alternative sigma factor HrpL positively regulates the transcription of structural and translocated components of the T3SS via hrp promoter elements. To characterize genome-wide HrpL-dependent gene expression in E. amylovora Ea1189, wild-type and Ea1189ΔhrpL strains were cultured in hrp-inducing minimal medium, and total RNA was compared using a custom microarray designed to represent the annotated genes of E. amylovora ATCC 49946. The results revealed 24 genes differentially regulated in Ea1189ΔhrpL relative to Ea1189 with fold-change expression ratios greater than 1.5; of these, 19 genes exhibited decreased transcript abundance and five genes showed increased transcript abundance relative to Ea1189. To expand our understanding of the HrpL regulon and to elucidate direct versus indirect HrpL-mediated effects on gene expression, the genome of E. amylovora ATCC 49946 was examined in silico using a hidden Markov model assembled from known Erwinia spp. hrp promoters. This technique identified 15 putative type III novel hrp promoters, seven of which were validated with quantitative polymerase chain reaction based on expression analyses. It was found that HrpL-regulated genes encode all known components of the hrp T3SS, as well as five putative type III effectors. Eight genes displayed apparent indirect HrpL regulation, suggesting that the HrpL regulon is connected to downstream signalling networks. The construction of deletion mutants of three novel HrpL-regulated genes resulted in the identification of additional virulence factors as well as mutants displaying abnormal motility and biofilm phenotypes.

  10. Gene Cloning and Characterization of luxR Transcriptional Regulator from Erwinia amylovora%梨火疫病菌luxR转录调节因子的克隆与功能分析

    Institute of Scientific and Technical Information of China (English)

    吴惠秋; 徐恩丽; 于洋洋; 田艳丽; 钱国良; 刘凤权; 胡白石

    2013-01-01

    Erwinia amylovora is a necrogenic phytopathogenic bacterium,the causal agent of fire blight in pear(Pyrus sorotina),apple(Malus domestica) and other members of the Rosaceae family,causing serious economic losses.LuxR transcriptional regulator is a key player in quorum sensing and plays important physiological and biochemical roles.In this paper,one of luxR homologous gene was amplified by PCR from E.amylovora.The mutant Ea△luxR was successfully constucted by homologous recombination,and the function studied preliminarily.The experimental results showed that,compared to the wild-type strain,the polyene macrolide biosynthesis,anti-oxidative stress,growth,and pathogenicity of mutant Ea△luxR declined.However,Ea△luxR still caused tobacco(Nicotiana tabacum) hypersensitive response,and there was no significant difference in antibiotic tolerance level and biofilm formation comparing to wild-type strain.These findings demonstrated that luxR gene in E.amylovora is involved in polyene macrolide biosynthesis,growth,and anti-oxidative stress and plays an important role in virulence.In addition,our results provid molecular evidence for in-depth understanding of the quorum sensing system of E.amylovora.%梨火疫病菌(Erwinia amylovora)是一种具有毁灭性的植物病原细菌,能够侵染梨、苹果和其他蔷薇科植物引起火疫病,造成严重的经济损失.luxR家族调控因子具有重要的生理生化作用,是细菌群体感应机制中研究较多的一类重要的转录调节基因.本研究采用PCR技术从梨火疫病菌中扩增出一个luxR同源基因,命名为EaluxR,应用同源重组的方法构建了突变株(Ea△luxR),并对其功能进行初步研究.实验结果表明,Ea△luxR与野生型菌株相比产生的多烯大环内酯类代谢产物在抗氧化压力和生长情况方面有明显差异,对梨幼苗及幼果的致病能力下降;但是Ea△luxR仍能引起烟草过敏性反应,并且在抗生素耐受水

  11. Diffusion of magnetotactic bacterium in rotating magnetic field

    Energy Technology Data Exchange (ETDEWEB)

    Cebers, A., E-mail: aceb@tesla.sal.l [Department of Physics, University of Latvia, Zellu 8, Ri-bar ga, LV-1002 (Latvia)

    2011-02-15

    Swimming trajectory of a magnetotactic bacterium in a rotating magnetic field is a circle. Random reversals of the direction of the bacterium motion induces a random walk of the curvature center of the trajectory. In assumption of the distribution of the switching events according to the Poisson process the diffusion coefficient is calculated in dependence on the frequency of the rotating field and the characteristic time between the switching events. It is confirmed by the numerical simulation of the random walk of the bacterium in the rotating magnetic field. - Research highlights: Random switching of the flagella leads to diffusion of a bacterium in the field. Mean square displacement of the curvature center is proportional to time. Diffusion coefficient depends on the period of a rotating field. At zero frequency diffusion coefficient is the same as for a tumbling bacterium.

  12. Fluctuation-Enhanced Sensing of Bacterium Odors

    CERN Document Server

    Chang, Hung-Chih; King, Maria D; Kwan, Chiman

    2009-01-01

    The goal of this paper is to explore the possibility to detect and identify bacteria by sensing their odor via fluctuation-enhanced sensing with commercial Taguchi sensors. The fluctuations of the electrical resistance during exposure to different bacterial odors, Escherichia coli and anthrax-surrogate Bacillus subtilis, have been measured and analyzed. In the present study, the simplest method, the measurement and analysis of power density spectra was used. The sensors were run in the normal heated and the sampling-and-hold working modes, respectively. The results indicate that Taguchi sensors used in these fluctuation-enhanced modes are effective tools of bacterium detection and identification even when they are utilizing only the power density spectrum of the stochastic sensor signal.

  13. The chemical formula of a magnetotactic bacterium.

    Science.gov (United States)

    Naresh, Mohit; Das, Sayoni; Mishra, Prashant; Mittal, Aditya

    2012-05-01

    Elucidation of the chemical logic of life is one of the grand challenges in biology, and essential to the progress of the upcoming field of synthetic biology. Treatment of microbial cells explicitly as a "chemical" species in controlled reaction (growth) environments has allowed fascinating discoveries of elemental formulae of a few species that have guided the modern views on compositions of a living cell. Application of mass and energy balances on living cells has proved to be useful in modeling of bioengineering systems, particularly in deriving optimized media compositions for growing microorganisms to maximize yields of desired bio-derived products by regulating intra-cellular metabolic networks. In this work, application of elemental mass balance during growth of Magnetospirillum gryphiswaldense in bioreactors has resulted in the discovery of the chemical formula of the magnetotactic bacterium. By developing a stoichiometric equation characterizing the formation of a magnetotactic bacterial cell, coupled with rigorous experimental measurements and robust calculations, we report the elemental formula of M. gryphiswaldense cell as CH(2.06)O(0.13)N(0.28)Fe(1.74×10(-3)). Remarkably, we find that iron metabolism during growth of this magnetotactic bacterium is much more correlated individually with carbon and nitrogen, compared to carbon and nitrogen with each other, indicating that iron serves more as a nutrient during bacterial growth rather than just a mineral. Magnetotactic bacteria have not only invoked some interest in the field of astrobiology for the last two decades, but are also prokaryotes having the unique ability of synthesizing membrane bound intracellular organelles. Our findings on these unique prokaryotes are a strong addition to the limited repertoire, of elemental compositions of living cells, aimed at exploring the chemical logic of life.

  14. Effect of concentration and substrate flow rate on isomaltulose production from sucrose by Erwinia sp. cells immobilized in calcium-alginate using packed bed reactor.

    Science.gov (United States)

    Kawaguti, Haroldo Yukio; Harumi Sato, Hélia

    2010-09-01

    Isomaltulose was obtained from sucrose solution by immobilized cells of Erwinia sp. D12 using a batch and a continuous process. Parameters for sucrose conversion into isomaltulose were evaluated using both experimental design and response surface methodology. Erwinia sp. D12 cells were immobilized in different alginates, and the influence of substrate flow rate and concentration parameters to produce isomaltulose from sucrose were observed. Response surface methodology demonstrated that packed bed columns containing cells immobilized in low-viscosity sodium alginate (250 cP) presented a mean isomaltulose conversion rate of 47%. In a continuous process, both sucrose substrate concentration and substrate flow rate parameters had a significant effect (p < 0.05) and influenced the conversion of sucrose into isomaltulose. Higher conversion rates of sucrose into isomaltulose, from 53-75% were obtained using 75 g of immobilized cells at a substrate flow rate of 0.6 mL/min.

  15. The outer membrane protein TolC is required for phytoalexin resistance and virulence of the fire blight pathogen Erwinia amylovora

    OpenAIRE

    2009-01-01

    Summary Erwinia amylovora causes fire blight on several plant species such as apple and pear, which produce diverse phytoalexins as defence mechanisms. An evolutionary successful pathogen thus must develop resistance mechanisms towards these toxic compounds. The E. amylovora outer membrane protein, TolC, might mediate phytoalexin resistance through its interaction with the multidrug efflux pump, AcrAB. To prove this, a tolC mutant and an acrB/tolC double mutant were constructed. The minimal i...

  16. Comparative analysis of the Hrp pathogenicity island of Rubus- and Spiraeoideae-infecting Erwinia amylovora strains identifies the IT region as a remnant of an integrative conjugative element.

    Science.gov (United States)

    Mann, Rachel A; Blom, Jochen; Bühlmann, Andreas; Plummer, Kim M; Beer, Steven V; Luck, Joanne E; Goesmann, Alexander; Frey, Jürg E; Rodoni, Brendan C; Duffy, Brion; Smits, Theo H M

    2012-08-01

    The Hrp pathogenicity island (hrpPAI) of Erwinia amylovora not only encodes a type III secretion system (T3SS) and other genes required for pathogenesis on host plants, but also includes the so-called island transfer (IT) region, a region that originates from an integrative conjugative element (ICE). Comparative genomic analysis of the IT regions of two Spiraeoideae- and three Rubus-infecting strains revealed that the regions in Spiraeoideae-infecting strains were syntenic and highly conserved in length and genetic information, but that the IT regions of the Rubus-infecting strains varied in gene content and length, showing a mosaic structure. None of the ICEs in E. amylovora strains were complete, as conserved ICE genes and the left border were missing, probably due to reductive genome evolution. Comparison of the hrpPAI region of E. amylovora strains to syntenic regions from other Erwinia spp. indicates that the hrpPAI and the IT regions are the result of several insertion and deletion events that have occurred within the ICE. It also suggests that the T3SS was present in a common ancestor of the pathoadapted Erwinia spp. and that insertion and deletion events in the IT region occurred during speciation.

  17. Experimental evolution of aging in a bacterium

    Directory of Open Access Journals (Sweden)

    Stearns Stephen C

    2007-07-01

    Full Text Available Abstract Background Aging refers to a decline in reproduction and survival with increasing age. According to evolutionary theory, aging evolves because selection late in life is weak and mutations exist whose deleterious effects manifest only late in life. Whether the assumptions behind this theory are fulfilled in all organisms, and whether all organisms age, has not been clear. We tested the generality of this theory by experimental evolution with Caulobacter crescentus, a bacterium whose asymmetric division allows mother and daughter to be distinguished. Results We evolved three populations for 2000 generations in the laboratory under conditions where selection was strong early in life, but very weak later in life. All populations evolved faster growth rates, mostly by decreasing the age at first division. Evolutionary changes in aging were inconsistent. The predominant response was the unexpected evolution of slower aging, revealing the limits of theoretical predictions if mutations have unanticipated phenotypic effects. However, we also observed the spread of a mutation causing earlier aging of mothers whose negative effect was reset in the daughters. Conclusion Our results confirm that late-acting deleterious mutations do occur in bacteria and that they can invade populations when selection late in life is weak. They suggest that very few organisms – perhaps none- can avoid the accumulation of such mutations over evolutionary time, and thus that aging is probably a fundamental property of all cellular organisms.

  18. Pantoea agglomerans: a marvelous bacterium of evil and good.Part I. Deleterious effects: Dust-borne endotoxins and allergens – focus on cotton dust

    Directory of Open Access Journals (Sweden)

    Jacek Dutkiewicz

    2015-12-01

    Full Text Available The ubiquitous Gram-negative bacterium Pantoea agglomerans (synonyms: Enterobacter agglomerans, Erwinia herbicola is known both as an epiphytic microbe developing on the surface of plants and as an endophytic organism living inside the plants. The bacterium occurs also abundantly in plant and animal products, in the body of arthropods and other animals, in water, soil, dust and air, and occasionally in humans. From the human viewpoint, the role of this organism is ambiguous, both deleterious and beneficial: on one side it causes disorders in people exposed to inhalation of organic dusts and diseases of crops, and on the other side it produces substances effective in the treatment of cancer and other diseases of humans and animals, suppresses the development of various plant pathogens, promotes plant growth, and appears as a potentially efficient biofertilizer and bioremediator. P. agglomerans was identified as a predominant bacterium on cotton plant grown all over the world, usually as an epiphyte, rarely as pathogen. It is particularly numerous on cotton bract after senescence. During processing of cotton in mills, bacteria and their products are released with cotton dust into air and are inhaled by workers, causing respiratory and general disorders, usually defined as byssinosis. The most adverse substance is endotoxin, a heteropolymer macromolecule present in the outermost part of the cell wall, consisting of lipopolysaccharide (LPS as a major constituent, phospholipids and protein. The numerous experiments carried out in last quarter of XXth century on laboratory animals and human volunteers supported a convincing evidence that the inhaled endotoxin produced by P. agglomerans causes numerous pathologic effects similar to those elicited by cotton dust, such as influx of free lung cells into airways and activation of alveolar macrophages which secrete mediators (prostaglandins, platelet-activating factor, interleukin-1, tumor necrosis factor

  19. Erwinia herbicola冰核活性蛋白的分离、电泳分析鉴定%Differential centrifugal isolation and identification of electrophoresis analysis to ice-nucleating activity protein of Erwinia herbicola

    Institute of Scientific and Technical Information of China (English)

    陈庆森; 张晓玲; 阎亚丽; 刘剑虹; 庞广昌

    2002-01-01

    Erwinia herbicola(A25)菌株的冰核活性蛋白的分离纯化及电泳进行研究.主要方法①按双温培养的方法,获得了较高的生物量积累和强冰核活性的诱导表达.②实验采用渗透压冲击法破碎细菌细胞,破碎率达98.67%.③通过差速离心法获取不同冰核活性蛋白组分,测定各组分的冰核活性和SDS-PAGE电泳图谱分析.建立了冰核活性因子的高冰核活性与离心力之间的关系;利用SDS-PAGE还建立了具冰核活性蛋白的分子量的大小与冰核活性蛋白组分之间的关系.证实了具高冰核活性蛋白质最小结构单位约为26.0kD.

  20. Taxonomic characterization of the cellulose-degrading bacterium NCIB 10462

    Energy Technology Data Exchange (ETDEWEB)

    Dees, C.; Ringleberg, D.; Scott, T.C. [Oak Ridge National Lab., TN (United States); Phelps, T. [Univ. of Tennessee, Knoxville, TN (United States)

    1994-06-01

    The gram negative cellulase-producing bacterium NCIB 10462 has been previously named Pseudomonas fluorescens subsp. or var. cellulosa. Since there is renewed interest in cellulose-degrading bacteria for use in bioconversion of cellulose to chemical feed stocks and fuels, we re-examined the characteristics of this microorganism to determine its proper taxonomic characterization and to further define it`s true metabolic potential. Metabolic and physical characterization of NCIB 10462 revealed that this was an alkalophilic, non-fermentative, gram negative, oxidase positive, motile, cellulose-degrading bacterium. The aerobic substrate utilization profile of this bacterium was found to have few characteristics consistent with a classification of P. fluorescens with a very low probability match with the genus Sphingomonas. Total lipid analysis did not reveal that any sphingolipid bases are produced by this bacterium. NCIB 10462 was found to grow best aerobically but also grows well in complex media under reducing conditions. NCIB 10462 grew slowly under full anaerobic conditions on complex media but growth on cellulosic media was found only under aerobic conditions. Total fatty acid analysis (MIDI) of NCIB 10462 failed to group this bacterium with a known pseudomonas species. However, fatty acid analysis of the bacteria when grown at temperatures below 37{degrees}C suggest that the organism is a pseudomonad. Since a predominant characteristic of this bacterium is it`s ability to degrade cellulose, we suggest it be called Pseudomonas cellulosa.

  1. Pangenome Evolution in the Marine Bacterium Alteromonas.

    Science.gov (United States)

    López-Pérez, Mario; Rodriguez-Valera, Francisco

    2016-06-03

    We have examined a collection of the free-living marine bacterium Alteromonas genomes with cores diverging in average nucleotide identities ranging from 99.98% to 73.35%, i.e., from microbes that can be considered members of a natural clone (like in a clinical epidemiological outbreak) to borderline genus level. The genomes were largely syntenic allowing a precise delimitation of the core and flexible regions in each. The core was 1.4 Mb (ca. 30% of the typical strain genome size). Recombination rates along the core were high among strains belonging to the same species (37.7-83.7% of all nucleotide polymorphisms) but they decreased sharply between species (18.9-5.1%). Regarding the flexible genome, its main expansion occurred within the boundaries of the species, i.e., strains of the same species already have a large and diverse flexible genome. Flexible regions occupy mostly fixed genomic locations. Four large genomic islands are involved in the synthesis of strain-specific glycosydic receptors that we have called glycotypes. These genomic regions are exchanged by homologous recombination within and between species and there is evidence for their import from distant taxonomic units (other genera within the family). In addition, several hotspots for integration of gene cassettes by illegitimate recombination are distributed throughout the genome. They code for features that give each clone specific properties to interact with their ecological niche and must flow fast throughout the whole genus as they are found, with nearly identical sequences, in different species. Models for the generation of this genomic diversity involving phage predation are discussed.

  2. Diversity, evolution, and functionality of clustered regularly interspaced short palindromic repeat (CRISPR) regions in the fire blight pathogen Erwinia amylovora.

    Science.gov (United States)

    Rezzonico, Fabio; Smits, Theo H M; Duffy, Brion

    2011-06-01

    The clustered regularly interspaced short palindromic repeat (CRISPR)/Cas system confers acquired heritable immunity against mobile nucleic acid elements in prokaryotes, limiting phage infection and horizontal gene transfer of plasmids. In CRISPR arrays, characteristic repeats are interspersed with similarly sized nonrepetitive spacers derived from transmissible genetic elements and acquired when the cell is challenged with foreign DNA. New spacers are added sequentially and the number and type of CRISPR units can differ among strains, providing a record of phage/plasmid exposure within a species and giving a valuable typing tool. The aim of this work was to investigate CRISPR diversity in the highly homogeneous species Erwinia amylovora, the causal agent of fire blight. A total of 18 CRISPR genotypes were defined within a collection of 37 cosmopolitan strains. Strains from Spiraeoideae plants clustered in three major groups: groups II and III were composed exclusively of bacteria originating from the United States, whereas group I generally contained strains of more recent dissemination obtained in Europe, New Zealand, and the Middle East. Strains from Rosoideae and Indian hawthorn (Rhaphiolepis indica) clustered separately and displayed a higher intrinsic diversity than that of isolates from Spiraeoideae plants. Reciprocal exclusion was generally observed between plasmid content and cognate spacer sequences, supporting the role of the CRISPR/Cas system in protecting against foreign DNA elements. However, in several group III strains, retention of plasmid pEU30 is inconsistent with a functional CRISPR/Cas system.

  3. Expression of the dspA/E gene of Erwinia amylovora in non-host plant Arabidopsis thaliana

    Directory of Open Access Journals (Sweden)

    Hasan Murat Aksoy

    2017-01-01

    Full Text Available In the Erwinia amylovora genome, the hrp gene cluster containing the dspA/E/EB/F operon plays a crucial role in mediating the pathogenicity and the hypersensitive response (HR in the host plant. The role of the dspA/E gene derived from E. amylovora was investigated by monitoring the expression of the β-glucuronidase (GUS reporter system in transgenic Arabidopsis thaliana cv. Pri-Gus seedlings. A mutant ΔdspA/E strain of E. amylovora was generated to contain a deletion of the dspA/E gene for the purpose of this study. Two-week-old seedlings of GUS transgenic Arabidopsis were vacuum-infiltrated with the wild-type and the mutant (ΔdspA/E E. amylovora strains. The Arabidopsis seedlings were fixed and stained for GUS activity after 3–5 days following infiltration. The appearance of dense spots with blue staining on the Arabidopsis leaves indicated the typical characteristic of GUS activity. This observation indicated that the wild-type E. amylovora strain had induced a successful and efficient infection on the A. thaliana Pri-Gus leaves. In contrast, there was no visible GUS expression on leaf tissues which were inoculated with the ΔdspA/E mutant E. amylovora strain. These results indicate that the dspA/E gene is required by the bacterial cells to induce HR in non-host plants.

  4. Small-molecule inhibitors suppress the expression of both type III secretion and amylovoran biosynthesis genes in Erwinia amylovora.

    Science.gov (United States)

    Yang, Fan; Korban, Schuyler S; Pusey, P Lawrence; Elofsson, Michael; Sundin, George W; Zhao, Youfu

    2014-01-01

    The type III secretion system (T3SS) and exopolysaccharide (EPS) amylovoran are two essential pathogenicity factors in Erwinia amylovora, the causal agent of the serious bacterial disease fire blight. In this study, small molecules that inhibit T3SS gene expression in E. amylovora under hrp (hypersensitive response and pathogenicity)-inducing conditions were identified and characterized using green fluorescent protein (GFP) as a reporter. These compounds belong to salicylidene acylhydrazides and also inhibit amylovoran production. Microarray analysis of E. amylovora treated with compounds 3 and 9 identified a total of 588 significantly differentially expressed genes. Among them, 95 and 78 genes were activated and suppressed by both compounds, respectively, when compared with the dimethylsulphoxide (DMSO) control. The expression of the majority of T3SS genes in E. amylovora, including hrpL and the avrRpt2 effector gene, was suppressed by both compounds. Compound 3 also suppressed the expression of amylovoran precursor and biosynthesis genes. However, both compounds induced significantly the expression of glycogen biosynthesis genes and siderophore biosynthesis, regulatory and transport genes. Furthermore, many membrane, lipoprotein and exported protein-encoding genes were also activated by both compounds. Similar expression patterns were observed for compounds 1, 2 and 4. Using crab apple flower as a model, compound 3 was capable of reducing disease development in pistils. These results suggest a common inhibition mechanism shared by salicylidene acylhydrazides and indicate that small-molecule inhibitors that disable T3SS function could be explored to control fire blight disease.

  5. A novel plasmid pEA68 of Erwinia amylovora and the description of a new family of plasmids.

    Science.gov (United States)

    Ismail, Emadeldeen; Blom, Jochen; Bultreys, Alain; Ivanović, Milan; Obradović, Aleksa; van Doorn, Joop; Bergsma-Vlami, Maria; Maes, Martine; Willems, Anne; Duffy, Brion; Stockwell, Virginia O; Smits, Theo H M; Puławska, Joanna

    2014-12-01

    Recent genome analysis of Erwinia amylovora, the causal agent of fire blight disease on Rosaceae, has shown that the chromosome is highly conserved among strains and that plasmids are the principal source of genomic diversity. A new circular plasmid, pEA68, was found in E. amylovora strain 692 (LMG 28361), isolated in Poland from Sorbus (mountain ash) with fire blight symptoms. Annotation of the 68,763-bp IncFIIa-type plasmid revealed that it contains 79 predicted CDS, among which two operons (tra, pil) are associated with mobility. The plasmid is maintained stably in E. amylovora and does not possess genes associated with antibiotic resistance or known virulence genes. Curing E. amylovora strain 692 of pEA68 did not influence its virulence in apple shoots nor amylovoran synthesis. Of 488 strains of E. amylovora from seventeen countries, pEA68 was only found in two additional strains from Belgium. Although the spread of pEA68 is currently limited to Europe, pEA68 comprises, together with pEA72 and pEA78 both found in North America, a new plasmid family that spans two continents.

  6. Comparative genomics of 12 strains of Erwinia amylovora identifies a pan-genome with a large conserved core.

    Science.gov (United States)

    Mann, Rachel A; Smits, Theo H M; Bühlmann, Andreas; Blom, Jochen; Goesmann, Alexander; Frey, Jürg E; Plummer, Kim M; Beer, Steven V; Luck, Joanne; Duffy, Brion; Rodoni, Brendan

    2013-01-01

    The plant pathogen Erwinia amylovora can be divided into two host-specific groupings; strains infecting a broad range of hosts within the Rosaceae subfamily Spiraeoideae (e.g., Malus, Pyrus, Crataegus, Sorbus) and strains infecting Rubus (raspberries and blackberries). Comparative genomic analysis of 12 strains representing distinct populations (e.g., geographic, temporal, host origin) of E. amylovora was used to describe the pan-genome of this major pathogen. The pan-genome contains 5751 coding sequences and is highly conserved relative to other phytopathogenic bacteria comprising on average 89% conserved, core genes. The chromosomes of Spiraeoideae-infecting strains were highly homogeneous, while greater genetic diversity was observed between Spiraeoideae- and Rubus-infecting strains (and among individual Rubus-infecting strains), the majority of which was attributed to variable genomic islands. Based on genomic distance scores and phylogenetic analysis, the Rubus-infecting strain ATCC BAA-2158 was genetically more closely related to the Spiraeoideae-infecting strains of E. amylovora than it was to the other Rubus-infecting strains. Analysis of the accessory genomes of Spiraeoideae- and Rubus-infecting strains has identified putative host-specific determinants including variation in the effector protein HopX1(Ea) and a putative secondary metabolite pathway only present in Rubus-infecting strains.

  7. Fire blight disease reactome: RNA-seq transcriptional profile of apple host plant defense responses to Erwinia amylovora pathogen infection.

    Science.gov (United States)

    Kamber, Tim; Buchmann, Jan P; Pothier, Joël F; Smits, Theo H M; Wicker, Thomas; Duffy, Brion

    2016-02-17

    The molecular basis of resistance and susceptibility of host plants to fire blight, a major disease threat to pome fruit production globally, is largely unknown. RNA-sequencing data from challenged and mock-inoculated flowers were analyzed to assess the susceptible response of apple to the fire blight pathogen Erwinia amylovora. In presence of the pathogen 1,080 transcripts were differentially expressed at 48 h post inoculation. These included putative disease resistance, stress, pathogen related, general metabolic, and phytohormone related genes. Reads, mapped to regions on the apple genome where no genes were assigned, were used to identify potential novel genes and open reading frames. To identify transcripts specifically expressed in response to E. amylovora, RT-PCRs were conducted and compared to the expression patterns of the fire blight biocontrol agent Pantoea vagans strain C9-1, another apple pathogen Pseudomonas syringae pv. papulans, and mock inoculated apple flowers. This led to the identification of a peroxidase superfamily gene that was lower expressed in response to E. amylovora suggesting a potential role in the susceptibility response. Overall, this study provides the first transcriptional profile by RNA-seq of the host plant during fire blight disease and insights into the response of susceptible apple plants to E. amylovora.

  8. The luxS gene is involved in AI-2 production, pathogenicity, and some phenotypes in Erwinia amylovora.

    Science.gov (United States)

    Gao, Yan; Song, Junxian; Hu, Baishi; Zhang, Lei; Liu, Qianqian; Liu, Fengquan

    2009-01-01

    Erwinia amylovora causes fire blight of apple, pear, and other members of the Rosaceae family. The enzyme LuxS catalyzes the last step in the production of autoinducer-2 (AI-2), a molecule implicated with quorum sensing in many bacterial species. It is now well recognized that LuxS also plays a central role in sulfur metabolism and in the activated methyl cycle, which is responsible for the generation of S-adenosyl-L-methionine. A research paper has reported that luxS is not involved with quorum sensing in Er. amylovora, but in our study, Er. amylovora strain NCPPB1665 (Ea1665) produced luxS-dependent extracellular AI-2 activity. Additionally, the maximal AI-2 activity occurred during late-exponential and early-stationary growth phases and diminished during the stationary phase. The luxS mutant of Ea1665 was constructed, and the phenotypes of a defined luxS mutant have been characterized. Inactivation of luxS in Ea1665 impaired motility, extracellular polysaccharide (EPS) production, and tolerance for hydrogen peroxide, and reduced virulence on pear leaves.

  9. Comparative genomics of 12 strains of Erwinia amylovora identifies a pan-genome with a large conserved core.

    Directory of Open Access Journals (Sweden)

    Rachel A Mann

    Full Text Available The plant pathogen Erwinia amylovora can be divided into two host-specific groupings; strains infecting a broad range of hosts within the Rosaceae subfamily Spiraeoideae (e.g., Malus, Pyrus, Crataegus, Sorbus and strains infecting Rubus (raspberries and blackberries. Comparative genomic analysis of 12 strains representing distinct populations (e.g., geographic, temporal, host origin of E. amylovora was used to describe the pan-genome of this major pathogen. The pan-genome contains 5751 coding sequences and is highly conserved relative to other phytopathogenic bacteria comprising on average 89% conserved, core genes. The chromosomes of Spiraeoideae-infecting strains were highly homogeneous, while greater genetic diversity was observed between Spiraeoideae- and Rubus-infecting strains (and among individual Rubus-infecting strains, the majority of which was attributed to variable genomic islands. Based on genomic distance scores and phylogenetic analysis, the Rubus-infecting strain ATCC BAA-2158 was genetically more closely related to the Spiraeoideae-infecting strains of E. amylovora than it was to the other Rubus-infecting strains. Analysis of the accessory genomes of Spiraeoideae- and Rubus-infecting strains has identified putative host-specific determinants including variation in the effector protein HopX1(Ea and a putative secondary metabolite pathway only present in Rubus-infecting strains.

  10. Phenylalanine in the pore of the Erwinia ligand-gated ion channel modulates picrotoxinin potency but not receptor function.

    Science.gov (United States)

    Thompson, Andrew J; Alqazzaz, Mona; Price, Kerry L; Weston, David A; Lummis, Sarah C R

    2014-10-01

    The Erwinia ligand-gated ion channel (ELIC) is a bacterial homologue of eukaryotic Cys-loop ligand-gated ion channels. This protein has the potential to be a useful model for Cys-loop receptors but is unusual in that it has an aromatic residue (Phe) facing into the pore, leading to some predictions that this protein is incapable of ion flux. Subsequent studies have shown this is not the case, so here we probe the role of this residue by examining the function of the ELIC in cases in which the Phe has been substituted with a range of alternative amino acids, expressed in Xenopus oocytes and functionally examined. Most of the mutations have little effect on the GABA EC50, but the potency of the weak pore-blocking antagonist picrotoxinin at F16'A-, F16'D-, F16'S-, and F16'T-containing receptors was increased to levels comparable with those of Cys-loop receptors, suggesting that this antagonist can enter the pore only when residue 16' is small. T6'S has no effect on picrotoxinin potency when expressed alone but abolishes the increased potency when combined with F16'S, indicating that the inhibitor binds at position 6', as in Cys-loop receptors, if it can enter the pore. Overall, the data support the proposal that the ELIC pore is a good model for Cys-loop receptor pores if the role of F16' is taken into consideration.

  11. HOPX1 Ea (Eop3) in Erwinia Amylovora functions as an avirulence gene in apple and is regulated by HRPL

    Science.gov (United States)

    Fire blight is a devastating disease of rosaceous plants caused by the Gram-negative bacterium E. amylovora. This pathogen delivers virulence proteins into host cells utilizing the Type-Three Secretion System (T3SS). Expression of the T3SS and associated substrates are activated by the alternative s...

  12. Seca dos ponteiros da goiabeira causada por Erwinia psidii: níveis de incidência e aspectos epidemiológicos Guava bacterial blight due to Erwinia psidii: incidence levels and epidemiological aspects

    Directory of Open Access Journals (Sweden)

    Abi Soares Anjos Marques

    2007-01-01

    Full Text Available Um dos fatores limitantes ao cultivo da goiabeira no Brasil é a 'seca dos ponteiros', causada por Erwinia psidii, presente nas regiões Sudeste e Centro-Oeste, onde se concentram grandes áreas produtoras. Considerando a pequena disponibilidade de informações sobre a epidemiologia e níveis de incidência dessa bacteriose, este estudo teve como objetivos: confirmar a distribuição e verificar a dispersão da seca dos ponteiros da goiabeira no Distrito Federal; investigar o efeito da temperatura sobre a multiplicação in vitro de E. psidii; desenvolver um teste de patogenicidade prático e eficiente e avaliar a sobrevivência in vitro da bactéria em diferentes substratos. A doença foi identificada em 56% das propriedades produtoras avaliadas no DF, com 81,9% de correlação entre a presença de sintomas e o diagnóstico laboratorial. A melhor faixa de temperatura para multiplicação de E. psidii foi de 24 a 33 ºC, e a bactéria permaneceu viável por até 120 dias em suspensão em água. A inoculação da bactéria em folhas ou hastes destacadas levou ao aparecimento de sintomas a partir do sétimo dia e mostrou-se eficiente como um teste rápido para se avaliar a patogenicidade de isolados.A major disease that affects guava is 'bacterial blight', caused by Erwinia psidii, which has been reported in Southeastern and Central Regions of Brazil where the major producing areas are located. Considering the lack of information on epidemiology and incidence levels of this disease, the objectives of this study were to confirm the presence and to verify the spread of the disease in Distrito Federal (DF; to determine optimal temperature for in vitro multiplication of E. psidii; to develop a simple and effective method for pathogenicity testing and to evaluate in vitro bacterial survival on different substrates. The disease was detected in 56% of producing orchards evaluated in DF, with a correlation of 81, 9% between presence of symptoms and

  13. Characterization of a cfr-Carrying Plasmid from Porcine Escherichia coli That Closely Resembles Plasmid pEA3 from the Plant Pathogen Erwinia amylovora.

    Science.gov (United States)

    Zhang, Rongmin; Sun, Bin; Wang, Yang; Lei, Lei; Schwarz, Stefan; Wu, Congming

    2015-11-02

    The multiresistance gene cfr was found in two porcine Escherichia coli isolates, one harboring it on the conjugative 33,885-bp plasmid pFSEC-01, the other harboring it in the chromosomal DNA. Sequence analysis of pFSEC-01 revealed that a 6,769-bp fragment containing the cfr gene bracketed by two IS26 elements was inserted into a plasmid closely related to pEA3 from the plant pathogen Erwinia amylovora, suggesting that pFSEC-01 may be transferred between different bacterial genera of both animal and plant origin.

  14. Deep sequencing revealed genome-wide single-nucleotide polymorphism and plasmid content of Erwinia amylovora strains isolated in Middle Atlas, Morocco.

    Science.gov (United States)

    Hannou, Najat; Mondy, Samuel; Planamente, Sara; Moumni, Mohieddine; Llop, Pablo; López, María; Manceau, Charles; Barny, Marie-Anne; Faure, Denis

    2013-10-01

    Erwinia amylovora causes economic losses that affect pear and apple production in Morocco. Here, we report comparative genomics of four Moroccan E. amylovora strains with the European strain CFBP1430 and North-American strain ATCC49946. Analysis of single nucleotide polymorphisms (SNPs) revealed genetic homogeneity of Moroccan's strains and their proximity to the European strain CFBP1430. Moreover, the collected sequences allowed the assembly of a 65 kpb plasmid, which is highly similar to the plasmid pEI70 harbored by several European E. amylovora isolates. This plasmid was found in 33% of the 40 E. amylovora strains collected from several host plants in 2009 and 2010 in Morocco.

  15. Antibacterial activity of moss extracts on Erwinia carotovora%藓类提取物对白菜软腐病菌的抑菌活性

    Institute of Scientific and Technical Information of China (English)

    程辉彩; 赵建成

    2006-01-01

    以欧文氏菌(Erwinia carotovora)为供试菌,对24种藓类乙醇提取物进行抑菌活性筛选.结果表明,有15种藓类提取物对欧文氏菌表现出不同程度抑菌作用,并将有较强抑菌作用的提取物对欧文氏菌进行了最低抑菌浓度MIC测定.

  16. Alternative sigma factor RpoN and its modulation protein YhbH are indispensable for Erwinia amylovora virulence.

    Science.gov (United States)

    Ancona, Veronica; Li, Wenting; Zhao, Youfu

    2014-01-01

    In Erwinia amylovora, ECF (extracytoplasmic functions) alternative sigma factor HrpL regulates the transcription of hrp (hypersensitive response and pathogenicity)-type III secretion system (T3SS) genes by binding to a consensus sequence known as the hrp box in hrp gene promoters. In turn, the expression of hrpL has been proposed to be positively controlled by alternative sigma factor 54 (σ(54)) (RpoN) and HrpS, a member of the σ(54) enhancer-binding proteins (EBPs). However, the function of RpoN has not been characterized genetically in E. amylovora. In this study, we investigated the role of RpoN, a nitrogen limitation sigma factor, and its modulation protein YhbH, a novel ribosome-associated protein, in E. amylovora virulence. Our results showed that mutations in hrpS, hrpL, rpoN and yhbH, but not yfiA and rmf3, resulted in a nonpathogenic phenotype on immature pear fruits and apple shoots. Consistently, the expression of T3SS genes, including hrpL, dspE, hrpN and hrpA, was barely detected in hrpS, hrpL, rpoN and yhbH mutants. These mutants were also not capable of eliciting a hypersensitive response (HR) on tobacco; however, the overexpression of hrpL using an inducible promoter rescued the HR-eliciting abilities of these mutants. These results suggest that a sigma factor cascade exists in the regulatory networks of E. amylovora and regulates important virulence factors. On the basis of this study and previously reported data, a model is proposed for the regulation of T3SS in E. amylovora.

  17. Mutation of the Erwinia amylovora argD gene causes arginine auxotrophy, nonpathogenicity in apples, and reduced virulence in pears.

    Science.gov (United States)

    Ramos, Laura S; Lehman, Brian L; Peter, Kari A; McNellis, Timothy W

    2014-11-01

    Fire blight is caused by Erwinia amylovora and is the most destructive bacterial disease of apples and pears worldwide. In this study, we found that E. amylovora argD(1000)::Tn5, an argD Tn5 transposon mutant that has the Tn5 transposon inserted after nucleotide 999 in the argD gene-coding region, was an arginine auxotroph that did not cause fire blight in apple and had reduced virulence in immature pear fruits. The E. amylovora argD gene encodes a predicted N-acetylornithine aminotransferase enzyme, which is involved in the production of the amino acid arginine. A plasmid-borne copy of the wild-type argD gene complemented both the nonpathogenic and the arginine auxotrophic phenotypes of the argD(1000)::Tn5 mutant. However, even when mixed with virulent E. amylovora cells and inoculated onto immature apple fruit, the argD(1000)::Tn5 mutant still failed to grow, while the virulent strain grew and caused disease. Furthermore, the pCR2.1-argD complementation plasmid was stably maintained in the argD(1000)::Tn5 mutant growing in host tissues without any antibiotic selection. Therefore, the pCR2.1-argD complementation plasmid could be useful for the expression of genes, markers, and reporters in E. amylovora growing in planta, without concern about losing the plasmid over time. The ArgD protein cannot be considered an E. amylovora virulence factor because the argD(1000)::Tn5 mutant was auxotrophic and had a primary metabolism defect. Nevertheless, these results are informative about the parasitic nature of the fire blight disease interaction, since they indicate that E. amylovora cannot obtain sufficient arginine from apple and pear fruit tissues or from apple vegetative tissues, either at the beginning of the infection process or after the infection has progressed to an advanced state.

  18. Determination of Quantities of Host Protein after Infection with Erwinia amylovora of Apple, Pear And Quince Cultivars

    Directory of Open Access Journals (Sweden)

    Şerife Çetin

    2014-10-01

    Full Text Available Fire blight disease caused by Erwinia amylovora is a destructive bacterial pathogen mainly on pears, apples and quinces from Rosaceae family. In this study, it was aimed determination of total protein amounts in different apple cultivars (Braeburn, Fuji, Gala and Golden, pear cultivars (Santa Maria and Williams and quince cultivars (Eşme and Ekmek in the infections of two virulent E. amylovora strains (Ea234-1 and Ea240-3 according as the time. It was taken leaf samples after leaf inoculation with E. amylovora (108 CFU ml-1 at 24th, 36th and 72nd hours. For verification of the infections, re-isolations were made from bacteria inoculated plants and the agent was identified as E. amylovora by biochemical, physiological and molecular tests. In determining the amounts of total protein and in the SDS-PAGE analyses were used Bradford and Laemmli methods, respectively, and absorbance values of protein extracts derived from the leaf samples taken, were obtained at 595 nm wavelength. According to the findings obtained; after infection of E. amylovora in the apple varieties comparing to controls, total protein concentrations at 24th hours increased and a decrease in the amount of 36th to 72nd hours and Braeburn has the highest protein content was determined. In the pear varieties, while total protein concentrations at 24th and 36th hours increased, a decrease in the amount of 72nd hour, and Santa Maria variety has the highest protein content was detected. In the quince varieties, total protein concentrations at 72th hour increased and Eşme variety has the highest protein content was identified. As a result of SDS-PAGE analysis, protein fractions which have different molecular weights were obtained. The protein bands were defined approximately 55-70 kDa and 35-55 kDa molecule weight on apple and quince varieties, respectively and also approx. 55-70 kDa in pear varieties.

  19. Selection of a biocontrol agent based on a potential mechanism of action: degradation of nicotinic acid, a growth factor essential for Erwinia amylovora.

    Science.gov (United States)

    Paternoster, Thomas; Défago, Geneviève; Duffy, Brion; Gessler, Cesare; Pertot, Ilaria

    2010-12-01

    This work describes a medium-based screening method for selecting microbial biocontrol agents against Erwinia amylovora based on the degradation of a specific growth factor. Erwinia amylovora, the causal agent of the devastating fire blight disease, requires nicotinic acid or nicotinamide as an essential growth factor. Potential biocontrol agents are either selected for antimicrobial production in plate or directly on immature pears or apple blossoms. In this work, we have attempted to streamline the selection of a new potential biocontrol agent with a lower risk of non-target effects by isolation based on the ability to degrade nicotinic acid in vitro, using therefore few plant materials. A total of 735 bacteria and 1237 yeast were isolated from apple blossoms and pre-screened for nicotinic acid-degradation. Pseudomonas rhizosphaerae strain JAN was able to degrade both nicotinic acid and nicotinamide. Mutants deficient in this ability were constructed. JAN, but not the mutants, controlled E. amylovora on pear slices. On detached apple blossoms, JAN colonized apple hypanthia and strongly suppressed E. amylovora growth. Under greenhouse conditions, JAN was more effective in controlling blossom blight than P. fluorescens A506, a commercial biocontrol agent of fire blight unable to degrade nicotinic acid and nicotinamide.

  20. The Pseudomonas aeruginosa antimetabolite L-2-amino-4-methoxy-trans-3-butenoic acid inhibits growth of Erwinia amylovora and acts as a seed germination-arrest factor.

    Science.gov (United States)

    Lee, Xiaoyun; Azevedo, Mark D; Armstrong, Donald J; Banowetz, Gary M; Reimmann, Cornelia

    2013-02-01

    The Pseudomonas aeruginosa antimetabolite L-2-amino-4-methoxy-trans-3-butenoic acid (AMB) shares biological activities with 4-formylaminooxyvinylglycine, a related molecule produced by Pseudomonas fluorescens WH6. We found that culture filtrates of a P. aeruginosa strain overproducing AMB weakly interfered with seed germination of the grassy weed Poa annua and strongly inhibited growth of Erwinia amylovora, the causal agent of the devastating orchard crop disease known as fire blight. AMB was active against a 4-formylaminooxyvinylglycine-resistant isolate of E. amylovora, suggesting that the molecular targets of the two oxyvinylglycines in Erwinia do not, or not entirely, overlap. The AMB biosynthesis and transport genes were shown to be organized in two separate transcriptional units, ambA and ambBCDE, which were successfully expressed from IPTG-inducible tac promoters in the heterologous host P. fluorescens CHA0. Engineered AMB production enabled this model biocontrol strain to become inhibitory against E. amylovora and to weakly interfere with the germination of several graminaceous seeds. We conclude that AMB production requires no additional genes besides ambABCDE and we speculate that their expression in marketed fire blight biocontrol strains could potentially contribute to disease control.

  1. The genome of the Erwinia amylovora phage PhiEaH1 reveals greater diversity and broadens the applicability of phages for the treatment of fire blight.

    Science.gov (United States)

    Meczker, Katalin; Dömötör, Dóra; Vass, János; Rákhely, Gábor; Schneider, György; Kovács, Tamás

    2014-01-01

    The enterobacterium Erwinia amylovora is the causal agent of fire blight. This study presents the analysis of the complete genome of phage PhiEaH1, isolated from the soil surrounding an E. amylovora-infected apple tree in Hungary. Its genome is 218 kb in size, containing 244 ORFs. PhiEaH1 is the second E. amylovora infecting phage from the Siphoviridae family whose complete genome sequence was determined. Beside PhiEaH2, PhiEaH1 is the other active component of Erwiphage, the first bacteriophage-based pesticide on the market against E. amylovora. Comparative genome analysis in this study has revealed that PhiEaH1 not only differs from the 10 formerly sequenced E. amylovora bacteriophages belonging to other phage families, but also from PhiEaH2. Sequencing of more Siphoviridae phage genomes might reveal further diversity, providing opportunities for the development of even more effective biological control agents, phage cocktails against Erwinia fire blight disease of commercial fruit crops.

  2. Analysis of conductance responses during depolymerization of pectate by soft rot Erwinia spp. and other pectolytic bacteria isolated from potato tubers.

    Science.gov (United States)

    Fraaije, B A; Bosveld, M; Van den Bulk, R W; Rombouts, F M

    1997-07-01

    Different bacteria isolated from potato tubers were screened for their pectolytic properties by examining pitting in polypectate agar, recording conductance responses in polypectate medium and performing potato tuber soft rot tests. For bacteria found positive in conductimetry, the role of polygalacturonase (PG) and pectate lyase (PL) in the generation of conductance changes in a polygalacturonic acid (PGA) medium was further analysed using enzyme activity staining after gel electrophoresis and high-performance anion exchange chromatography. The extent of the conductance changes during depolymerization of PGA was dependent on the amounts of galacturonate monomers and oligomers accumulated in the medium. In comparison with an unidentified saprophyte and a Klebsiella strain, both mainly having PL activity, soft rot Erwinia spp. rapidly produced larger conductance responses, due to a combined action of multiple forms of PG and PL. The responses of Erwinia spp. were initially associated with the accumulation of large amounts of monomers and saturated dimers to heptamers, due to PG activity. Subsequently, as well as monomers and saturated dimers, large amounts of unsaturated dimers were also detected, due to PL activity. The role of PG as an important conductimetric factor was also demonstrated for a pectinase preparation derived from Aspergillus niger. Besides detection, automated conductimetric assays in pectate media may also be useful for monitoring of pectolytic activity in pectinase preparations and for screening of pectolytic activity of microorganisms under different media and growth conditions.

  3. Genome of a mosquito-killing bacterium decoded

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    @@ Researchers with the CAS Wuhan Institute of Virology (WHIOV) recently completed the genome sequencing of a mosquitocidal bacterium Bacillus shaericus C3-41. The feat, first of its kind in China, is expected to further promote the bio-control studies of mosquitoes.

  4. The physiology of the filamentous bacterium Microthrix parvicella

    NARCIS (Netherlands)

    Slijkhuis, H.

    1983-01-01

    A study has been made of the physiology of Microthrix parvicella. This filamentous bacterium often causes poor settleability of activated sludge in oxidation ditches supplied with domestic sewage. The organism was found to utilize only long chain fatty acids (preferably in esterified form) as carbon

  5. Domain Modeling: NP_057326.2 [SAHG[Archive

    Lifescience Database Archive (English)

    Full Text Available NP_057326.2 chr4 Crystal Structure Analysis of Pectate Lyase PelI from Erwinia chry...santhemi p3b4na_ chr4/NP_057326.2/NP_057326.2_holo_15-261.pdb swppa 16C,17S,18S,33V,34T,35T,40I,46E,47S,49Q,

  6. Domain Modeling: NP_000164.4 [SAHG[Archive

    Lifescience Database Archive (English)

    Full Text Available NP_000164.4 chr17 Crystal Structure Analysis of Pectate Lyase PelI from Erwinia chr...ysanthemi p3b4na_ chr17/NP_000164.4/NP_000164.4_holo_319-627.pdb swppa 320T,321T,322P,337Q,338M,339P,347E,35

  7. Resistance to Erwina spp. in potato (Solanum tuberosum L.).

    NARCIS (Netherlands)

    Allefs, J.J.H.M.

    1995-01-01

    Blackleg is a disease of potato, Solanum tuberosum , which is caused by the bacteria Erwinia carotovora subsp. carotovora ( Ecc ), E.c. subsp. atroseptica ( Eca ) or E . chrysanthemi (Ech). Incidence of blackleg negatively affects the quality of seed potatoes. Disease control relies on phytosanitary

  8. 不同产地野菊花中绿原酸、4,5-二咖啡酰奎尼酸的含量测定%Content Determination of Chlorogenic Acid and 4,5-O-Dicaffeoylquinic Acid in Chrysanthemi Indici Flos from Different Areas

    Institute of Scientific and Technical Information of China (English)

    崔永霞; 吴明侠; 许闽; 王丽果

    2012-01-01

    OBJECTIVE To determine the content of chlorogenic acid and 4,5-O-dicaffeoylquinic acid in Chrysanthemi Indici Flos from different areas. METHODS HPLC was used. The chromatographic separation was performed on a Diamonsil C18(4.6 mm × 250 mm,5 μm) column with gradient elute. The detection wavelength was 327 nm and column temperature was 30℃. RESULTS Good linear relationship between the peak area and content of chlorogenic acid and 4,5-O-dicaffeoylquinic acid was achieved when the amount was within 0.011-0.22 ug,0.0175-0.35 μg; the recoveries were 99.1% and 98.8%; RSD were 1.93% and 1.54%(n=9). CONCLUSION The method is accurate,reproducible and suitable to determine chlorogenic acid and 4,5-O-dicaffeoylquinic acid in Chrysanthemi Indici Flos.%目的 建立不同产地野菊花中绿原酸、4,5-二咖啡酰奎尼酸的含量测定方法.方法 采用HPLC,色谱柱为Diamonsil C18(4.6mm×250 mm,5μm);采用梯度洗脱;检测波长327 nm;柱温30℃;流速1.0 mL·min-1.结果 绿原酸、4,5-二咖啡酰奎尼酸的线性范围分别为0.011~0.22 μg,0.0175~0.35 μg.加样回收率分别为99.1%和98.8%; RSD分别为1.93%和1.54%(n=9).结论 本方法操作简便,分析速度快,结果准确,重复性好,可用于野菊花中绿原酸和4,5-二咖啡酰奎尼酸含量的测定.

  9. Identification of genes differentially expressed during interaction of resistant and susceptible apple cultivars (Malus × domestica with Erwinia amylovora

    Directory of Open Access Journals (Sweden)

    Aldwinckle Herb S

    2010-01-01

    Full Text Available Abstract Background The necrogenic enterobacterium, Erwinia amylovora is the causal agent of the fire blight (FB disease in many Rosaceaespecies, including apple and pear. During the infection process, the bacteria induce an oxidative stress response with kinetics similar to those induced in an incompatible bacteria-plant interaction. No resistance mechanism to E. amylovora in host plants has yet been characterized, recent work has identified some molecular events which occur in resistant and/or susceptible host interaction with E. amylovora: In order to understand the mechanisms that characterize responses to FB, differentially expressed genes were identified by cDNA-AFLP analysis in resistant and susceptible apple genotypes after inoculation with E. amylovora. Results cDNA were isolated from M.26 (susceptible and G.41 (resistant apple tissues collected 2 h and 48 h after challenge with a virulent E. amylovora strain or mock (buffer inoculated. To identify differentially expressed transcripts, electrophoretic banding patterns were obtained from cDNAs. In the AFLP experiments, M.26 and G.41 showed different patterns of expression, including genes specifically induced, not induced, or repressed by E. amylovora. In total, 190 ESTs differentially expressed between M.26 and G.41 were identified using 42 pairs of AFLP primers. cDNA-AFLP analysis of global EST expression in a resistant and a susceptible apple genotype identified different major classes of genes. EST sequencing data showed that genes linked to resistance, encoding proteins involved in recognition, signaling, defense and apoptosis, were modulated by E. amylovora in its host plant. The expression time course of some of these ESTs selected via a bioinformatic analysis has been characterized. Conclusion These data are being used to develop hypotheses of resistance or susceptibility mechanisms in Malus to E. amylovora and provide an initial categorization of genes possibly involved in

  10. Evaluation of kasugamycin for fire blight management, effect on nontarget bacteria, and assessment of kasugamycin resistance potential in Erwinia amylovora.

    Science.gov (United States)

    McGhee, Gayle C; Sundin, George W

    2011-02-01

    The emergence and spread of streptomycin-resistant strains of Erwinia amylovora in Michigan has necessitated the evaluation of new compounds effective for fire blight control. The aminoglycoside antibiotic kasugamycin (Ks) targets the bacterial ribosome and is particularly active against E. amylovora. The efficacy of Ks formulated as Kasumin 2L for control of fire blight was evaluated in six experiments conducted over four field seasons in our experimental orchards in East Lansing, MI. Blossom blight control was statistically equivalent to the industry standard streptomycin in all experiments. E. amylovora populations remained constant on apple flower stigmas pretreated with Kasumin and were ≈100-fold lower than on stigmas treated with water. Kasumin applied to apple trees in the field also resulted in a 100-fold reduced total culturable bacterial population compared with trees treated with water. We performed a prospective analysis of the potential for kasugamycin resistance (Ks(R)) development in E. amylovora which focused on spontaneous resistance development and acquisition of a transferrable Ks(R) gene. In replicated lab experiments, the development of spontaneous resistance in E. amylovora to Ks at 250 or 500 ppm was not observed when cells were directly plated on medium containing high concentrations of the antibiotic. However, exposure to increasing concentrations of Ks in media (initial concentration 25 μg ml(-1)) resulted in the selection of Ks resistance (at 150 μg ml(-1)) in the E. amylovora strains Ea110, Ea273, and Ea1189. Analysis of mutants indicated that they harbored mutations in the kasugamycin target ksgA gene and that all mutants were impacted in relative fitness observable through a reduced growth rate in vitro and decreased virulence in immature pear fruit. The possible occurrence of a reservoir of Ks(R) genes in orchard environments was also examined. Culturable gram-negative bacteria were surveyed from six experimental apple orchards

  11. 响应面法优化广西红树林菌种Erwinia sp.5-8产抗菌活性物质的发酵条件%Optimization of fermentation medium and conditions of antibiotic active substances production by the Erwinia sp.5-8 isolated from Guangxi mangrove

    Institute of Scientific and Technical Information of China (English)

    邱晨; 孟令洋; 张培玉

    2015-01-01

    目的 采用响应面法(RSM)优化广西红树林菌种Erwinia sp.5-8产抗菌活性物质的发酵条件.方法 在单因素实验最适发酵条件基础上,利用Plackett-Burman筛选出对抑菌活性有显著影响的3个因素为温度、盐度和蛋白胨的浓度浓度,在此基础上通过最陡爬坡实验逼近最佳响应面区域;再运用Box-Behnken实验设计和响应面分析法进行回归分析,确定重要因素的最优条件.结果 菌株Erwinia5-8的最佳发酵条件为:温度为26.0℃,盐度为0.96%,蛋白胨的浓度为0.27%,葡萄糖浓度为0.6%,初始pH为8.5,装液量为40%(V/V),接种量为1%.结论 在此最优条件下菌株Erwinia 5-8发酵液的抗菌活性较优化前提高了7.96%.

  12. Prokaryotic expression of iceA gene from ice nucleation active bacteria Erwinia ananas 110 and analysis of ice nucleation activity%冰核细菌Erwinia ananas 110冰核基因iceA的原核表达及冰核活性分析

    Institute of Scientific and Technical Information of China (English)

    姚润贤; 袁哲明

    2013-01-01

    To obtain recombinant strain with high ice nucleation activity,iceA gene were amplified by PCR from ice nucleation active bacteria Erwinia ananas 110 and cloned into vector pMD19-T which was transformed into E.coli DH5α.The recombinant clones were screened by single and double digestion before sequenced.From the positive recombinant strain,iceA gene was subcloned into prokaryotic expression vector pET-23a(+),resulting in recombinant plasmid pET-23a(+)-ice which was transformed into E.coli BL21(DE3)pLysS and induced by IPTG.SDS-PAGE indicated that ice nucleation active protein was expressed as inclusion bodies with molecular weight of about 180 000.Ice nucleation activity test showed there was no difference in ice nucleation activity under-5,4,-3,and-2 ℃ between recombinant E.coli BL21(DE3)pLysS and wild ice nucleation active bacteria Erwinia ananas 110.%为获得具有高冰核活性的基因工程菌,从冰核细菌Erwinia ananas 110扩增冰核基因iceA,将其克隆到pMD 19-T载体上,转化大肠杆菌DH5α,单、双酶切鉴定并测序;阳性克隆目的片段亚克隆到表达载体pET-23a(+)上,转化大肠杆菌DH5αt,单、双酶切鉴定重组质粒;阳性重组质粒转化大肠杆菌BL21(DE3)pLysS,并经IPTG诱导表达.SDS-PAGE电泳检测表明,冰核基因iceA能够并以包涵体形式表达,相对分子质量约为180 000.冰核活性测定结果表明,重组菌BL21 (DE3)pLysS/pET-ice的冰核活性与野生冰核细菌Erwinia ananas 110在-5、-4、-3、-2℃下无明显差别.

  13. Rock Phosphate Solubilization Mechanisms of One Fungus and One Bacterium

    Institute of Scientific and Technical Information of China (English)

    LIN Qi-mei; ZHAO Xiao-rong; ZHAO Zi-juan; LI Bao-guo

    2002-01-01

    Many microorganisms can dissolve the insoluble phosphates like apatite. However, the mechanisms are still not clear. This study was an attempt to investigate the mechanisms of rock phosphate solubilization by an Aspergillus 2TCiF2 and an Arthrobacter1TCRi7. The results indicated that the fungus produced a large amount of organic acids, mainly oxalic acid. The total quantity of the organic acids produced by the fungus was 550 times higher than that by the bacterium. Different organic acids had completely different capacities to solubilize the rock. Oxalic acid and citric acid had stronger capacity to dissolve the rock than malic acid, tartaric acid, lactic acid, acetic acid, malonic acid and succinic acid. The fungus solubilized the rock through excreting both proton and organic acids. The rock solubilization of the bacterium depended on only proton.

  14. A Streamlined Strategy for Biohydrogen Production with an Alkaliphilic Bacterium

    Energy Technology Data Exchange (ETDEWEB)

    Elias, Dwayne A [ORNL; Wall, Judy D. [University of Missouri; Mormile, Dr. Melanie R. [Missouri University of Science and Technology; Begemann, Matthew B [University of Wisconsin, Madison

    2012-01-01

    Biofuels are anticipated to enable a shift from fossil fuels for renewable transportation and manufacturing fuels, with biohydrogen considered attractive since it could offer the largest reduction of global carbon budgets. Currently, biohydrogen production remains inefficient and heavily fossil fuel-dependent. However, bacteria using alkali-treated biomass could streamline biofuel production while reducing costs and fossil fuel needs. An alkaliphilic bacterium, Halanaerobium strain sapolanicus, is described that is capable of biohydrogen production at levels rivaling neutrophilic strains, but at pH 11 and hypersaline conditions. H. sapolanicus ferments a variety of 5- and 6- carbon sugars derived from hemicellulose and cellulose including cellobiose, and forms the end products hydrogen and acetate. Further, it can also produce biohydrogen from switchgrass and straw pretreated at temperatures far lower than any previously reported and in solutions compatible with growth. Hence, this bacterium can potentially increase the efficiency and efficacy of biohydrogen production from renewable biomass resources.

  15. Initiation of chromosomal replication in predatory bacterium Bdellovibrio bacteriovorus

    Directory of Open Access Journals (Sweden)

    Lukasz Makowski

    2016-11-01

    Full Text Available Bdellovibrio bacteriovorus is a small Gram-negative predatory bacterium that attacks other Gram-negative bacteria, including many animal, human, and plant pathogens. This bacterium exhibits a peculiar biphasic life cycle during which two different types of cells are produced: non-replicating highly motile cells (the free-living phase and replicating cells (the intracellular-growth phase. The process of chromosomal replication in B. bacteriovorus must therefore be temporally and spatially regulated to ensure that it is coordinated with cell differentiation and cell cycle progression. Recently, B. bacteriovorus has received considerable research interest due to its intriguing life cycle and great potential as a prospective antimicrobial agent. Although we know that chromosomal replication in bacteria is mainly regulated at the initiation step, no data exists about this process in B. bacteriovorus. We report the first characterization of key elements of initiation of chromosomal replication – DnaA protein and oriC region from the predatory bacterium, B. bacteriovorus. In vitro studies using different approaches demonstrate that the B. bacteriovorus oriC (BdoriC is specifically bound and unwound by the DnaA protein. Sequence comparison of the DnaA-binding sites enabled us to propose a consensus sequence for the B. bacteriovorus DnaA box (5’-NN(A/TTCCACA-3’. Surprisingly, in vitro analysis revealed that BdoriC is also bound and unwound by the host DnaA proteins (relatively distantly related from B. bacteriovorus. We compared the architecture of the DnaA–oriC complexes (orisomes in homologous (oriC and DnaA from B. bacteriovorus and heterologous (BdoriC and DnaA from prey, E. coli or P. aeruginosa systems. This work provides important new entry points toward improving our understanding of the initiation of chromosomal replication in this predatory bacterium.

  16. Biosorption of heavy metals by a marine bacterium

    Energy Technology Data Exchange (ETDEWEB)

    Iyer, Anita [Central Salt and Marine Chemicals Research Institute, Bhavnagar 364002, Gujarat (India); Mody, Kalpana [Central Salt and Marine Chemicals Research Institute, Bhavnagar 364002, Gujarat (India)]. E-mail: khmody@csmcri.org; Jha, Bhavanath [Central Salt and Marine Chemicals Research Institute, Bhavnagar 364002, Gujarat (India)

    2005-03-01

    Heavy metal chelation property of exopolysaccharide produced by Enterobacter cloaceae, a marine bacterium, isolated from the West Coast of India, is reported in this paper. The exopolysaccharide demonstrated excellent chelating properties with respect to cadmium (65%) followed by copper (20%) and cobalt (8%) at 100 mg/l heavy metal concentration. However, it could not chelate mercury. A comparative study of the percentage biosorption of the above mentioned metals is presented here.

  17. Growth of a Strictly Anaerobic Bacterium on Furfural (2-Furaldehyde)

    OpenAIRE

    Brune, Gerhard; Schoberth, Siegfried M.; Sahm, Hermann

    1983-01-01

    A strictly anaerobic bacterium was isolated from a continuous fermentor culture which converted the organic constituents of sulfite evaporator condensate to methane and carbon dioxide. Furfural is one of the major components of this condensate. This furfural isolate could degrade furfural as the sole source of carbon and energy in a defined mineral-vitamin-sulfate medium. Acetic acid was the major fermentation product. This organism could also use ethanol, lactate, pyruvate, or fumarate and c...

  18. HrpG and HrpV proteins from the Type III secretion system of Erwinia amylovora form a stable heterodimer.

    Science.gov (United States)

    Gazi, Anastasia D; Charova, Spyridoula; Aivaliotis, Michalis; Panopoulos, Nicholas J; Kokkinidis, Michael

    2015-01-01

    Bacterial type III secretion systems (T3SSs) are specialized multicomponent nanomachines that mediate the transport of proteins either to extracellular locations or directly into eukaryotic host cell cytoplasm. Erwinia amylovora, the main agent of rosaceous plants fireblight disease, employs an Hrp/Hrc1 T3SS to accomplish its pathogenesis. The regulatory network that controls the activation of this T3SS is largely unknown in E. amylovora. However, in Pseudomonas syringae pathovars, the HrpG/HrpV complex has been shown to directly regulate the activity of transcription factor HrpS and consequently the upregulation of the Hrp/Hrc1 T3SS related genes. In this work, we report the successful recombinant production and purification of a stable E. amylovora HrpG/HrpV complex, using pPROpET, a bicistronic expression vector. Furthermore, we present the first solution structure of this complex based on small-angle X-ray scattering data.

  19. Halogenated furanones from the red alga, Delisea pulchra, inhibit carbapenem antibiotic synthesis and exoenzyme virulence factor production in the phytopathogen Erwinia carotovora

    DEFF Research Database (Denmark)

    Manefield, M.; Welch, M.; Givskov, Michael Christian

    2001-01-01

    The plant pathogen Erwinia carotovora regulates expression of virulence factors and antibiotic production via an N-3- oxohexanoyl-L-homoserine lactone (3-oxo-C6-HSL) dependent quorum sensing mechanism. The marine alga Delisea pulchra produces halogenated furanones known to antagonise 3-oxo-C6-HSL...... activity. We have tested the effects of a halogenated furanone on the production of carbapenem, cellulase and protease in E. carotovora. Despite differences in the regulatory mechanisms controlling carbapenem and exoenzyme production each was inhibited by the algal metabolite. We present evidence...... to suggest that the furanone dependent inhibition of carbapenem production is a result of the disruption of the 3-oxo-C6-HSL dependent expression of the carABCDEFGH operon....

  20. 甘薯细菌性黑腐病发生流行的研究%Occurrence and epidemic of Erwinia carotovora sp in sweet potato

    Institute of Scientific and Technical Information of China (English)

    罗克昌; 李云平; 陈路招; 林意; 陈聚元

    2003-01-01

    对甘薯细菌性黑腐病(Erwinia carotovora sp)的发生流行研究结果表明:土壤中残存的病株是病害的初侵染源,病菌从耕作栽培伤口侵入,土壤过湿和高温多雨气候是病害流行条件.甘薯栽种后至茎叶旺长结薯期的50~60天是病害对鲜薯产量影响的关键时期,也是防治的重要时期,病株率与产量成显著负相关(相关系数为-0.8828).

  1. [The feasibility of Erwinia asparaginase for pediatric patients who developed an allergic reaction to E.coli asparaginase during treatment of acute lymphoblastic leukemia].

    Science.gov (United States)

    Takahashi, Hiroyoshi; Koh, Katsuyoshi; Kato, Motohiro; Isobe, Kiyotaka; Yasui, Naoko; Mori, Makiko; Akiyama, Kosuke; Kikuchi, Akira; Hanada, Ryoji

    2013-04-01

    Asparaginase (ASNase) is one of the most important key drugs in the treatment of acute lymphoblastic leukemia (ALL). However, clinical hypersensitivity reactions often occur and lead to the discontinuation of ASNase treatment. Here, we report a retrospective study of 68 Erwinia ASNase (Erw-ASNase) administrations in 11 patients with childhood ALL who developed allergic reactions to E.coli-ASNase in our hospital between 2006 and 2012. The median age of the patients was 6 (range, 0 to 14). Erw-ASNase purchased overseas by the patients' guardians had already been administered when we obtained informed consent from the guardians. In all patients, fibrinogen and/or anti-thrombin III levels were decreased, but thrombosis did not develop. There was only one mild adverse event (grade 2 urticaria) in one patient, in whom Erw-ASNase could be continued after increasing the doses of premedication with antihistamine and prednisolone. Erw-ASNase could be safely administered to all patients.

  2. Systems level analysis of two-component signal transduction systems in Erwinia amylovora: Role in virulence, regulation of amylovoran biosynthesis and swarming motility

    Directory of Open Access Journals (Sweden)

    Sundin George W

    2009-05-01

    Full Text Available Abstract Background Two-component signal transduction systems (TCSTs, consisting of a histidine kinase (HK and a response regulator (RR, represent a major paradigm for signal transduction in prokaryotes. TCSTs play critical roles in sensing and responding to environmental conditions, and in bacterial pathogenesis. Most TCSTs in Erwinia amylovora have either not been identified or have not yet been studied. Results We used a systems approach to identify TCST and related signal transduction genes in the genome of E. amylovora. Comparative genomic analysis of TCSTs indicated that E. amylovora TCSTs were closely related to those of Erwinia tasmaniensis, a saprophytic enterobacterium isolated from apple flowers, and to other enterobacteria. Forty-six TCST genes in E. amylovora including 17 sensor kinases, three hybrid kinases, 20 DNA- or ligand-binding RRs, four RRs with enzymatic output domain (EAL-GGDEF proteins, and two kinases were characterized in this study. A systematic TCST gene-knockout experiment was conducted, generating a total of 59 single-, double-, and triple-mutants. Virulence assays revealed that five of these mutants were non-pathogenic on immature pear fruits. Results from phenotypic characterization and gene expression experiments indicated that several groups of TCST systems in E. amylovora control amylovoran biosynthesis, one of two major virulence factors in E. amylovora. Both negative and positive regulators of amylovoran biosynthesis were identified, indicating a complex network may control this important feature of pathogenesis. Positive (non-motile, EnvZ/OmpR, negative (hypermotile, GrrS/GrrA, and intermediate regulators for swarming motility in E. amylovora were also identified. Conclusion Our results demonstrated that TCSTs in E. amylovora played major roles in virulence on immature pear fruit and in regulating amylovoran biosynthesis and swarming motility. This suggested presence of regulatory networks governing

  3. Characterization of the rcsB gene from Erwinia amylovora and its influence on exoploysaccharide synthesis and virulence of the fire blight pathogen.

    Science.gov (United States)

    Bereswill, S; Geider, K

    1997-02-01

    RcsB belongs to a family of positive regulators of exopolysaccharide synthesis in various enterobacteria. The rcsB gene of the fire blight pathogen Erwinia amylovora was cloned by PCR amplification with consensus primers, and its role in exopolysaccharide (EPS) synthesis was investigated. Its overexpression from high-copy-number plasmids stimulated the synthesis of the acidic EPS amylovoran and suppressed expression of the levan-forming enzyme levansucrase. Inactivation of rcsB by site-directed mutagenesis created mutants that were deficient in amylovoran synthesis and avirulent on host plants. In addition, a cosmid which complemented rcsB mutants was selected from a genomic library. The spontaneous E. amylovora mutant E8 has a similar phenotype and was complemented by the cloned rcsB gene. The rcsB region of strain E8 was also amplified by PCR, and the mutation was characterized as a nine-nucleotide deletion at the start of the rcsB gene. Nucleotide sequence analysis of the E. amylovora rcsB region and the predicted amino acid sequence of RcsB revealed extensive homology to rcsB and the encoded protein of other bacteria such as Escherichia coli and Erwinia stewartii. In all three organisms, rcsB is localized adjacent to the rcsC gene, which is transcribed in the opposite direction of rcsB. The E. amylovora rcsB gene has now been shown to strongly affect the formation of disease symptoms of a plant pathogen.

  4. Erwinia carotovora DsbA mutants: evidence for a periplasmic-stress signal transduction system affecting transcription of genes encoding secreted proteins.

    Science.gov (United States)

    Vincent-Sealy, L V; Thomas, J D; Commander, P; Salmond, G P

    1999-08-01

    The dsbA genes, which encode major periplasmic disulfide-bond-forming proteins, were isolated from Erwinia carotovora subsp. carotovora (Ecc) and Erwinia carotovora subsp. atroseptica (Eca), and the dsbC gene, encoding another periplasmic disulfide oxidoreductase was isolated from Ecc. All three genes were sequenced and mutants deficient in these genes were created by marker exchange mutagenesis. The Ecc mutants were severely affected in activity and secretion of pectate lyase, probably due to the absence of functional PelC, which is predicted to require disulfide bond formation to achieve its correct conformation prior to secretion across the outer membrane. Similarly, endopolygalacturonase, also predicted to possess disulfide bonds, displayed reduced activity. The major Ecc cellulase (CelV) does not contain cysteine residues and was still secreted in dsbA-deficient strains. This observation demonstrated unequivocally that the localization and activity of the individual components of the Out apparatus are independent of disulfide bond formation. Surprisingly, cellulase activity was shown to be increased approximately two- to threefold in the DsbA mutant. This phenomenon resulted from transcriptional up-regulation of celV gene expression. In contrast, transcription of both pelC and peh were down-regulated in dsbA-deficient strains when compared to the wild-type. Protease (Prt) activity and secretion were unaffected in the Ecc dsbA mutant. Prt activity was considerably reduced in the double dsbA dsbC mutant. However Prt was secreted normally in this strain. The Eca dsbA mutant was found to be non-motile, suggesting that disulfide bond formation is essential for motility in this strain. All of the dsb mutants showed reduced tissue maceration in planta. These results suggest that a feedback regulation system operates in Ecc. In this system, defects in periplasmic disulfide bond formation act as a signal which is relayed to the transcription machinery regulating gene

  5. Mutagenesis and functional characterization of the RNA and protein components of the toxIN abortive infection and toxin-antitoxin locus of Erwinia.

    Science.gov (United States)

    Blower, T R; Fineran, P C; Johnson, M J; Toth, I K; Humphreys, D P; Salmond, G P C

    2009-10-01

    Bacteria are constantly challenged by bacteriophage (phage) infection and have developed multiple adaptive resistance mechanisms. These mechanisms include the abortive infection systems, which promote "altruistic suicide" of an infected cell, protecting the clonal population. A cryptic plasmid of Erwinia carotovora subsp. atroseptica, pECA1039, has been shown to encode an abortive infection system. This highly effective system is active across multiple genera of gram-negative bacteria and against a spectrum of phages. Designated ToxIN, this two-component abortive infection system acts as a toxin-antitoxin module. ToxIN is the first member of a new type III class of protein-RNA toxin-antitoxin modules, of which there are multiple homologues cross-genera. We characterized in more detail the abortive infection phenotype of ToxIN using a suite of Erwinia phages and performed mutagenesis of the ToxI and ToxN components. We determined the minimal ToxI RNA sequence in the native operon that is both necessary and sufficient for abortive infection and to counteract the toxicity of ToxN. Furthermore, site-directed mutagenesis of ToxN revealed key conserved amino acids in this defining member of the new group of toxic proteins. The mechanism of phage activation of the ToxIN system was investigated and was shown to have no effect on the levels of the ToxN protein. Finally, evidence of negative autoregulation of the toxIN operon, a common feature of toxin-antitoxin systems, is presented. This work on the components of the ToxIN system suggests that there is very tight toxin regulation prior to suicide activation by incoming phage.

  6. Biotransformation of citrinin to decarboxycitrinin using an organic solvent-tolerant marine bacterium, Moraxella sp. (MB1)

    Digital Repository Service at National Institute of Oceanography (India)

    PrabhaDevi; Naik, C.G.; Rodrigues, C.

    . In the present study, we used an organic solvent-tolerant marine bacterium, Moraxella sp. MB1. 16S rRNA sequencing revealed that the bacterium shows 98% similarity with an uncultured marine bacterium with gene bank accession number AY936933. This bacterium...

  7. Salt-inducible promoter derivable from a lactic acid bacterium, and its use in a lactic acid bacterium for production of a desired protein

    NARCIS (Netherlands)

    Sanders, Jan Willem; Kok, Jan; Venema, Gerard; Ledeboer, Adrianus Marinus

    1998-01-01

    The invention provides a salt-inducible promoter present in SEQ ID NO: 10 and derivable from a lactic acid bacterium in isolation from the coding sequence normally controlled by said promoter in a wild-type lactic acid bacterium, with modifications and important parts thereof. Also provided are a re

  8. Purification of Ice-Nucleation Active Proteins of Ice- Nucleation Active Bacteria Erwinia herbicola%冰核活性细菌中冰核活性蛋白质的分离与纯化的研究

    Institute of Scientific and Technical Information of China (English)

    陈庆森; 王昌禄; 高秀芝; 阎亚丽; 庞广昌

    2007-01-01

    解决生物冰核应用中的安全问题,从产冰核活性的细菌中分离纯化天然冰核活性蛋白质是一条重要途径,对其应用具有重要价值.本研究以冰核活性细菌(Erwinia herbicola 10025A)为实验菌种,经培养获得高浓度发酵液,离心处理后,采用超声波破碎、凝胶层析的方法,分离纯化得到了Erwinia herbicola 10025A的Ⅰ型冰核活性蛋白质.该蛋白质经SDS-PAGE的分析鉴定,证实了该冰核活性蛋白质的分子量为150kD左右.

  9. Interaction entre des mutants hrp d'Erwinia amylovora, agent du feu bactérien, le parent pathogène et la plante hôte : recherche de mécanismes modulant la compatibilité

    OpenAIRE

    Cesbron, Sophie

    2009-01-01

    Erwinia amylovora is the causal agent of fire blight, a disease that affects Maloideae. This gammaproteobacteria requires a type three secretion system (T3SS) for its pathogenicity. Previous work has shown that avirulent hrp mutant strains of E. amylovora affected in regulatory functions (hrpL and hrpS) protect apple seedlings from developping fire blight symptoms. We investigated molecular mechanisms leading to this protection. In a first part of our work, we studied molecular responses of t...

  10. Comparative genomics of the type VI secretion systems of Pantoea and Erwinia species reveals the presence of putative effector islands that may be translocated by the VgrG and Hcp proteins

    Directory of Open Access Journals (Sweden)

    De Maayer Pieter

    2011-11-01

    Full Text Available Abstract Background The Type VI secretion apparatus is assembled by a conserved set of proteins encoded within a distinct locus. The putative effector proteins Hcp and VgrG are also encoded within these loci. We have identified numerous distinct Type VI secretion system (T6SS loci in the genomes of several ecologically diverse Pantoea and Erwinia species and detected the presence of putative effector islands associated with the hcp and vgrG genes. Results Between two and four T6SS loci occur among the Pantoea and Erwinia species. While two of the loci (T6SS-1 and T6SS-2 are well conserved among the various strains, the third (T6SS-3 locus is not universally distributed. Additional orthologous loci are present in Pantoea sp. aB-valens and Erwinia billingiae Eb661. Comparative analysis of the T6SS-1 and T6SS-3 loci showed non-conserved islands associated with the vgrG and hcp, and vgrG genes, respectively. These regions had a G+C content far lower than the conserved portions of the loci. Many of the proteins encoded within the hcp and vgrG islands carry conserved domains, which suggests they may serve as effector proteins for the T6SS. A number of the proteins also show homology to the C-terminal extensions of evolved VgrG proteins. Conclusions Extensive diversity was observed in the number and content of the T6SS loci among the Pantoea and Erwinia species. Genomic islands could be observed within some of T6SS loci, which are associated with the hcp and vgrG proteins and carry putative effector domain proteins. We propose new hypotheses concerning a role for these islands in the acquisition of T6SS effectors and the development of novel evolved VgrG and Hcp proteins.

  11. Influence of plaque-forming bacterium, Rhodobacteraceae sp. on the growth of Chlorella vulgaris.

    Science.gov (United States)

    Chen, Zhangran; Zhang, Jingyan; Lei, Xueqian; Zhang, Bangzhou; Cai, Guanjing; Zhang, Huajun; Li, Yi; Zheng, Wei; Tian, Yun; Xu, Hong; Zheng, Tianling

    2014-10-01

    Experiments were conducted to find out the molecular features, infection process of a special alga plaque-forming microorganism and its potential influence on the biomass of Chlorella vulgaris during the infection process. Direct contact between the algal cell and the bacterium may be the primary steps needed for the bacterium to lyse the alga. Addition of C. vulgaris cells into f/2 medium allowed us obtain the object bacterium. The 16S rRNA gene sequence comparisons results showed that the plaque-forming bacterium kept the closest relationship with Labrenzia aggregata IAM 12614(T) at 98.90%. The existence of the bacterium could influence both the dry weight and lipid content of C. vulgaris. This study demonstrated that direct cell wall disruption of C. vulgaris by the bacterium would be a potentially effective method to utilize the biomass of microalgae.

  12. Research Progress and Perspectives of Nitrogen Fixing Bacterium, Gluconacetobacter diazotrophicus, in Monocot Plants

    Directory of Open Access Journals (Sweden)

    N. Eskin

    2014-01-01

    Full Text Available Gluconacetobacter diazotrophicus is a nitrogen fixing bacterium originally found in monocotyledon sugarcane plants in which the bacterium actively fixes atmosphere nitrogen and provides significant amounts of nitrogen to plants. This bacterium mainly colonizes intercellular spaces within the roots and stems of plants and does not require the formation of the complex root organ like nodule. The bacterium is less plant/crop specific and indeed G. diazotrophicus has been found in a number of unrelated plant species. Importantly, as the bacterium was of monocot plant origin, there exists a possibility that the nitrogen fixation feature of the bacterium may be used in many other monocot crops. This paper reviews and updates the research progress of G. diazotrophicus for the past 25 years but focuses on the recent research development.

  13. Factors Affecting Zebra Mussel Kill by the Bacterium Pseudomonas fluorescens

    Energy Technology Data Exchange (ETDEWEB)

    Daniel P. Molloy

    2004-02-24

    The specific purpose of this research project was to identify factors that affect zebra mussel kill by the bacterium Pseudomonas fluorescens. Test results obtained during this three-year project identified the following key variables as affecting mussel kill: treatment concentration, treatment duration, mussel siphoning activity, dissolved oxygen concentration, water temperature, and naturally suspended particle load. Using this latter information, the project culminated in a series of pipe tests which achieved high mussel kill inside power plants under once-through conditions using service water in artificial pipes.

  14. Liver abscess associated with an oral flora bacterium Streptococcus anginosus

    Directory of Open Access Journals (Sweden)

    Hava Yılmaz

    2012-03-01

    Full Text Available Viridans group Streptococcus, a bacterium of the oral flora has a low-virulence and rarely causes liver abscess. A 40-yearoldmale patient was admitted to the hospital complaining of high fever and malaise. A physical examination revealedpoor oral hygiene; there were caries on many teeth, and he had hepatomegaly. A hepatic abscess was identified inhis abdominal tomography. Streptococcus anginosus was isolated from the drainage material, and the bile ducts werenormal in his MRI cholangiography. An immunocompetent case of liver abscess caused by Streptococcus anginosusoriginated most probably from oral flora is presented here. J Microbiol Infect Dis 2012; 2(1:33-35

  15. Biological Control of Meloidogyne hapla Using an Antagonistic Bacterium

    Directory of Open Access Journals (Sweden)

    Jiyeong Park

    2014-09-01

    Full Text Available We examined the efficacy of a bacterium for biocontrol of the root-knot nematode (RKN Meloidogyne hapla in carrot (Daucus carota subsp. sativus and tomato (Solanum lycopersicum. Among 542 bacterial isolates from various soils and plants, the highest nematode mortality was observed for treatments with isolate C1-7, which was identified as Bacillus cereus based on cultural and morphological characteristics, the Biolog program, and 16S rRNA sequencing analyses. The population density and the nematicidal activity of B. cereus C1-7 remained high until the end of culture in brain heart infusion broth, suggesting that it may have sustainable biocontrol potential. In pot experiments, the biocontrol efficacy of B. cereus C1-7 was high, showing complete inhibition of root gall or egg mass formation by RKN in carrot and tomato plants, and subsequently reducing RKN damage and suppressing nematode population growth, respectively. Light microscopy of RKN-infected carrot root tissues treated with C1-7 showed reduced formation of gall cells and fully developed giant cells, while extensive gall cells and fully mature giant cells with prominent cell wall ingrowths formed in the untreated control plants infected with RKNs. These histopathological characteristics may be the result of residual or systemic biocontrol activity of the bacterium, which may coincide with the biocontrol efficacies of nematodes in pots. These results suggest that B. cereus C1-7 can be used as a biocontrol agent for M. hapla.

  16. Molybdate Reduction to Molybdenum Blue by an Antarctic Bacterium

    Directory of Open Access Journals (Sweden)

    S. A. Ahmad

    2013-01-01

    Full Text Available A molybdenum-reducing bacterium from Antarctica has been isolated. The bacterium converts sodium molybdate or Mo6+ to molybdenum blue (Mo-blue. Electron donors such as glucose, sucrose, fructose, and lactose supported molybdate reduction. Ammonium sulphate was the best nitrogen source for molybdate reduction. Optimal conditions for molybdate reduction were between 30 and 50 mM molybdate, between 15 and 20°C, and initial pH between 6.5 and 7.5. The Mo-blue produced had a unique absorption spectrum with a peak maximum at 865 nm and a shoulder at 710 nm. Respiratory inhibitors such as antimycin A, sodium azide, potassium cyanide, and rotenone failed to inhibit the reducing activity. The Mo-reducing enzyme was partially purified using ion exchange and gel filtration chromatography. The partially purified enzyme showed optimal pH and temperature for activity at 6.0 and 20°C, respectively. Metal ions such as cadmium, chromium, copper, silver, lead, and mercury caused more than 95% inhibition of the molybdenum-reducing activity at 0.1 mM. The isolate was tentatively identified as Pseudomonas sp. strain DRY1 based on partial 16s rDNA molecular phylogenetic assessment and the Biolog microbial identification system. The characteristics of this strain would make it very useful in bioremediation works in the polar and temperate countries.

  17. Molybdate reduction to molybdenum blue by an Antarctic bacterium.

    Science.gov (United States)

    Ahmad, S A; Shukor, M Y; Shamaan, N A; Mac Cormack, W P; Syed, M A

    2013-01-01

    A molybdenum-reducing bacterium from Antarctica has been isolated. The bacterium converts sodium molybdate or Mo⁶⁺ to molybdenum blue (Mo-blue). Electron donors such as glucose, sucrose, fructose, and lactose supported molybdate reduction. Ammonium sulphate was the best nitrogen source for molybdate reduction. Optimal conditions for molybdate reduction were between 30 and 50 mM molybdate, between 15 and 20°C, and initial pH between 6.5 and 7.5. The Mo-blue produced had a unique absorption spectrum with a peak maximum at 865 nm and a shoulder at 710 nm. Respiratory inhibitors such as antimycin A, sodium azide, potassium cyanide, and rotenone failed to inhibit the reducing activity. The Mo-reducing enzyme was partially purified using ion exchange and gel filtration chromatography. The partially purified enzyme showed optimal pH and temperature for activity at 6.0 and 20°C, respectively. Metal ions such as cadmium, chromium, copper, silver, lead, and mercury caused more than 95% inhibition of the molybdenum-reducing activity at 0.1 mM. The isolate was tentatively identified as Pseudomonas sp. strain DRY1 based on partial 16s rDNA molecular phylogenetic assessment and the Biolog microbial identification system. The characteristics of this strain would make it very useful in bioremediation works in the polar and temperate countries.

  18. Pathogenesis of helicobacter pylori infection: Bacterium and host relationship

    Directory of Open Access Journals (Sweden)

    Sokić-Milutinović Aleksandra

    2004-01-01

    Full Text Available Helicobacter pylori (H. pylori colonizes the gastric mucosa of a half of the mankind. Duodenal ulcer is found in 15-25%, t gastric ulcer in 13%, while gastric adenocarcinoma develops in 1% of all infected individuals. Pathogenesis of H. pylori infection is related to the virulence factors of the bacterium, environmental (dietary habits, hygiene, stress and host factors (age, sex, blood type. Colonization of the gastric mucosa is related to the motility of the bacterium, presence of lipopolysacharide (LPS and various bacterial enzymes. Gastric mucosal injury is the result of H. pylori LPS, vacuolization cytotoxin (vacA, cytotoxin associated protein (cagA, heat shock proteins and factors responsible for neutrophil chemotaxis and activity. H. pylori colonizes the gastric mucosa and zones of ectopic gastric epithelium. H. pylori infection is transmitted via oral-oral, fecal-oral and iatrogenic way (during endoscopy. Higher prevalence of the infection is associated with lower socioeconomic level, lack of drinking water, and living in a community. Acute H. pylori gastritis is superficial pangastritis progressing into the chronic phase after 7-10 days. Gastric mucosal atrophy and intestinal metaplasia can develop during the course of H. pylori infection. Clearly defined factors that influence the outcome of H. pylori infection include bacterial strain, distribution of gastritis, acid secretion and gastric mucosal atrophy.

  19. Structure and morphology of magnetite anaerobically-produced by a marine magnetotactic bacterium and a dissimilatory iron-reducing bacterium

    Science.gov (United States)

    Sparks, N. H. C.; Mann, S.; Bazylinski, D. A.; Lovley, D. R.; Jannasch, H. W.; Frankel, R. B.

    1990-04-01

    Intracellular crystals of magnetite synthesized by cells of the magnetotactic vibroid organism, MV-1, and extracellular crystals of magnetite produced by the non-magnetotactic dissimilatory iron-reducing bacterium strain GS-15, were examined using high-resolution transmission electron microscopy, electron diffraction and 57Fe Mo¨ssbauer spectroscopy. The magnetotactic bacterium contained a single chain of approximately 10 crystals aligned along the long axis of the cell. The crystals were essentially pure stoichiometric magnetite. When viewed along the crystal long axis the particles had a hexagonal cross-section whereas side-on they appeared as rectangules or truncated rectangles of average dimension, 53 × 35 nm. These findings are explained in terms of a three-dimensional morphology comprising a hexagonal prism of 110 faces which are capped and truncated by 111 end faces. Electron diffraction and lattice imaging studies indicated that the particles were structurally well-defined single crystals. In contrast, magnetite particles produced by the strain, GS-15 were irregular in shape and had smaller mean dimensions (14 nm). Single crystals were imaged but these were not of high structural perfection. These results highlight the influence of intracellular control on the crystallochemical specificity of bacterial magnetites. The characterization of these crystals is important in aiding the identification of biogenic magnetic materials in paleomagnetism and in studies of sediment magnetization.

  20. ANTIBACTERIAL ACTIVITY OF THREE MEDICINAL PLANTS OF KUMAUN HIMALAYA AGAINST SOME PATHOGENIC BACTERIA

    Directory of Open Access Journals (Sweden)

    S. C. SATI

    2015-11-01

    Full Text Available The antibacterial property of methanol, ethanol and hexane extracts of Berberis aristata, Chenopodium ambrosioides and Tinospora cordifolia grown in Kumaun Himalayan were investigated against some pathogenic gram positive and gram negative bacterial strains (Bacillus subtilis, Agrobacterium tumefaciens, Escherichia coli, Xanthomonas phaseoli and Erwinia chrysanthemi using disc diffusion method. Methanol extract of B. aristata was found with highest inhibitory activity against E. chrysanthemi (ZOI, 11±0.3mm. Whereas lowest inhibition was recorded in ethanolic extract of B. aristata against E. coli. The hexane extract of B. aristata and methanolic extract of C. ambrosioides were found totally inactive against all the pathogens tested.

  1. Dense populations of a giant sulfur bacterium in Namibian shelf sediments

    DEFF Research Database (Denmark)

    Schulz, HN; Brinkhoff, T.; Ferdelman, TG

    1999-01-01

    A previously unknown giant sulfur bacterium is abundant in sediments underlying the oxygen minimum zone of the Benguela Current upwelling system. The bacterium has a spherical cell that exceeds by up to 100-fold the biovolume of the largest known prokaryotes. On the basis of 16S ribosomal DNA seq...

  2. Hydrogen production by co-cultures of Lactobacillus and a photosynthetic bacterium, Rhodobacter sphaeroides RV

    Energy Technology Data Exchange (ETDEWEB)

    Asada, Yasuo; Ishimi, Katsuhiro [Department of General Education, College of Science and Technology, Nihon University, Narashinodai, Chiba 274-8501 (Japan); Tokumoto, Masaru; Aihara, Yasuyuki; Oku, Masayo; Kohno, Hideki [Department of Applied Molecular Chemistry, College of Industrial Technology, Nihon University, Izumi-cho, Chiba 275-8575 (Japan); Wakayama, Tatsuki; Miyake, Jun [Research Institute for Cell Engineering, National Institute of Advanced Industrial Science and Technology, Nakoji, Amagasaki, Hyogo 661-0974 (Japan); Tomiyama, Masamitsu [Genetic Diversity Department, National Institute of Agrobiological Science, Tsukuba, Ibaraki 305-8602 (Japan)

    2006-09-15

    Hydrogen production with glucose by using co-immobilized cultures of a lactic acid bacterium, Lactobacillus delbrueckii NBRC13953, and a photosynthetic bacterium, Rhodobacter sphaeroides RV, in agar gels was studied. Glucose was converted to hydrogen gas in a yield of 7.1mol of hydrogen per mole of glucose at a maximum under illuminated conditions. (author)

  3. Roseomonas gilardii subsp rosea, a pink bacterium associated with bacteremia: the first case in Thailand.

    Science.gov (United States)

    Srifuengfung, Somporn; Tharavichitkul, Prasit; Pumprueg, Satchana; Tribuddharat, Chanwit

    2007-09-01

    Roseomonas is a pink-pigmented, non-fermentative, gram-negative coccobacillus bacterium. Human infections caused by Roseomonas are very rare. We report the first case of bacteremia associated with Roseomonas gilardii subsp rosea in Thailand. The bacterium was isolated from blood culture and identified by cellular morphology, characteristics of colonies on blood agar, extensive biochemical tests and 16S ribosomal DNA sequencing.

  4. Genome Sequence of the Mycorrhizal Helper Bacterium Pseudomonas fluorescens BBc6R8

    OpenAIRE

    2014-01-01

    We report the draft genome sequence of the mycorrhizal helper bacterium Pseudomonas fluorescens strain BBc6R8. This is the first genome of a mycorrhizal helper bacterium. The draft genome contains 6,952,353 bp and is predicted to encode 6,317 open reading frames. Comparative genomic analyses will help to identify helper traits.

  5. Genome Sequence of the Mycorrhizal Helper Bacterium Pseudomonas fluorescens BBc6R8.

    Science.gov (United States)

    Deveau, A; Gross, H; Morin, E; Karpinets, T; Utturkar, S; Mehnaz, S; Martin, F; Frey-Klett, P; Labbé, J

    2014-01-09

    We report the draft genome sequence of the mycorrhizal helper bacterium Pseudomonas fluorescens strain BBc6R8. This is the first genome of a mycorrhizal helper bacterium. The draft genome contains 6,952,353 bp and is predicted to encode 6,317 open reading frames. Comparative genomic analyses will help to identify helper traits.

  6. Produção de isomaltulose a partir da transformação enzimática da sacarose, utilizando-se Erwinia sp D12 imobilizada com alginato de cálcio Production of isomaltulose from enzymatic transformation of sucrose, using Erwinia sp D12 immobilized with calcium alginate

    Directory of Open Access Journals (Sweden)

    Ana Lúcia Leite Moraes

    2005-03-01

    Full Text Available A glicosiltransferase de Erwinia sp D12 é capaz de converter a sacarose em isomaltulose (6-o-alfa-glicopiranosil D-frutofuranose, um açúcar alternativo que apresenta baixo potencial cariogênico, e que pode ser utilizado em chocolates, gomas de mascar e balas. A isomaltulose é também utilizada na produção de isomalte, uma mistura de açúcar álcool, de baixo valor calórico e baixo potencial cariogênico. No estudo da influência dos componentes do meio de cultivo, na produção de glicosiltransferase, em frascos agitados, foi obtido maior atividade da enzima (12,8 unidades de atividade/mL do meio de cultura em meio de cultura A constituído de melaço 12% (p/v de sólidos solúveis totais, peptona 4% (p/v e extrato de carne 0,4% (p/v. No estudo do efeito do tempo e da temperatura na fermentação da linhagem de Erwinia sp D12, em fermentador New Brunswick de 3L, contendo meio de cultura A, foi obtida maior atividade de glicosiltransferase (15,6 unidades de atividade/ mL de meio de cultura na fase exponencial de crescimento após 8 horas de fermentação a 30ºC. Na produção de isomaltulose a partir da sacarose utilizando-se células de Erwinia sp D12 imobilizadas em alginato de cálcio, estudou-se o efeito da temperatura (25 - 35ºC e da concentração de substrato (12,5 - 60% p/v. Foi obtido um rendimento em torno de 50% de isomaltulose, com soluções de sacarose entre 20-30% (p/v a 35ºC. Concentrações em excesso de sacarose (ao redor de 40% p/v afetaram a atividade da célula imobilizada, diminuindo a conversão de sacarose em isomaltulose. O xarope de isomaltulose foi purificado através de cromatografia de troca iônica e o eluato cristalizado por abaixamento de temperatura. Os cristais apresentaram 91,39% de isomaltulose.The glucosyltransferase of Erwinia sp D12 is able to convert sucrose into isomaltulose (6-0-alpha-D-glucopyranosyl-D-fructofuranose, an alternative sugar which presents low cariogenic potential, and can be

  7. The domestication of the probiotic bacterium Lactobacillus acidophilus.

    Science.gov (United States)

    Bull, Matthew J; Jolley, Keith A; Bray, James E; Aerts, Maarten; Vandamme, Peter; Maiden, Martin C J; Marchesi, Julian R; Mahenthiralingam, Eshwar

    2014-11-26

    Lactobacillus acidophilus is a Gram-positive lactic acid bacterium that has had widespread historical use in the dairy industry and more recently as a probiotic. Although L. acidophilus has been designated as safe for human consumption, increasing commercial regulation and clinical demands for probiotic validation has resulted in a need to understand its genetic diversity. By drawing on large, well-characterised collections of lactic acid bacteria, we examined L. acidophilus isolates spanning 92 years and including multiple strains in current commercial use. Analysis of the whole genome sequence data set (34 isolate genomes) demonstrated L. acidophilus was a low diversity, monophyletic species with commercial isolates essentially identical at the sequence level. Our results indicate that commercial use has domesticated L. acidophilus with genetically stable, invariant strains being consumed globally by the human population.

  8. Characterisation of an unusual bacterium isolated from genital ulcers.

    Science.gov (United States)

    Ursi, J P; van Dyck, E; Ballard, R C; Jacob, W; Piot, P; Meheus, A Z

    1982-02-01

    The preliminary characterisation of an unusual gram-negative bacillus isolated from genital ulcers in Swaziland is reported. Like Haemophilus ducreyi, it is an oxidase positive, nitrate-reductase-positive gram-negative rod that forms streptobacillary chains in some circumstances; it was therefore called the "ducreyi-like bacterium" (DLB). Distinguishing features of DLB are production of alpha-haemolysis on horse-blood agar, stimulation of growth by a microaerophilic atmosphere and by a factor produced by Staphylococcus aureus, a strongly positive porphyrin test, and a remarkable ability to undergo autolysis. DLB had a guanine + cytosine value of c. 50 mole% but it cannot be classified, even at the genus level, until more taxonomic data are obtained.

  9. A bacterium that degrades and assimilates poly(ethylene terephthalate).

    Science.gov (United States)

    Yoshida, Shosuke; Hiraga, Kazumi; Takehana, Toshihiko; Taniguchi, Ikuo; Yamaji, Hironao; Maeda, Yasuhito; Toyohara, Kiyotsuna; Miyamoto, Kenji; Kimura, Yoshiharu; Oda, Kohei

    2016-03-11

    Poly(ethylene terephthalate) (PET) is used extensively worldwide in plastic products, and its accumulation in the environment has become a global concern. Because the ability to enzymatically degrade PET has been thought to be limited to a few fungal species, biodegradation is not yet a viable remediation or recycling strategy. By screening natural microbial communities exposed to PET in the environment, we isolated a novel bacterium, Ideonella sakaiensis 201-F6, that is able to use PET as its major energy and carbon source. When grown on PET, this strain produces two enzymes capable of hydrolyzing PET and the reaction intermediate, mono(2-hydroxyethyl) terephthalic acid. Both enzymes are required to enzymatically convert PET efficiently into its two environmentally benign monomers, terephthalic acid and ethylene glycol.

  10. Genome analysis of the Anerobic Thermohalophilic bacterium Halothermothrix orenii

    Energy Technology Data Exchange (ETDEWEB)

    Mavromatis, Konstantinos; Ivanova, Natalia; Anderson, Iain; Lykidis, Athanasios; Hooper, Sean D.; Sun, Hui; Kunin, Victor; Lapidus, Alla; Hugenholtz, Philip; Patel, Bharat; Kyrpides, Nikos C.

    2008-11-03

    Halothermothirx orenii is a strictly anaerobic thermohalophilic bacterium isolated from sediment of a Tunisian salt lake. It belongs to the order Halanaerobiales in the phylum Firmicutes. The complete sequence revealed that the genome consists of one circular chromosome of 2578146 bps encoding 2451 predicted genes. This is the first genome sequence of an organism belonging to the Haloanaerobiales. Features of both Gram positive and Gram negative bacteria were identified with the presence of both a sporulating mechanism typical of Firmicutes and a characteristic Gram negative lipopolysaccharide being the most prominent. Protein sequence analyses and metabolic reconstruction reveal a unique combination of strategies for thermophilic and halophilic adaptation. H. orenii can serve as a model organism for the study of the evolution of the Gram negative phenotype as well as the adaptation under thermohalophilic conditions and the development of biotechnological applications under conditions that require high temperatures and high salt concentrations.

  11. Isolation of a bacterium that reductively dechlorinates tetrachloroethene to ethene

    Energy Technology Data Exchange (ETDEWEB)

    Maymo-Gatell, X.; Chien, Yueh-tyng; Zinder, S.H. [Cornell Univ., Ithaca, NY (United States)] [and others

    1997-06-06

    Tetrachloroethene is a prominent groundwater pollutant that can be reductively dechlorinated by mixed anaerobic microbial populations to the nontoxic product ethene. Strain 195, a coccoid bacterium that dechlorinates tetrachlorethene to ethene, was isolated and characterized. Growth of strain 195 with H{sub 2} and tetrachloroethene as the electron donor and acceptor pair required extracts from mixed microbial cultures. Growth of strain 195 was resistant to ampicillin and vancomycin; its cell wall did not react with a peptidoglycan-specific lectin and its ultrastructure resembled S-layers of Archaea. Analysis of the 16S ribosomal DNA sequence of strain 195 indicated that it is a eubacterium without close affiliation to any known groups. 24 refs., 4 figs., 1 tab.

  12. Erwinia amylovora loop-mediated isothermal amplification (LAMP) assay for rapid pathogen detection and on-site diagnosis of fire blight.

    Science.gov (United States)

    Bühlmann, Andreas; Pothier, Joël F; Rezzonico, Fabio; Smits, Theo H M; Andreou, Michael; Boonham, Neil; Duffy, Brion; Frey, Jürg E

    2013-03-01

    Several molecular methods have been developed for the detection of Erwinia amylovora, the causal agent of fire blight in pear and apple, but none are truly applicable for on-site use in the field. We developed a fast, reliable and field applicable detection method using a novel target on the E. amylovora chromosome that we identified by applying a comparative genomic pipeline. The target coding sequences (CDSs) are both uniquely specific for and all-inclusive of E. amylovora genotypes. This avoids potential false negatives that can occur with most commonly used methods based on amplification of plasmid gene targets, which can vary among strains. Loop-mediated isothermal AMPlification (LAMP) with OptiGene Genie II chemistry and instrumentation proved to be an exceptionally rapid (under 15 min) and robust method for detecting E. amylovora in orchards, as well as simple to use in the plant diagnostic laboratory. Comparative validation results using plant samples from inoculated greenhouse trials and from natural field infections (of regional and temporal diverse origin) showed that our LAMP had an equivalent or greater performance regarding sensitivity, specificity, speed and simplicity than real-time PCR (TaqMan), other LAMP assays, immunoassays and plating, demonstrating its utility for routine testing.

  13. Phylogenetic analysis of PR genes in some pome fruit species with the emphasis on transcriptional analysis and ROS response under Erwinia amylovora inoculation in apple.

    Science.gov (United States)

    Hassani, Maryam; Salami, Seyed Alireza; Nasiri, Jaber; Abdollahi, Hamid; Ghahremani, Zahra

    2016-02-01

    Attempts were made to identify eight pathogenesis related (PR) genes (i.e., PR-1a, PR3-ch1, PR3-Ch2, PR3-Ch3, PR3-Ch4, PR3-Ch5, PR-5 and PR-8) from 27 genotypes of apple, quince and pear, which are induced in response to inoculation with the pathogen Erwinia amylovora, the causal agent of fire blight. Totally, 32 PR genes of different families were obtained, excepting PR3-Ch2 (amplified only in apple) and PR3-Ch4 (amplified only in apple and pear), the others were successfully amplified in all the genotypes of apple, quince and pear. Evolutionary, the genes of each family exhibited significant homology with each other, as the corresponded phylogenetic neighbor-joining-based dendrograms were taken into consideration. Meanwhile, according to the expression assay, it was deduced that the pathogen activity can significantly affect the expression levels of some selected PR genes of PR3-Ch2, PR3-Ch4, PR3-Ch5 and particularly Cat I in both resistant (MM-111) and semi-susceptible (MM-106) apple rootstocks. Lastly, it was concluded that the pathogen E. amylovora is able to stimulate ROS response, particularly using generation of hydrogen peroxide (H2O2) in both aforementioned apple rootstock.

  14. Exploring new roles for the rpoS gene in the survival and virulence of the fire blight pathogen Erwinia amylovora.

    Science.gov (United States)

    Santander, Ricardo D; Monte-Serrano, Mercedes; Rodríguez-Herva, José J; López-Solanilla, Emilia; Rodríguez-Palenzuela, Pablo; Biosca, Elena G

    2014-12-01

    Erwinia amylovora causes fire blight in economically important plants of the family Rosaceae. This bacterial pathogen spends part of its life cycle coping with starvation and other fluctuating environmental conditions. In many Gram-negative bacteria, starvation and other stress responses are regulated by the sigma factor RpoS. We obtained an E. amylovora rpoS mutant to explore the role of this gene in starvation responses and its potential implication in other processes not yet studied in this pathogen. Results showed that E. amylovora needs rpoS to develop normal starvation survival and viable but nonculturable (VBNC) responses. Furthermore, this gene contributed to stationary phase cross-protection against oxidative, osmotic, and acid stresses and was essential for cross-protection against heat shock, but nonessential against acid shock. RpoS also mediated regulation of motility, exopolysaccharide synthesis, and virulence in immature loquats, but not in pear plantlets, and contributed to E. amylovora survival in nonhost tissues during incompatible interactions. Our results reveal some unique roles for the rpoS gene in E. amylovora and provide new knowledge on the regulation of different processes related to its ecology, including survival in different environments and virulence in immature fruits.

  15. T3SS-dependent differential modulations of the jasmonic acid pathway in susceptible and resistant genotypes of Malus spp. challenged with Erwinia amylovora.

    Science.gov (United States)

    Dugé De Bernonville, Thomas; Gaucher, Matthieu; Flors, Victor; Gaillard, Sylvain; Paulin, Jean-Pierre; Dat, James F; Brisset, Marie-Noëlle

    2012-06-01

    Fire blight is a bacterial disease of Maloideae caused by Erwinia amylovora (Ea). This necrogenic enterobacterium uses a type III secretion system (T3SS) to inject type III effectors into the plant cells to cause disease on its susceptible hosts, including economically important crops like apple and pear. The expressions of marker genes of the salicylic acid (SA) and jasmonic acid (JA) defense regulation pathways were monitored by RT-qPCR in leaves of two apple genotypes, one susceptible and one resistant, challenged with a wild type strain, a T3SS-deficient strain or water. The transcriptional data taken together with hormone level measurements indicated that the SA pathway was similarly induced in both apple genotypes during infection by Ea. On the contrary, the data clearly showed a strong T3SS-dependent down-regulation of the JA pathway in leaves of the susceptible genotype but not in those of the resistant one. Accordingly, methyl-jasmonate treated susceptible plants displayed an increased resistance to Ea. Bacterial mutant analysis indicated that JA manipulation by Ea mainly relies on the type III effector DspA/E. Taken together, our data suggest that the T3SS-dependent down-regulation of the JA pathway is a critical step in the infection process of Malus spp. by Ea.

  16. The tail-associated depolymerase of Erwinia amylovora phage L1 mediates host cell adsorption and enzymatic capsule removal, which can enhance infection by other phage.

    Science.gov (United States)

    Born, Yannick; Fieseler, Lars; Klumpp, Jochen; Eugster, Marcel R; Zurfluh, Katrin; Duffy, Brion; Loessner, Martin J

    2014-07-01

    The depolymerase enzyme (DpoL1) encoded by the T7-like phage L1 efficiently degrades amylovoran, an important virulence factor and major component of the extracellular polysaccharide (EPS) of its host, the plant pathogen Erwinia amylovora. Mass spectrometry analysis of hydrolysed EPS revealed that DpoL1 cleaves the galactose-containing backbone of amylovoran. The enzyme is most active at pH 6 and 50°C, and features a modular architecture. Removal of 180 N-terminal amino acids was shown not to affect enzyme activity. The C-terminus harbours the hydrolase activity, while the N-terminal domain links the enzyme to the phage particle. Electron microscopy demonstrated that DpoL1-specific antibodies cross-link phage particles at their tails, either lateral or frontal, and immunogold staining confirmed that DpoL1 is located at the tail spikes. Exposure of high-level EPS-producing Er. amylovora strain CFBP1430 to recombinant DpoL1 dramatically increased sensitivity to the Dpo-negative phage Y2, which was not the case for EPS-negative mutants or low-level EPS-producing Er. amylovora. Our findings indicate that enhanced phage susceptibility is based on enzymatic removal of the EPS capsule, normally a physical barrier to Y2 infection, and that use of DpoL1 together with the broad host range, virulent phage Y2 represents an attractive combination for biocontrol of fire blight.

  17. Discovery of plant phenolic compounds that act as type III secretion system inhibitors or inducers of the fire blight pathogen, Erwinia amylovora.

    Science.gov (United States)

    Khokhani, Devanshi; Zhang, Chengfang; Li, Yan; Wang, Qi; Zeng, Quan; Yamazaki, Akihiro; Hutchins, William; Zhou, Shan-Shan; Chen, Xin; Yang, Ching-Hong

    2013-09-01

    Erwinia amylovora causes a devastating disease called fire blight in rosaceous plants. The type III secretion system (T3SS) is one of the important virulence factors utilized by E. amylovora in order to successfully infect its hosts. By using a green fluorescent protein (GFP) reporter construct combined with a high-throughput flow cytometry assay, a library of phenolic compounds and their derivatives was studied for their ability to alter the expression of the T3SS. Based on the effectiveness of the compounds on the expression of the T3SS pilus, the T3SS inhibitors 4-methoxy-cinnamic acid (TMCA) and benzoic acid (BA) and one T3SS inducer, trans-2-(4-hydroxyphenyl)-ethenylsulfonate (EHPES), were chosen for further study. Both the T3SS inhibitors (TMCA and BA) and the T3SS inducer (EHPES) were found to alter the expression of T3SS through the HrpS-HrpL pathway. Additionally, TMCA altered T3SS expression through the rsmBEa-RsmAEa system. Finally, we found that TMCA and BA weakened the hypersensitive response (HR) in tobacco by suppressing the T3SS of E. amylovora. In our study, we identified phenolic compounds that specifically targeted the T3SS. The T3SS inhibitor may offer an alternative approach to antimicrobial therapy by targeting virulence factors of bacterial pathogens.

  18. The Erwinia amylovora PhoPQ system is involved in resistance to antimicrobial peptide and suppresses gene expression of two novel type III secretion systems.

    Science.gov (United States)

    Nakka, Sridevi; Qi, Mingsheng; Zhao, Youfu

    2010-10-20

    The PhoPQ system is a pleiotropic two-component signal transduction system that controls many pathogenic properties in several mammalian and plant pathogens. Three different cues have been demonstrated to activate the PhoPQ system including a mild acidic pH, antimicrobial peptides, and low Mg(2+). In this study, our results showed that phoPQ mutants were more resistant to strong acidic conditions (pH 4.5 or 5) than that of the wild-type (WT) strain, suggesting that this system in Erwinia amylovora may negatively regulate acid resistance gene expression. Furthermore, the PhoPQ system negatively regulated gene expression of two novel type III secretion systems in E. amylovora. These results are in contrast to those reported for the PhoPQ system in Salmonella and Xanthomonas, where it positively regulates type III secretion system and acid resistance. In addition, survival of phoPQ mutants was about 10-fold lower than that of WT when treated with cecropin A at pH 5.5, suggesting that the PhoPQ system renders the pathogen more resistant to cecropin A.

  19. The outer membrane protein TolC is required for phytoalexin resistance and virulence of the fire blight pathogen Erwinia amylovora.

    Science.gov (United States)

    Al-Karablieh, Nehaya; Weingart, Helge; Ullrich, Matthias S

    2009-07-01

    Erwinia amylovora causes fire blight on several plant species such as apple and pear, which produce diverse phytoalexins as defence mechanisms. An evolutionary successful pathogen thus must develop resistance mechanisms towards these toxic compounds. The E. amylovora outer membrane protein, TolC, might mediate phytoalexin resistance through its interaction with the multidrug efflux pump, AcrAB. To prove this, a tolC mutant and an acrB/tolC double mutant were constructed. The minimal inhibitory concentrations of diverse antimicrobials and phytoalexins were determined for these mutants and compared with that of a previously generated acrB mutant. The tolC and arcB/tolC mutants were considerably more susceptible than the wild type but showed similar levels as the acrB mutant. The results clearly indicated that neither TolC nor AcrAB significantly interacted with other transport systems during the efflux of the tested toxic compounds. Survival and virulence assays on inoculated apple plants showed that pathogenicity and the ability of E. amylovora to colonize plant tissue were equally impaired by mutations of tolC and acrB/tolC. Our results allowed the conclusion that TolC plays an important role as a virulence and fitness factor of E. amylovora by mediating resistance towards phytoalexins through its exclusive interaction with AcrAB.

  20. In-vitro antibacterial activities of the essential oils of aromatic plants against Erwinia herbicola (Lohnis and pseudomonas putida (Kris Hamilton

    Directory of Open Access Journals (Sweden)

    Pandey Abhay K.

    2012-01-01

    Full Text Available This study was designed to examine in vitro antibacterial activities of essential oils extracted from 53 aromatic plants of Gorakhpur Division (UP, INDIA for the control of two phytopathogenic bacteria namely Erwinia herbicola and Pseudomonas putida causing several post-harvest diseases in fruits and vegetables. Out of 53 oils screened, 8 oils such as Chenopodium ambrosioides, Citrus aurantium, Clausena pentaphylla, Hyptis suaveolens, Lippia alba, Mentha arvensis, Ocimum sanctum and Vitex negundo completely inhibited the growth of test bacteria. Furthermore MIC & MBC values of C. ambrosioides oil were least for Erw. herbicola (0.25 & 2.0 μl/ml and Ps. putida (0.12 & 1.0 μl/ml respectively than other 7 oils as well as Agromycin and Streptomycin drugs used in current study. GC and GC-MS analysis of Chenopodium oil revealed presence of 125 major and minor compounds, out of them, 14 compounds were recognized. The findings concluded that Chenopodium oil may be regarded as safe antibacterial agent for the management of post-harvest diseases of fruits and vegetables.

  1. Characterisation of the stbD/E toxin-antitoxin system of pEP36, a plasmid of the plant pathogen Erwinia pyrifoliae.

    Science.gov (United States)

    Unterholzner, Simon J; Hailer, Barbara; Poppenberger, Brigitte; Rozhon, Wilfried

    2013-09-01

    pEP36 is a plasmid ubiquitously present in Erwinia pyrifoliae, a pathogen which causes black stem blight of Asian pear. pEP36 is highly stable in its host, even in the absence of selective pressure. The plasmid is closely related to pEA29, which is widespread in E. amylovora, the causative agent of fire blight of apple and pear trees. Here we report that pEP36 possesses a functional hybrid toxin-antitoxin module, stbD/E(pEP36), with the toxin showing homology to the RelE/ParE proteins and the antidote belonging to the Phd/YefM antitoxin family. Bacteria expressing the StbE(pEP36) toxin arrest cell growth and enter a viable but non-culturable stage. However, they maintain their typical cell length and do not show filamentation. Pulse-chase experiments revealed that StbE(pEP36) acts as a global inhibitor of protein synthesis while it does not interfere with DNA and RNA synthesis. The StbD(pEP36) antitoxin is capable of neutralising StbE(pEP36) toxicity. Additional experiments show that the stbD/E(pEP36) module can stabilise plasmids at least 20-fold. Thus the toxin-antitoxin system may contribute to the remarkable stability of pEP36.

  2. The pathogeny occurrence pattern and control of Erwinia Jujubovora%枣缩果病病原发病规律及防治

    Institute of Scientific and Technical Information of China (English)

    任士福; 张彦婷

    2001-01-01

    @@ 枣缩果病(Erwinia Jujubovora)又名铁皮病、黑腐病,是目前枣树生产中最重要的果实病害.该病在枣果白熟期开始发病,病果果肉变褐,味变苦,造成提前落果,导致产量和品质下降.从20世纪80年代初开始,该病为害日趋严重,其发病快而集中,常常暴发流行.河北省每年由于该病导致病果产量损失30%~80%,有的地方甚至出现绝收.该病在河南、山东、山西、内蒙古、广西等省区也有发生,有的地方发生的也相当严重,已成为当前大枣生产中亟待解决的重大问题.现介绍河北农业大学最新研究成果,以指导枣产区的生产.

  3. Pathogenicity and Biological Characters of Erwinia chyrsanthemi in Yunnan Province%云南水稻细菌性基腐病的致病性及病原生物学特性研究

    Institute of Scientific and Technical Information of China (English)

    周惠萍; 范静华; 徐自怀; 陈建斌

    2004-01-01

    通过对云南省水稻细菌性基腐病菌进行细菌学性状、致病性和温度测定,结果表明:该病原菌属于菊欧氏杆菌玉米致病变种(Erwinia chyrsanthemi pv.zeae),其生长适宜温度范围28~36 ℃,其中以32 ℃为最适,最低温度为10 ℃,最高温度42 ℃,致死温度53 ℃,10 min.

  4. The Genomic DNA Library Construction of Erwinia Carotovora CXJZ95-198%欧文氏杆菌CXJZ95-198基因组文库的构建

    Institute of Scientific and Technical Information of China (English)

    张运雄; 刘正初

    2006-01-01

    采用改良鸟枪法构建了草本纤维精制高效菌种Erwinia carotovora CXJZ95-198的基因组文库,结合透明圈法,筛选到了8个甘露聚糖酶基因阳性克隆,并采用PCR方法对它们进行了分析鉴定,结果表明它们含有同一个甘露聚糖酶基因.

  5. Study on resistance of Chinese cabbage to Erwinia carotovora subsp.carotovora by exogenous 24-Epibrassinolide treatment%表油菜素内酯诱导大白菜抗软腐病研究

    Institute of Scientific and Technical Information of China (English)

    屈淑平; 张灵宇; 崔崇士

    2009-01-01

    To ascertain the induced resistance to Erwinia carotovora subsp. carotovora by 24-Epibra-ssinolide and its physiological mechanism, Chinese cabbage was used to study the effects of 24-Epibra-ssinolide on its physiological and biochemical characteristics with foliar spray. The results showed that 24-Epibrassinolide decreased disease indexes of Chinese cabbage seedlings by 25.8%, increased H_2O_2 accumulation, decreased MDA contents, increased catalase (CAT) activities, peroxidase (POD) activities trend, increased slowly in early phase, and decreased in later stage. Erwinia carotovora subp. carotovora infection resulted in increased levels of H_2O_2 and MDA when plants were sprayed with EBR, but the MDA content was lower than the control treatment. Foliar spraying with EBR increased CAT activities, and POD activities reduced than the control treatment. EBR promoted the growth of Chinese cabbage seedling, this increasing appeared even after Erwinia carotovora subsp. carotovora infection. EBR kept the metabolism of active oxygen species at a balance state and increased the resistance to Erwinia carotovora subsp. carotovora in Chinese cabbage.%试验采用叶面喷施的方法,研究了24-表油菜素内酯(24-Epibrassinolide,EBR)对大白菜软腐病的诱抗作用及其生理机制.结果表明,外源EBR处理大白菜幼苗病情指教明显比对照降低,下降了25.8%;喷施EBR提高了大白菜叶片中H_2O_2含量,使丙二醛(Malondialdehyde,MDA)含量下降,提高了过氧化氢酶(Catalase,CAT)活性,过氧化物酶(Peroxidase,POD)活性先缓慢升高然后下降;接种软腐病菌后,EBR使H_2O_2和MDA含量缓慢增加,但EBR处理MDA含量低于对照处理,显著提高了CAT活性,POD活性与对照相比下降,呈现缓慢升高趋势;EBR处理明显促进幼苗的生长,即使在接种软腐病菌期间,植株的长势也比对照强.说明EBR通过调节植物体内氧的代谢平衡,增强对软腐病的抗性.

  6. Portable hyperspectral fluorescence imaging system for detection of biofilms on stainless steel surfaces

    Science.gov (United States)

    Jun, Won; Lee, Kangjin; Millner, Patricia; Sharma, Manan; Chao, Kuanglin; Kim, Moon S.

    2008-04-01

    A rapid nondestructive technology is needed to detect bacterial contamination on the surfaces of food processing equipment to reduce public health risks. A portable hyperspectral fluorescence imaging system was used to evaluate potential detection of microbial biofilm on stainless steel typically used in the manufacture of food processing equipment. Stainless steel coupons were immersed in bacterium cultures, such as E. coli, Pseudomonas pertucinogena, Erwinia chrysanthemi, and Listeria innocula. Following a 1-week exposure, biofilm formations were assessed using fluorescence imaging. In addition, the effects on biofilm formation from both tryptic soy broth (TSB) and M9 medium with casamino acids (M9C) were examined. TSB grown cells enhance biofilm production compared with M9C-grown cells. Hyperspectral fluorescence images of the biofilm samples, in response to ultraviolet-A (320 to 400 nm) excitation, were acquired from approximately 416 to 700 nm. Visual evaluation of individual images at emission peak wavelengths in the blue revealed the most contrast between biofilms and stainless steel coupons. Two-band ratios compared with the single-band images increased the contrast between the biofilm forming area and stainless steel coupon surfaces. The 444/588 nm ratio images exhibited the greatest contrast between the biofilm formations and stainless coupon surfaces.

  7. The lipopolysaccharide of a chloridazon-degrading bacterium.

    Science.gov (United States)

    Weisshaar, R; Lingens, F

    1983-12-01

    Lipopolysaccharide of a chloridazon-degrading bacterium was obtained by a two-stage extraction procedure with phenol/EDTA in a yield of 0.3% of dried bacteria. The carbohydrate moiety consisted of heptose, 3-deoxyoctulosonic acid and D-glucose in a molar ratio of 1:2:2 X 3. Lipid A was composed of 1 mol 2,3-diamino-2,3-dideoxy-D-glucose, 2 mol amide-bound and 2.6 mol ester-bound fatty acids/mol. Amide-bound fatty acids were 3-hydroxydodecanoic acid and 3-hydroxyhexadecanoic acid; dodecanoic acid and R-(-)-3-hydroxydodec-5-cis-enoic acid were found to be present in ester linkage. Under conditions of acidic hydrolysis, the latter was converted into the cis and trans isomers of 5-hexyltetrahydrofuran-2-acetic acid. Dodecanoic acid was demonstrated to be linked with the hydroxy groups of the amide-bound fatty acids. The taxonomic significance of these results, especially the demonstration of 2,3-diamino-2, 3-dideoxy-D-glucose, is discussed.

  8. Presence of an unusual methanogenic bacterium in coal gasification waste.

    Science.gov (United States)

    Tomei, F A; Rouse, D; Maki, J S; Mitchell, R

    1988-12-01

    Methanogenic bacteria growing on a pilot-scale, anaerobic filter processing coal gasification waste were enriched in a mineral salts medium containing hydrogen and acetate as potential energy sources. Transfer of the enrichments to methanol medium resulted in the initial growth of a strain of Methanosarcina barkeri, but eventually small cocci became dominant. The cocci growing on methanol produced methane and exhibited the typical fluorescence of methanogenic bacteria. They grew in the presence of the cell wall synthesis-inhibiting antibiotics d-cycloserine, fosfomycin, penicillin G, and vancomycin as well as in the presence of kanamycin, an inhibitor of protein synthesis in eubacteria. The optimal growth temperature was 37 degrees C, and the doubling time was 7.5 h. The strain lysed after reaching stationary phase. The bacterium grew poorly with hydrogen as the energy source and failed to grow on acetate. Morphologically, the coccus shared similarities with Methanosarcina sp. Cells were 1 mum wide, exhibited the typical thick cell wall and cross-wall formation, and formed tetrads. Packets and cysts were not formed.

  9. Tracing the run-flip motion of an individual bacterium

    Science.gov (United States)

    Liu, Bin; Morse, Michael; Tang, Jay; Powers, Thomas; Breuer, Kenneth S.

    2012-11-01

    We have developed a digital 3D tracking microscope in which the microscope stage follows the motion of an individual motile microorganism so that the target remains focused at the center of the view-field. The tracking mechanism is achieved by a high-speed feedback control through real-time image analysis and the trace of the microorganism is recorded with submicron accuracy. We apply this tracking microscope to a study of the motion of an individual Caulobacter crescentus, a bacterium that moves up to 100 microns (or 50 body lengths) per second and reverses its direction of motion occasionally by switching the rotation direction of its single helical flagellum. By tracking the motion of a single cell over many seconds, we show how a flip event occurs with submicron resolution and how the speed of a single cell varies over time and with the rotational rate of the flagellum. We also present statistics for the run-reverse dynamics of an ensemble of cells.

  10. Novel Trypanosomatid-Bacterium Association: Evolution of Endosymbiosis in Action

    Directory of Open Access Journals (Sweden)

    Alexei Y. Kostygov

    2016-03-01

    Full Text Available We describe a novel symbiotic association between a kinetoplastid protist, Novymonas esmeraldas gen. nov., sp. nov., and an intracytoplasmic bacterium, “Candidatus Pandoraea novymonadis” sp. nov., discovered as a result of a broad-scale survey of insect trypanosomatid biodiversity in Ecuador. We characterize this association by describing the morphology of both organisms, as well as their interactions, and by establishing their phylogenetic affinities. Importantly, neither partner is closely related to other known organisms previously implicated in eukaryote-bacterial symbiosis. This symbiotic association seems to be relatively recent, as the host does not exert a stringent control over the number of bacteria harbored in its cytoplasm. We argue that this unique relationship may represent a suitable model for studying the initial stages of establishment of endosymbiosis between a single-cellular eukaryote and a prokaryote. Based on phylogenetic analyses, Novymonas could be considered a proxy for the insect-only ancestor of the dixenous genus Leishmania and shed light on the origin of the two-host life cycle within the subfamily Leishmaniinae.

  11. Electromicrobiology of Dissimilatory Sulfur Reducing Bacterium Desulfuromonas acetexigens

    KAUST Repository

    Bin Bandar, Khaled

    2014-12-01

    Bioelectrochmical systems (BES) are engineered electrochemical devices that harness hidden chemical energy of the wastewater in to the form of electricity or hydrogen. Unique microbial communities enrich in these systems for oxidation of organic matter as well as transfer of resulted electron to anode, known them as “electricigens” communities. Exploring novel electricigenesis microbial communities in the nature and understanding their electromicrobiology is one the important aspect for BES systems scale up. Herein, we report first time the electricigenesis property of an anaerobic, fresh water sediment, sulfur reducing bacterium Desulfuromona acetexigens. The electrochemical behavior of D. acetexigens biofilms grown on graphite-rod electrodes in batch-fed mode under an applied potential was investigated with traditional electroanalytical tools, and correlate the electron transfer from biofilms to electrode with a model electricigen Geobacter sulfurreducens electrochemical behavior. Research findings suggest that D. acetexigens has the ability to use electrode as electron acceptor in BES systems through establishing the direct contact with anode by expressing the membrane bound redox proteins, but not due to the secretion of soluble redox mediators. Preliminary results revealed that D. acetexigens express three distinct redox proteins in their membranes for turnover of the electrons from biofilm to electrode, and the 4 whole electricigenesis process observed to be unique in the D. acetexigens compared to that of well-studied model organism G. sulfurreducens.

  12. Denitrification characteristics of a marine origin psychrophilic aerobic denitrifying bacterium

    Institute of Scientific and Technical Information of China (English)

    Haiyan Zheng; Ying Liu; Guangdong Sun; Xiyan Gao; Qingling Zhang; Zhipei Liu

    2011-01-01

    A psychrophilic aerobic denitrifying bacterium,strain S1-1,was isolated from a biological aerated filter conducted for treatment of recirculating water in a marine aquaculture system.Strain S1-1 was preliminarily identified as Psychrobacter sp.based on the analysis of its 16S rRNA gene sequence,which showed 100% sequence similarity to that of Psychrobacter sp.TSBY-70.Strain S 1-1 grew well either in high nitrate or high nitrite conditions with a removal of 100% nitrate or 63.50% nitrite,and the total nitrogen removal rates could reach to 46.48% and 31.89%,respectively.The results indicated that nitrate was mainly reduced in its logarithmic growth phase with a very low leve 1 accumulation of nitrite,suggesting that the aerobic denitrification process of strain S l-1 occurred mainly in this phase.The GC-MS results showed that N2O was formed as the major intermediate during the aerobic denitrifying process of strain S1-1.Finally,factors affecting the growth of strain Sl-1 and its aerobic denitrifying ability were also investigated.Results showed that the optimum aerobic denitrification conditions for strain S1-1 were sodium succinate as carbon source,C/N ratio15,salinity 10 g/L NaCl,incubation temperature 20℃ and initial pH 6.5.

  13. Pandoraea sp. RB-44, A Novel Quorum Sensing Soil Bacterium

    Directory of Open Access Journals (Sweden)

    Robson Ee Han-Jen

    2013-10-01

    Full Text Available Proteobacteria are known to communicate via signaling molecules and this process is known as quorum sensing. The most commonly studied quorum sensing molecules are N-acylhomoserine lactones (AHLs that consists of a homoserine lactone moiety and an N-acyl side chain with various chain lengths and degrees of saturation at the C-3 position. We have isolated a bacterium, RB-44, from a site which was formally a landfill dumping ground. Using matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF mass spectrometry analysis, this isolate was identified as a Pandoraea sp.which was then screened for AHL production using biosensors which indicated its quorum sensing properties. To identify the AHL profile of Pandoraea sp. RB-44, we used high resolution tandem mass spectrometry confirming that this isolate produced N-octanoylhomoserine lactone (C8-HSL. To the best of our knowledge, this is the first report that showed quorum sensing activity exhibited by Pandoraea sp. Our data add Pandoraea sp. to the growing number of bacteria that possess QS systems.

  14. Biochemical characterization and immobilization of Erwinia carotovoral-asparaginase in a microplate for high-throughput biosensing of l-asparagine.

    Science.gov (United States)

    Labrou, Nikolaos E; Muharram, Magdy Mohamed

    2016-10-01

    l-Asparaginases (l-ASNase, E.C. 3.5.1.1) catalyze the conversion of l-asparagine to l-aspartic acid and ammonia. In the present work, a new form of l-ASNase from a strain of Erwinia carotovora (EcaL-ASNase) was cloned, expressed in Escherichia coli as a soluble protein and characterized. The enzyme was purified to homogeneity by a single-step procedure comprising ion-exchange chromatography. The properties of the recombinant enzyme were investigated employing kinetic analysis and molecular modelling and the kinetic parameters (Km, kcat) were determined for a number of substrates. The enzyme was used to assemble a microplate-based biosensor that was used for the development of a simple assay for the determination of l-asparagine in biological samples. In this sensor, the enzyme was immobilized by crosslinking with glutaraldehyde and deposited into the well of a microplate in 96-well format. The sensing scheme was based on the colorimetric measurement of ammonia formation using the Nessler's reagent. This format is ideal for micro-volume applications and allows the use of the proposed biosensor in high-throughput applications for monitoring l-asparagine levels in serum and foods samples. Calibration curve was obtained for l-asparagine, with useful concentration range 10-200μΜ. The biosensor had a detection limit of 10μM for l-asparagine. The method's reproducibility was in the order of ±3-6% and l-asparagine mean recoveries were 101.5%.

  15. Conserved aspartate and lysine residues of RcsB are required for amylovoran biosynthesis, virulence, and DNA binding in Erwinia amylovora.

    Science.gov (United States)

    Ancona, Veronica; Chatnaparat, Tiyakhon; Zhao, Youfu

    2015-08-01

    In Erwinia amylovora, the Rcs phosphorelay system is essential for amylovoran production and virulence. To further understand the role of conserved aspartate residue (D56) in the phosphor receiver (PR) domain and lysine (K180) residue in the function domain of RcsB, amino acid substitutions of RcsB mutant alleles were generated by site-directed mutagenesis and complementation of various rcs mutants were performed. A D56E substitution of RcsB, which mimics the phosphorylation state of RcsB, complemented the rcsB mutant, resulting in increased amylovoran production and gene expression, reduced swarming motility, and restored pathogenicity. In contrast, D56N and K180A or K180Q substitutions of RcsB did not complement the rcsB mutant. Electrophoresis mobility shift assays showed that D56E, but not D56N, K180Q and K180A substitutions of RcsB bound to promoters of amsG and flhD, indicating that both D56 and K180 are required for DNA binding. Interestingly, the RcsBD56E allele could also complement rcsAB, rcsBC and rcsABCD mutants with restored virulence and increased amylovoran production, indicating that RcsB phosphorylation is essential for virulence of E. amylovora. In addition, mutations of T904 and A905, but not phosphorylation mimic mutation of D876 in the PR domain of RcsC, constitutively activate the Rcs system, suggesting that phosphor transfer is required for activating the Rcs system and indicating both A905 and T904 are required for the phosphatase activity of RcsC. Our results demonstrated that RcsB phosphorylation and dephosphorylation, phosphor transfer from RcsC are essential for the function of the Rcs system, and also suggested that constitutive activation of the Rcs system could reduce the fitness of E. amylovora.

  16. Control of fire blight (Erwinia amylovora on apple trees with trunk-injected plant resistance inducers and antibiotics and assessment of induction of pathogenesis-related protein genes

    Directory of Open Access Journals (Sweden)

    Srđan G. Aćimović

    2015-02-01

    Full Text Available Management of fire blight is complicated by limitations on use of antibiotics in agriculture, antibiotic resistance development, and limited efficacy of alternative control agents. Even though successful in control, preventive antibiotic sprays also affect non-target bacteria, aiding the selection for resistance which could ultimately be transferred to the pathogen Erwinia amylovora. Trunk injection is a target-precise pesticide delivery method that utilizes tree xylem to distribute injected compounds. Trunk injection could decrease antibiotic usage in the open environment and increase the effectiveness of compounds in fire blight control. In field experiments, after 1-2 apple tree injections of either streptomycin, potassium phosphites (PH or acibenzolar-S-methyl (ASM, significant reduction of blossom and shoot blight symptoms was observed compared to water- or non-injected control trees. Overall disease suppression with streptomycin was lower than typically observed following spray applications to flowers. Trunk injection of oxytetracycline resulted in excellent control of shoot blight severity, suggesting that injection is a superior delivery method for this antibiotic. Injection of both ASM and PH resulted in the significant induction of PR-1, PR-2 and PR-8 protein genes in apple leaves indicating induction of systemic acquired resistance (SAR under field conditions. The time separating SAR induction and fire blight symptom suppression indicated that various defensive compounds within the SAR response were synthesized and accumulated in the canopy. ASM and PH suppressed fire blight even after cessation of induced gene expression. With the development of injectable formulations and optimization of doses and injection schedules, the injection of protective compounds could serve as an effective option for fire blight control.

  17. AraC/XylS family stress response regulators Rob, SoxS, PliA, and OpiA in the fire blight pathogen Erwinia amylovora.

    Science.gov (United States)

    Pletzer, Daniel; Schweizer, Gabriel; Weingart, Helge

    2014-09-01

    Transcriptional regulators of the AraC/XylS family have been associated with multidrug resistance, organic solvent tolerance, oxidative stress, and virulence in clinically relevant enterobacteria. In the present study, we identified four homologous AraC/XylS regulators, Rob, SoxS, PliA, and OpiA, from the fire blight pathogen Erwinia amylovora Ea1189. Previous studies have shown that the regulators MarA, Rob, and SoxS from Escherichia coli mediate multiple-antibiotic resistance, primarily by upregulating the AcrAB-TolC efflux system. However, none of the four AraC/XylS regulators from E. amylovora was able to induce a multidrug resistance phenotype in the plant pathogen. Overexpression of rob led to a 2-fold increased expression of the acrA gene. However, the rob-overexpressing strain showed increased resistance to only a limited number of antibiotics. Furthermore, Rob was able to induce tolerance to organic solvents in E. amylovora by mechanisms other than efflux. We demonstrated that SoxS from E. amylovora is involved in superoxide resistance. A soxS-deficient mutant of Ea1189 was not able to grow on agar plates supplemented with the superoxide-generating agent paraquat. Furthermore, expression of soxS was induced by redox cycling agents. We identified two novel members of the AraC/XylS family in E. amylovora. PliA was highly upregulated during the early infection phase in apple rootstock and immature pear fruits. Multiple compounds were able to induce the expression of pliA, including apple leaf extracts, phenolic compounds, redox cycling agents, heavy metals, and decanoate. OpiA was shown to play a role in the regulation of osmotic and alkaline pH stress responses.

  18. Erwinia carotovora ssp. carotovora Infection Induced "Defense Lignin" Accumulation and Lignin Biosynthetic Gene Expression in Chinese Cabbage (Brassica rapa L. ssp. pekinensis)

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    Erwinia carotovora subsp. carotovora (Ecc) infects and causes soft rot disease in hundreds of crop species including vegetables, flowers and fruits. Lignin biosynthesis has been implicated in defensive reactions to injury and pathogen Infection in plants. In this work, variations of lignin content and gene expression in the molecular interaction between Chinese cabbage and Ecc were investigated. H2O2 accumulation and peroxidase activity were detected by 3, 3'-Dimethoxybenzidine staining at mocked and Ecc-inoculated sites of Chinese cabbage leafstalks. Klason lignin content in inoculated plants increased by about 7.84%, 40.37%, and 43.13% more than that of the mocked site at 12, 24 and 72 h after inoculation, respectively. Gas chromatography detected more p-coumaryl (H) and less coniferyl (G) and sinapyl (S)monolignins in leafstalks of Chinese cabbage. All three monomers increased in Ecc-infected leafstalks, and the Ecc-induced "defense lignin" were composed of more G and H monolignins, and less S monolignin. After searching the expressed sequence tags (EST) data of Chinese cabbage, 12 genes putatively encoding enzymes involved in lignin biosynthesis were selected to study their expression. All of these genes could be Induced by mock inoculation and Ecc infection, while the gene expression lasted for several more hours in the infected samples than in mocked and untreated plants. Our results indicated that "defense lignin" was different from the developmental lignin in composition; G and S monolignins were significantly induced in plants in response to the soft rot Ecc; thus, lignin biosynthesis was differentially regulated and played a role in plant response to the soft rot Ecc.

  19. Control of postharvest soft rot caused by Erwinia carotovora of vegetables by a strain of Bacillus amyloliquefaciens and its potential modes of action.

    Science.gov (United States)

    Zhao, Yancun; Li, Pengxia; Huang, Kaihong; Wang, Yuning; Hu, Huali; Sun, Ya

    2013-03-01

    Erwinia carotovora subsp. carotovora (Ecc), the causal agent of bacterial soft rot, is one of the destructive pathogens of postharvest vegetables. In this study, a bacterial isolate (BGP20) from the vegetable farm soil showed strong antagonistic activity against Ecc in vitro, and its twofold cell-free culture filtrate showed excellent biocontrol effect in controlling the postharvest bacterial soft rot of potatoes at 25 °C. The anti-Ecc metabolites produced by the isolate BGP20 had a high resistance to high temperature, UV-light and protease K. Based on the colonial morphology, cellular morphology, sporulation, and partial nucleotide sequences of 16S rRNA and gyrB gene, the isolate BGP20 was identified as Bacillus amyloliquefaciens subsp. plantarum. Further in vivo assays showed that the BGP20 cell culture was more effective in controlling the postharvest bacterial soft rot of green peppers and Chinese cabbages than its twofold cell-free culture filtrate. In contrast, the biocontrol effect and safety of the BGP20 cell culture were very poor on potatoes. In the wounds of potatoes treated with both the antagonist BGP20 and the pathogen Ecc, the viable count of Ecc was 31,746 times that of BGP20 at 48 h of incubation at 25 °C. But in the wounds of green peppers, the viable count of BGP20 increased 182.3 times within 48 h, and that of Ecc increased only 51.3 %. In addition, the treatment with both BGP20 and Ecc induced higher activity of phenylalanine ammonia-lyase (PAL) than others in potatoes. But the same treatment did not induce an increase of PAL activity in green peppers. In conclusion, the present study demonstrated that the isolate BGP20 is a promising candidate in biological control of postharvest bacterial soft rot of vegetables, but its main mode of action is different among various vegetables.

  20. IN SITU RT-PCR WITH A SULFATE-REDUCING BACTERIUM ISOLATED FROM SEAGRASS ROOTS

    Science.gov (United States)

    Bacteria considered to be obligate anaerobes internally colonize roots of the submerged macrophyte Halodule wrightii. A sulfate reducing bacterium, Summer lac 1, was isolated on lactate from H. wrightii roots. The isolate has physiological characteristics typical of Desulfovibri...

  1. 化学抑制剂Woodward′s Reagent K对来源于Erwinia rhapontici NX 5蔗糖异构酶的抑制动力学%Inhibition kinetics of sucrose isomerase from Erwinia rhapontici NX-5 by Woodward′s Reagent K

    Institute of Scientific and Technical Information of China (English)

    王彦媛; 李莎; 姚忠; 徐虹

    2014-01-01

    从重组大肠杆菌E�coli BL21( pET22b palⅠ)中纯化得到来源于Erwinia rhapontici NX 5的蔗糖异构酶(sucrose isomerase,SIase,EC 5�4�99�11),以纯酶为对象考察其酶活力抑制动力学。结果表明:SIase 纯比酶活1512�77 U/mg,动力学常数 Km=260 mmol/L,Vmax=39�41μmol/(L·s)。以化学抑制剂 Woodward′s Reagent K (WRK)对重组蔗糖异构酶进行抑制反应,反应体系中随着WRK浓度的升高,SIase与底物蔗糖的亲和力常数Km增大,最大反应速度Vmax在一定范围内保持稳定。通过对SIase的抑制动力学分析可得到,WRK对SIase的抑制类型为可逆的竞争性抑制,这可能与WRK与蔗糖的结构类似,与可竞争性的结合SIase的活性中心有关。%Sucrose isomerase ( SIase,EC5�4�99�11) from Erwinia rhapontici NX⁃5 was purified from the extract of recombinant E�coli BL21 ( pET22b⁃palⅠ) culture,and the inhibition kinetics of the pure SIase was studied with chemical inhibitor Woodward′s Reagent K( WRK)�Results show that SIase had the high specific activity of 1 512�77 U/mg,as well as the Michaelis⁃Menten constants of Km=260 mmol/L and Vmax=39�41 μmol/( L·s)�Km increased as the concentration of inhibitor increased,but Vmax kept stable within limits�The inhibition of SIase by WRK was reversible and competitive, probably caused by the similar structure of WRK and sucrose.

  2. Effect of alginic acid decomposing bacterium on the growth of Laminaria japonica (Phaeophyceae)

    Institute of Scientific and Technical Information of China (English)

    WANG You; TANG Xue-xi; YANG Zhen; YU Zhi-ming

    2006-01-01

    We collected the diseased blades of Laminaria japonica from Yantai Sea Farm from October to December 2002, and the alginic acid decomposing bacterium on the diseased blade was isolated and purified, and was identified as Alteromonas espejiana. This bacterium was applied as the causative pathogen to infect the blades of L. japonica under laboratory conditions. The aim of the present study was to identify the effects of the bacterium on the growth of L. japonica, and to find the possibly effective mechanism. Results showed that: (1)The blades of L.japonica exhibited symptoms of lesion,bleaching and deterioration when infected by the bacterium,and their growth and photosynthesis were dramatically suppressed. At the same time, the reactive oxygen species (ROS) generation enhanced obviously, and the relative membrane permeability increased significantly. The contents of malonaldehyde (MDA) and free fatty acid in the microsomol membrane greatly elevated, but the phospholipid content decreased. Result suggested an obvious peroxidation and deesterrification in the blades of L. japonica when infected by the bacterium. (2) The simultaneous assay on the antioxidant enzyme activities demonstrated that superoxide dismutase (SOD) and catalase (CAT) increased greatly when infected by the bacterium, but glutathione peroxidase (Gpx) and ascorbate peroxidase (APX) did not exhibit active responses to the bacterium throughout the experiment. (3) The histomorphological observations gave a distinctive evidence of the severity of the lesions as well as the relative abundance in the bacterial population on the blades after infection. The bacterium firstly invaded into the endodermis of L. japonica and gathered around there, and then resulted in the membrane damage, cells corruption and ultimately, the death of L.japonica.

  3. Endohyphal bacterium enhances production of indole-3-acetic acid by a foliar fungal endophyte.

    Science.gov (United States)

    Hoffman, Michele T; Gunatilaka, Malkanthi K; Wijeratne, Kithsiri; Gunatilaka, Leslie; Arnold, A Elizabeth

    2013-01-01

    Numerous plant pathogens, rhizosphere symbionts, and endophytic bacteria and yeasts produce the important phytohormone indole-3-acetic acid (IAA), often with profound effects on host plants. However, to date IAA production has not been documented among foliar endophytes -- the diverse guild of primarily filamentous Ascomycota that live within healthy, above-ground tissues of all plant species studied thus far. Recently bacteria that live within hyphae of endophytes (endohyphal bacteria) have been detected, but their effects have not been studied previously. Here we show not only that IAA is produced in vitro by a foliar endophyte (here identified as Pestalotiopsis aff. neglecta, Xylariales), but that IAA production is enhanced significantly when the endophyte hosts an endohyphal bacterium (here identified as Luteibacter sp., Xanthomonadales). Both the endophyte and the endophyte/bacterium complex appear to rely on an L-tryptophan dependent pathway for IAA synthesis. The bacterium can be isolated from the fungus when the symbiotic complex is cultivated at 36°C. In pure culture the bacterium does not produce IAA. Culture filtrate from the endophyte-bacterium complex significantly enhances growth of tomato in vitro relative to controls and to filtrate from the endophyte alone. Together these results speak to a facultative symbiosis between an endophyte and endohyphal bacterium that strongly influences IAA production, providing a new framework in which to explore endophyte-plant interactions.

  4. 进境韩国兰花细菌性褐腐病菌的分离与鉴定%Isolation and Identification of Erwinia cypripedii from Imported Korea Cymbidium

    Institute of Scientific and Technical Information of China (English)

    厉艳; 王英超; 尼秀媚; 甘琴华; 封立平

    2012-01-01

    [目的]鉴定进境韩国兰花细菌性褐腐病菌.[方法]从韩国进境的大花蕙兰植株病变叶片中分离到细菌菌株,并通过形态鉴定、培养特征、生理生化及16S rDNA序列分析和致病性测定对其进行了鉴定.[结果]确认该病菌为兰花细菌性褐腐病菌(Erwinia cypripedii),这是我国首次从进境兰花中检出该病害.[结论]为我国兰花细菌性褐腐病菌的预防及控制奠定了基础.%The aim was to identify Erwinia cypripedii from imported Korea Cymbidium. [ Method] The pathogenic bacteria was i-solated from imported Korea Cymbidium leaves, and then it was identified by morphological identification, cultural characteristics, physiological and biochemical reactions, 16S rDNA gene sequences analysis and pathogenicity test. [Result] The isolated strain was identified as E. cypripedii, and it was the first interception in China. [ Conclusion ] The research result lays the foundation for the prevention and control of E. cypripedii in China.

  5. Metabolic evolution of a deep-branching hyperthermophilic chemoautotrophic bacterium.

    Directory of Open Access Journals (Sweden)

    Rogier Braakman

    Full Text Available Aquifex aeolicus is a deep-branching hyperthermophilic chemoautotrophic bacterium restricted to hydrothermal vents and hot springs. These characteristics make it an excellent model system for studying the early evolution of metabolism. Here we present the whole-genome metabolic network of this organism and examine in detail the driving forces that have shaped it. We make extensive use of phylometabolic analysis, a method we recently introduced that generates trees of metabolic phenotypes by integrating phylogenetic and metabolic constraints. We reconstruct the evolution of a range of metabolic sub-systems, including the reductive citric acid (rTCA cycle, as well as the biosynthesis and functional roles of several amino acids and cofactors. We show that A. aeolicus uses the reconstructed ancestral pathways within many of these sub-systems, and highlight how the evolutionary interconnections between sub-systems facilitated several key innovations. Our analyses further highlight three general classes of driving forces in metabolic evolution. One is the duplication and divergence of genes for enzymes as these progress from lower to higher substrate specificity, improving the kinetics of certain sub-systems. A second is the kinetic optimization of established pathways through fusion of enzymes, or their organization into larger complexes. The third is the minimization of the ATP unit cost to synthesize biomass, improving thermodynamic efficiency. Quantifying the distribution of these classes of innovations across metabolic sub-systems and across the tree of life will allow us to assess how a tradeoff between maximizing growth rate and growth efficiency has shaped the long-term metabolic evolution of the biosphere.

  6. Phenotypic variation in the plant pathogenic bacterium Acidovorax citrulli.

    Directory of Open Access Journals (Sweden)

    Ram Kumar Shrestha

    Full Text Available Acidovorax citrulli causes bacterial fruit blotch (BFB of cucurbits, a disease that threatens the cucurbit industry worldwide. Despite the economic importance of BFB, little is known about pathogenicity and fitness strategies of the bacterium. We have observed the phenomenon of phenotypic variation in A. citrulli. Here we report the characterization of phenotypic variants (PVs of two strains, M6 and 7a1, isolated from melon and watermelon, respectively. Phenotypic variation was observed following growth in rich medium, as well as upon isolation of bacteria from inoculated plants or exposure to several stresses, including heat, salt and acidic conditions. When grown on nutrient agar, all PV colonies possessed a translucent appearance, in contrast to parental strain colonies that were opaque. After 72 h, PV colonies were bigger than parental colonies, and had a fuzzy appearance relative to parental strain colonies that are relatively smooth. A. citrulli colonies are generally surrounded by haloes detectable by the naked eye. These haloes are formed by type IV pilus (T4P-mediated twitching motility that occurs at the edge of the colony. No twitching haloes could be detected around colonies of both M6 and 7a1 PVs, and microscopy observations confirmed that indeed the PVs did not perform twitching motility. In agreement with these results, transmission electron microscopy revealed that M6 and 7a1 PVs do not produce T4P under tested conditions. PVs also differed from their parental strain in swimming motility and biofilm formation, and interestingly, all assessed variants were less virulent than their corresponding parental strains in seed transmission assays. Slight alterations could be detected in some DNA fingerprinting profiles of 7a1 variants relative to the parental strain, while no differences at all could be seen among M6 variants and parental strain, suggesting that, at least in the latter, phenotypic variation is mediated by slight genetic

  7. Isolation and Determination of Bacterial Soft Rot Pathogens from Potato Tubers in Yunnan%云南马铃薯细菌性软腐病原菌的分离鉴定

    Institute of Scientific and Technical Information of China (English)

    赵志坚; 王淑芬; 方琦; 李先平; 何云昆

    2000-01-01

    从云南省马铃薯产区取样,用厌气技术分离纯化了42个软腐欧文氏杆菌菌株.根据品种差异代表性地选出20个菌株进行主要细菌学性状鉴定,并在此基础上进行分类.结果表明:13个菌株(占65%)是Erwinia carotovora var. carotovora,3个菌株(15%)是Erwinia chrysanthemi,其它4个菌株根据其生理生化特性划归为中间型,介于Erwinia carotovora var. carotovora和Erwinia chrysanthemi之间.

  8. Bioinformatics Analysis on a Novel T3SS Pathogenicity Islands of Erwinia amylovora Ea273 and its Homology in Erwinia%梨火疫病菌新的Ⅲ型泌出系统(T3SS)分析及在其他菌株中的分布

    Institute of Scientific and Technical Information of China (English)

    柴一秋; BOCSANCZY Ana Maria; BEER Steven; 王金生

    2010-01-01

    试验分析了梨火疫病菌(Erwinia amylovora)Ea273基因组上新的T3SS(Trpe Ⅲ Secretion System,T3SS)致病岛特征并验证它们在Erwinia属的21个菌株基因组上是否存在.生物信息学分析发现,Ea273基因组不同位点上存在两个新的T3SS致病岛,称为PAI-2和PAI-3,大小分别为32.889kb和21.792kb.经PCR和Southern blot证明Ea273、Ea246、Ea262、Ea528以及Ea644 5个菌株基因组存在PAI-2和PAI-3.PAI-3和PAI-2在基因结构和组成上相似,同源性超过70%,PAI-3包含PAI-2所有基因,两者和昆虫内生菌S. gtossinidius编码的T3SS的SSR-1致病岛非常相似.

  9. Separation and Identification of Erwinia caratovora ssp . caratovora Pathogen of Coriandrum sativum L%芫荽软腐病病原菌分离与鉴定

    Institute of Scientific and Technical Information of China (English)

    张立微; 张景涛; 李小梅

    2015-01-01

    为有效防治芫荽软腐病,提高其产质量,通过对引起芫荽腐烂而绝产的病害的病原菌进行分离、纯化,对19个菌株进行17项生理生化指标测定。结果表明:根据伯杰氏细菌鉴定手册(第八版)以 Ecc为对照进行鉴定,确定E1(Ames24923)、E3(Ames24927)、E4(CORI107)、E7(CORI139)、E8(CORI140)、E10(CORI147)、E12(CORI289)、E13(CORI318)、E18(PI664510)为芫荽胡萝卜欧文氏杆菌属胡萝卜软腐欧文氏杆菌胡萝卜软腐病亚种(Erw inia carotovora ssp .carotovora ,Ecc)细菌,而其余供试菌株通过测定可确定为细菌,但属何种还需进一步鉴定。%In order to control the Erwinia caratovora ssp .caratovora of Coriandrum sativum L .,and improve the quality ,through the isolating and purificating of the pathogen which causes coriander decay ,19 strains were isolated and purified .The results showed that according to Bergey's Manual of Systematic Bacteriology (eighth edition) and 17 tests of physiology and biochemistry ,9 strainss were Erwiniacarotovora ssp .carotovora ,inclu‐ding E1(Ames24923) ,E3(Ames24927) ,E4(CORI107) ,E7(CORI139) ,E8(CORI140) ,E10(CORI147) ,E12 (CORI289) ,E13(CORI318) ,E18(PI664510) .Other strains should to be identified by other tools .

  10. Characterization of an Endophytic Bacterium G062 Isolate with Beneficial Traits

    Directory of Open Access Journals (Sweden)

    ALINA AKHDIYA

    2014-12-01

    Full Text Available An endophytic bacterium isolate G062 was characterized base on its molecular genetic potents, morphology, physiology, and biochemistry reactions. Analysis of 16S rDNA sequences of G062 showed the highest similarity to Paracoccus halophilus (98%. Detection of the phlD and prnC genes occurrence indicated that the bacterium had this antibiotic-like genes of Diacethylphloroglucinol (DAPG and pyrrolnitrin. The cells are rod shaped (0.59-0.89 x 1.85-3.3 µm, aerobic, Gram negative, non motile, non spore forming, positive catalase, positive oxydase, could reduce NO3 to N2, nitrogen fixing, producing siderophore and plant growth hormones-like compounds (IAA, Gibberellin, and zeatin, and solubilizing phosphate. The G062 isolate could grow on media containing 2.5% NaCl. Range of the temperature and pH growth were 15-40 and 5.0-9.5 oC, respectively. The bacterium did not cause red blood cells lysis. There was no hypersensitive response when it was injected into tobacco leaves, and it was not pathogenic against potato plantlets. Moreover, the bacterium promoted the growth of the potato plant and had high colonization ability. These results suggested that the bacterium had beneficial and good traits as biological agent candidate to promote potato plant growth.

  11. The atherogenic bacterium Porphyromonas gingivalis evades circulating phagocytes by adhering to erythrocytes

    DEFF Research Database (Denmark)

    Belstrøm, Daniel; Holmstrup, Palle; Damgaard, Christian

    2011-01-01

    A relationship between periodontitis and coronary heart disease has been investigated intensively. A pathogenic role for the oral bacterium Porphyromonas gingivalis has been suggested for both diseases. We examined whether complement activation by P. gingivalis strain ATCC 33277 allows the bacter......A relationship between periodontitis and coronary heart disease has been investigated intensively. A pathogenic role for the oral bacterium Porphyromonas gingivalis has been suggested for both diseases. We examined whether complement activation by P. gingivalis strain ATCC 33277 allows...... the bacterium to adhere to human red blood cells (RBCs) and thereby evade attack by circulating phagocytes. On incubation with normal human serum, the P. gingivalis strain efficiently fixed complement component 3 (C3). Incubation of bacteria with washed whole blood cells suspended in autologous serum resulted....... gingivalis exploits RBCs as a transport vehicle, rendering it inaccessible to attack by phagocytes, and by doing so plays a role in the development of systemic diseases....

  12. Action of the Selenomorpholine Compounds on the Bacterium Growth by Microcalorimetry

    Institute of Scientific and Technical Information of China (English)

    李曦; 刘义; 等

    2002-01-01

    The action of β-(N-selenomorpholine) ethyl phenyl ketone hydrochloride and 4-(N-selenomorpholine)-2-butanone hydro-chloride on Escherichia coli and Staphylococcus aureus was studied by microcalorimetry,Differences in their capacities to affect the metabolism of this bacterium were observed.The kinetics shows that the selenomorpholine compounds had action on the metabolism process of Escherichia coli and Staphylococcus aureus.The rate constant (k) of the studied bacterium in the presence of the drugs are concentration-dependant.The growth rate constants decrease with an increase in the mass of the selenomorpholine compounds ,but their relationship is different.As deduced from the rate constant(k) of the studied bacterium(in log phase )and the half inhibitory concentration (IC50),the experimental results reveal that the studied selenomorpholine compounds all have good antibiotic activity and better antibacterial activity on Staphylcoccus aureus than on Escherichia coli.

  13. Action of the Selenomorpholine Compounds on the Bacterium Growth by Microcalorimetry

    Institute of Scientific and Technical Information of China (English)

    LI,Xi(李曦); LIU,Yi(刘义); WU,Jun(吴军); QU,Song-Sheng(屈松生)

    2002-01-01

    The action of β-(N-selenomorpholine) ethyl phenyl ketone hy drochloride and 4-(N-selenomorpholine)-2-butanone hydrochloride on Escherichia coli and Staphylococcus aureus was studied by microcalorimetry. Differences in their capacities to affect the metabolism of this bacterium were observed. The kinetics shows that the selenomorphline compounds had action on the metabolism process of Escherichia coli and Staphylococcus aureus. The rate constant (k) of the studied bacterium in the presence of the drugs are concentration-dependant. The growth rate constants decrease with an increase in the mass of the selenomorpholine compounds, but their relationship is different. As deduced from the rate constant (k) of the studied bacterium (in log phase) and the half inhibitory concentration (IC50), the experimental results reveal that the studied selenomorphline compounds all have good antibiotic activity and better antibacterial activity on Staphylococcus aureus than on Escherichia coli.

  14. Complete Genome Sequence of a thermotolerant sporogenic lactic acid bacterium, Bacillus coagulans strain 36D1

    Energy Technology Data Exchange (ETDEWEB)

    Rhee, Mun Su [University of Florida, Gainesville; Moritz, Brelan E. [University of Florida, Gainesville; Xie, Gary [Los Alamos National Laboratory (LANL); Glavina Del Rio, Tijana [U.S. Department of Energy, Joint Genome Institute; Dalin, Eileen [U.S. Department of Energy, Joint Genome Institute; Tice, Hope [U.S. Department of Energy, Joint Genome Institute; Bruce, David [Los Alamos National Laboratory (LANL); Goodwin, Lynne A. [Los Alamos National Laboratory (LANL); Chertkov, Olga [Los Alamos National Laboratory (LANL); Brettin, Thomas S [ORNL; Han, Cliff [Los Alamos National Laboratory (LANL); Detter, J. Chris [U.S. Department of Energy, Joint Genome Institute; Pitluck, Sam [U.S. Department of Energy, Joint Genome Institute; Land, Miriam L [ORNL; Patel, Milind [University of Florida, Gainesville; Ou, Mark [University of Florida, Gainesville; Harbrucker, Roberta [University of Florida, Gainesville; Ingram, Lonnie O. [University of Florida; Shanmugam, Keelnathan T. [University of Florida

    2011-01-01

    Bacillus coagulans is a ubiquitous soil bacterium that grows at 50-55 C and pH 5.0 and fer- ments various sugars that constitute plant biomass to L (+)-lactic acid. The ability of this spo- rogenic lactic acid bacterium to grow at 50-55 C and pH 5.0 makes this organism an attrac- tive microbial biocatalyst for production of optically pure lactic acid at industrial scale not only from glucose derived from cellulose but also from xylose, a major constituent of hemi- cellulose. This bacterium is also considered as a potential probiotic. Complete genome se- quence of a representative strain, B. coagulans strain 36D1, is presented and discussed.

  15. Complete Genome Sequence of a thermotolerant sporogenic lactic acid bacterium, Bacillus coagulans strain 36D1

    Energy Technology Data Exchange (ETDEWEB)

    Xie, Gary [Los Alamos National Laboratory (LANL); Dalin, Eileen [U.S. Department of Energy, Joint Genome Institute; Tice, Hope [U.S. Department of Energy, Joint Genome Institute; Chertkov, Olga [Los Alamos National Laboratory (LANL); Land, Miriam L [ORNL

    2011-01-01

    Bacillus coagulans is a ubiquitous soil bacterium that grows at 50-55 C and pH 5.0 and fer-ments various sugars that constitute plant biomass to L (+)-lactic acid. The ability of this sporogenic lactic acid bacterium to grow at 50-55 C and pH 5.0 makes this organism an attractive microbial biocatalyst for production of optically pure lactic acid at industrial scale not only from glucose derived from cellulose but also from xylose, a major constituent of hemi-cellulose. This bacterium is also considered as a potential probiotic. Complete genome squence of a representative strain, B. coagulans strain 36D1, is presented and discussed.

  16. Atopobacter phocae gen. nov., sp. nov., a novel bacterium isolated from common seals.

    Science.gov (United States)

    Lawson, P A; Foster, G; Falsen, E; Ohlén, M; Collins, M D

    2000-09-01

    Two strains of a Gram-positive, catalase-negative, facultatively anaerobic, rod-shaped bacterium isolated from common seals were characterized using phenotypic and molecular taxonomic methods. The two strains closely resembled each other based on their biochemical characteristics, and PAGE analysis of whole-cell protein patterns confirmed their close phenotypic affinity. 16S rRNA gene sequencing showed that the two strains were genetically highly related (99.8% sequence similarity) and that they constitute a new line of descent within the lactic acid group of bacteria. The nearest phylogenetic neighbours of the unknown bacterium were Granulicatella spp., with related taxa such as enterococci, carnobacteria, Desemzia incerta, Lactosphaera pasteurii, Melissococcus plutonius, tetragenococci and vagococci more distantly related. Based on phylogenetic and phenotypic evidence it is proposed that the unknown bacterium from seals be classified in a new genus as Atopobacter phocae gen. nov., sp. nov. The type strain of Atopobacter phocae is CCUG 42358T (= CIP 106392T).

  17. Studies on the pathogenic bacterium of ulcer disease in Epinephelus awoara

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

    Studies were conducted to determine the cause of the acute mortality of cage-cultured Epinephelus awoara in the Tong'an Bay of Xiamen, China during the summer of 2002. Predominant bacteria strain TS-628 was isolated from the diseased grouper. The virulence test confirmed that TS-628 was the pathogenic bacterium. Biochemical characteristics of the isolates were determined using the automatic bacterial identification system and standard tube tests. To further confirm the identification, a 1 121 bp 16S rRNA gene sequence of the isolate was amplified by PCR, which had been deposited into Genbank (accession number: AY747308). According to the biochemical characteristics and by comparing the 16S rRNA gene homology of the isolate, the pathogenic bacterium was identified as Vibrio harveyi. Drug sensitivity tests showed that this pathogenic bacterium was sensitive to 16 antibacterials, especially to chloramphenicol and actinospectacin, but completely resistant to antibacterials likes vancomycin, penicillin, lincomycin, and so on.

  18. Phosphate enhances levan production in the endophytic bacterium Gluconacetobacter diazotrophicus Pal5.

    Science.gov (United States)

    Idogawa, Nao; Amamoto, Ryuta; Murata, Kousaku; Kawai, Shigeyuki

    2014-01-01

    Gluconacetobacter diazotrophicus is a gram-negative and endophytic nitrogen-fixing bacterium that has several beneficial effects in host plants; thus, utilization of this bacterium as a biofertilizer in agriculture may be possible. G. diazotrophicus synthesizes levan, a D-fructofuranosyl polymer with β-(2→6) linkages, as an exopolysaccharide and the synthesized levan improves the stress tolerance of the bacterium. In this study, we found that phosphate enhances levan production by G. diazotrophicus Pal5, a wild type strain that showed a stronger mucous phenotype on solid medium containing 28 mM phosphate than on solid medium containing 7 mM phosphate. A G. diazotrophicus Pal5 levansucrase disruptant showed only a weak mucous phenotype regardless of the phosphate concentration, indicating that the mucous phenotype observed on 28 mM phosphate medium was caused by levan. To our knowledge, this is the first report of the effect of a high concentration of phosphate on exopolysaccharide production.

  19. Expression of the Bacillus thuringiensis mosquitocidal toxin Cry11Aa in the aquatic bacterium Asticcacaulis excentricus.

    Science.gov (United States)

    Armengol, Gemma; Guevara, Oscar Enrique; Orduz, Sergio; Crickmore, Neil

    2005-12-01

    A mosquitocidal aquatic bacterium has been developed by introducing an operon containing the cry11Aa, and p20 genes from Bacillus thuringiensis subsp. israelensis (Bti) into the gram-negative aquatic bacterium Asticcacaulis excentricus. After transformation, the cry11Aa gene was successfully expressed in recombinant A. excentricus under the tac promoter, at the level of 0.04 pg/cell. The recombinant bacteria were toxic to Aedes aegypti larvae with an LC(50) of 6.83 x 10(5) cells/mL. We believe that these bacteria may have potential as genetically engineered microorganisms for the control of mosquito larvae.

  20. Isolation and characterization of Caldicellulosiruptor lactoaceticus sp. nov., an extremely thermophilic, cellulolytic, anaerobic bacterium

    DEFF Research Database (Denmark)

    Mladenovska, Zuzana; Mathrani, Indra M.; Ahring, Birgitte Kiær

    1995-01-01

    activity. The G + C content of the cellular DNA of strain 6A was 35.2 +/- 0.8 mol%. Complete 16S rDNA sequence analysis showed that strain 6A was phylogenetically related to Caldicellulosiruptor saccharolyticus. It is proposed that the isolated bacterium be named Caldicellulosiruptor lactoaceticus sp. nov....... and ethanol occurred as minor fermentation products. Only a restricted number of carbon sources (cellulose, xylan, starch, pectin, cellobiose, xylose, maltose and lactose) were used as substrates. During growth on Avicel, the bacterium produced free cellulases with carboxymethylcellulase and avicelase...

  1. Homology-based modeling of the Erwinia amylovora type III secretion chaperone DspF used to identify amino acids required for virulence and interaction with the effector DspE.

    Science.gov (United States)

    Triplett, Lindsay R; Wedemeyer, William J; Sundin, George W

    2010-09-01

    The structure of DspF, a type III secretion system (T3SS) chaperone required for virulence of the fruit tree pathogen Erwinia amylovora, was modeled based on predicted structural homology to characterized T3SS chaperones. This model guided the selection of 11 amino acid residues that were individually mutated to alanine via site-directed mutagenesis. Each mutant was assessed for its effect on virulence complementation, dimerization and interaction with the N-terminal chaperone-binding site of DspE. Four amino acid residues were identified that did not complement the virulence defect of a dspF knockout mutant, and three of these residues were required for interaction with the N-terminus of DspE. This study supports the significance of the predicted beta-sheet helix-binding groove in DspF chaperone function.

  2. Toxicity Measurement of Several Bactericides on Erwinia carotovora susp. Atroseptica%几种杀细菌剂对胡萝卜软腐欧文氏菌的毒力测定

    Institute of Scientific and Technical Information of China (English)

    雷玉明; 张建朝; 邢会琴; 费永祥

    2010-01-01

    采用室内抑菌圈法测定了6种杀细菌剂对胡萝卜软腐欧文氏菌马铃薯黑胫病亚种(Erwinia carotovora susp. Atroseptica)的毒力,筛选出2种有明显抑菌作用的药剂72%农用硫酸链霉素可湿性粉剂和90%链霉素·土可溶性粉剂,抑菌效果显著,EC50分别为0.092 1 mg·mL-1和0.097 2 mg·mL-1.

  3. 欧文氏菌乳糖酶的分离纯化及其酶学性质研究%Purification and enzymatic characterization of a β-galactosidase from Erwinia sp.

    Institute of Scientific and Technical Information of China (English)

    夏雨; 成玉梁; 赵莹; 吕源玲; 孙震

    2011-01-01

    The β-galactosidase from Erwinia sp. E5 was isolated and purified. The specific enzymatic activity of the purified enzyme was 554. 92 U/mg protein. The properties of this enzyme were characterized. Results showed this enzyme had a maximum activity at 40 ℃ , and more than 40% of the enzymatic activity was kept at 20 ℃. The optimal pH to this enzyme is 7. 0, and 95% of the enzymatic activity was kept under pH 6. 5. Metal cations Mg2+ and Na+ could activate the enzymatic activity while the cation Ca2+ acted as a weak inhibitor. The lactose hydrolysis experiment showed that this enzyme kept a relatively fast reaction speed when the lactose concentration was below 10%.%对欧文氏菌Erwinia sp.E5株所产β-半乳糖苷酶进行分离纯化,所得纯酶的比酶活为554.92 U/mg ·蛋白质.对纯化得到的酶进行酶学性质研究,其最适反应温度为40℃,在20℃保持40%以上的酶活力;该酶最适反应pH为7.0,在pH6.5体系中保持95%的酶活力;Mg2+和Na+对该酶具有激活作用,而Ca2+为弱抑制剂.乳糖水解试验显示该酶在乳糖含量小于10%的反应体系中具有较高反应速率.

  4. Draft Genome Sequence of an Anaerobic and Extremophilic Bacterium, Caldanaerobacter yonseiensis, Isolated from a Geothermal Hot Stream

    OpenAIRE

    2013-01-01

    Caldanaerobacter yonseiensis is a strictly anaerobic, thermophilic, spore-forming bacterium, which was isolated from a geothermal hot stream in Indonesia. This bacterium utilizes xylose and produces a variety of proteases. Here, we report the draft genome sequence of C. yonseiensis, which reveals insights into the pentose phosphate pathway and protein degradation metabolism in thermophilic microorganisms.

  5. Isolation from the Sorghum bicolor Mycorrhizosphere of a Bacterium Compatible with Arbuscular Mycorrhiza Development and Antagonistic towards Soilborne Fungal Pathogens

    Science.gov (United States)

    Budi, S. W.; van Tuinen, D.; Martinotti, G.; Gianinazzi, S.

    1999-01-01

    A gram-positive bacterium with antagonistic activity towards soilborne fungal pathogens has been isolated from the mycorrhizosphere of Sorghum bicolor inoculated with Glomus mosseae. It has been identified as Paenibacillus sp. strain B2 based on its analytical profile index and on 16S ribosomal DNA analysis. Besides having antagonistic activity, this bacterium stimulates mycorrhization. PMID:10543835

  6. Draft Genome Sequence of Bacillus licheniformis Strain GB2, a Hydrocarbon-Degrading and Plant Growth-Promoting Soil Bacterium.

    Science.gov (United States)

    Gkorezis, Panagiotis; Van Hamme, Jonathan; Bottos, Eric; Thijs, Sofie; Balseiro-Romero, Maria; Monterroso, Carmela; Kidd, Petra Suzan; Rineau, Francois; Weyens, Nele; Sillen, Wouter; Vangronsveld, Jaco

    2016-06-23

    We report the 4.39 Mb draft genome of Bacillus licheniformis GB2, a hydrocarbonoclastic Gram-positive bacterium of the family Bacillaceae, isolated from diesel-contaminated soil at the Ford Motor Company site in Genk, Belgium. Strain GB2 is an effective plant-growth promoter useful for diesel fuel remediation applications based on plant-bacterium associations.

  7. Identification and functional analysis of a phytoene desaturase gene from the extremely radioresistant bacterium Deinococcus radiodurans.

    Science.gov (United States)

    Xu, Zhenjian; Tian, Bing; Sun, Zongtao; Lin, Jun; Hua, Yuejin

    2007-05-01

    The phytoene-related desaturases are the key enzymes in the carotenoid biosynthetic pathway. The gene encoding phytoene desaturase in the deinoxanthin synthesis pathway of Deinococcus radiodurans was identified and characterized. Two putative phytoene desaturase homologues (DR0861 and DR0810) were identified by analysis of conserved amino acid regions, and the former displayed the highest identity (68 %) with phytoene desaturase of the cyanobacterium Gloeobacter violaceus. DR0861 gene knockout and dinucleotide-binding motif deletion resulted in the arrest of lycopene synthesis and the accumulation of phytoene. The colourless DR0861 knockout mutant became more sensitive to acute ionizing radiation and oxygen stress. Complementation of the mutant with a heterologous or homologous gene restored its pigment and resistance. The desaturase activity of DR0861 (crtI) was further confirmed by the assay of enzyme activity in vitro and heterologous expression in Escherichia coli containing crtE and crtB genes (responsible for phytoene synthesis) from Erwinia uredovora. In addition, the amount of lycopene synthesis in E. coli resulting from the expression of crtI from D. radiodurans was determined, and this had significant dose-dependent effects on the survival rate of E. coli exposed to hydrogen peroxide and ionizing radiation.

  8. Complete genome sequence of the xylan-degrading subseafloor bacterium Microcella alkaliphila JAM-AC0309.

    Science.gov (United States)

    Kurata, Atsushi; Hirose, Yuu; Misawa, Naomi; Wakazuki, Sachiko; Kishimoto, Noriaki; Kobayashi, Tohru

    2016-03-10

    Here we report the complete genome sequence of Microcella alkaliphila JAM-AC0309, which was newly isolated from the deep subseafloor core sediment from offshore of the Shimokita Peninsula of Japan. An array of genes related to utilization of xylan in this bacterium was identified by whole genome analysis.

  9. Comment on "A bacterium that degrades and assimilates poly(ethylene terephthalate)".

    Science.gov (United States)

    Yang, Yu; Yang, Jun; Jiang, Lei

    2016-08-19

    Yoshida et al (Report, 11 March 2016, p. 1196) reported that the bacterium Ideonella sakaiensis 201-F6 can degrade and assimilate poly(ethylene terephthalate) (PET). However, the authors exaggerated degradation efficiency using a low-crystallinity PET and presented no straightforward experiments to verify depolymerization and assimilation of PET. Thus, the authors' conclusions are rather misleading.

  10. Isolation and algae-lysing characteristics of the algicidal bacterium B5

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    Water blooms have become a worldwide environmental problem. Recently, algicidal bacteria have attracted wide attention as possible agents for inhibiting algal water blooms. In this study, one strain of algicidal bacterium B5 was isolated from activated sludge. On the basis of analysis of its physiological characteristics and 16S rDNA gene sequence, it was identified as Bacillus fusiformis. Its algae-lysing characteristics on Microcystis aeruginosa, Chlorella and Scenedesmus were tested. The results showed that: (1) the algicidal bacterium B5 is a Gram-negative bacterium. The 16S rDNA nucleotide sequence homology of strain B5 with 2 strains of B. fusiformis reached 99.86%, so B5 was identified as B. fusiformis; (2) the algal-lysing effects of the algicidal bacterium B5 on M. aeruginosa, Chlorella and Scenedesmus were pronounced. The initial bacterial and algal cell densities strongly influence the removal rates of chlorophyll-a. The greater the initial bacterial cell density, the faster the degradation of chlorophyll-a. The greater the initial algal cell density, the slower the degradation of chlorophyll-a. When the bacterial cell density was 3.6 × 107 cells/ml, nearly 90% of chlorophyll-a was removed. When the chlorophyll-a concentration was less than 550 μg/L, about 70 % was removed; (3) the strain B5 lysed algae not directly but by secreting metabolites and these metabolites could bear heat treatment.

  11. Inactivation of Glutamine Synthetase by Ammonia Shock in the Gram-Positive Bacterium Streptomyces cattleya

    OpenAIRE

    Wax, Richard; Synder, Linda; Kaplan, Louis

    1982-01-01

    In cultures of the gram-positive bacterium Streptomyces cattleya, a rapid inactivation of glutamine synthetase was seen after ammonia shock. pH activity curves for ammonia-shocked and control cultures are shown. A peak of glutamine synthetase activity was seen during fermentation for production of the antibiotic thienamycin.

  12. Draft Genome Sequence of Desulfuromonas acetexigens Strain 2873, a Novel Anode-Respiring Bacterium

    KAUST Repository

    Katuri, Krishna

    2017-03-03

    Here, we report the draft genome sequence of Desulfuromonas acetexigens strain 2873, which was originally isolated from digester sludge from a sewage treatment plant in Germany. This bacterium is capable of anode respiration with high electrochemical activity in microbial electrochemical systems. The draft genome contains 3,376 predicted protein-coding genes and putative multiheme c-type cytochromes.

  13. Modeling of Cd Uptake and Efflux Kinetics in Metal-Resistant Bacterium Cupriavidus metallidurans

    NARCIS (Netherlands)

    Hajdu, R.; Pinheiro, J.P.; Galceran, J.; Slaveykova, V.I.

    2010-01-01

    The Model of Uptake with Instantaneous Adsorption and Efflux, MUIAE, describing and predicting the overall Cd uptake by the metal-resistant bacterium Cupriavidus metallidurans CH34, is presented. MUIAE takes into account different processes at the bacteria-medium interface with specific emphasis on

  14. Photobacterium marinum sp. nov., a marine bacterium isolated from a sediment sample from Palk Bay, India

    Digital Repository Service at National Institute of Oceanography (India)

    Srinivas, T.N.R.; VijayaBhaskar, Y.; Bhumika, V.; AnilKumar, P.

    histaminum sp. nov., a histamine-producing marine bacterium. Int. J. Syst. Bacteriol. 44, 631-636. [20] Ostle, A.G., Holt, J.G. (1982) Nile blue A as fluorescent stain for poly-b-hydroxybutyrate. Appl. Environ. Microbiol. 44, 238-241. [21] Park, Y...

  15. Fluoroacetate biosynthesis from the marine-derived bacterium Streptomyces xinghaiensis NRRL B-24674.

    Science.gov (United States)

    Huang, Sheng; Ma, Long; Tong, Ming Him; Yu, Yi; O'Hagan, David; Deng, Hai

    2014-07-21

    Genome sequencing identified a fluorinase gene in the marine bacterium Streptomyces xinghaiensis NRRL B-24674. Fermentation of the organism with inorganic fluoride (2 mM) demonstrated that the organism could biosynthesise fluoroacetate and that fluoroacetate production is sea-salt dependent. This is the first fluorometabolite producing microorganism identified from the marine environment.

  16. Biohydrogen Production by the Thermophilic Bacterium Caldicellulosiruptor saccharolyticus: Current Status and Perspectives

    NARCIS (Netherlands)

    Bielen, A.A.M.; Verhaart, M.R.A.; Oost, van der J.; Kengen, S.W.M.

    2013-01-01

    Caldicellulosiruptor saccharolyticus is one of the most thermophilic cellulolytic organisms known to date. This Gram-positive anaerobic bacterium ferments a broad spectrum of mono-, di- and polysaccharides to mainly acetate, CO2 and hydrogen. With hydrogen yields approaching the theoretical limit fo

  17. Genome sequence of the mycorrhizal helper bacterium Pseudomonas fluorescens BBc6R8

    Energy Technology Data Exchange (ETDEWEB)

    Deveau, Aurelie [French National Insitute for Agricultural Research (INRA); Grob, Harald [University of Bonn, Germany; Morin, Emmanuelle [INRA, Nancy, France; Karpinets, Tatiana V [ORNL; Utturkar, Sagar M [ORNL; Mehnaz, Samina [University of the Punjab, Pakistan; Kurz, Sven [University of Bonn, Germany; Martin, Francis [INRA, Nancy, France; Frey-Klett, Pascale [INRA, Nancy, France; Labbe, Jessy L [ORNL

    2014-01-01

    We report the draft genome sequence of the mycorrhiza helper bacterium Pseudomonas fluorescens strain BBc6R8 . Several traits which could be involved in the mycorrhiza helper ability of the bacterial strain such as multiple secretion systems, auxin metabolism and phosphate mobilization were evidenced in the genome.

  18. Robinsoniella peoriensis: A model anaerobic commensal bacterium for acquisition of antibiotic resistance?

    Science.gov (United States)

    Background: R. peoriensis was characterized in our laboratories from swine manure and feces as a Gram-positive, anaerobic bacterium. Since then strains of this species have been identified from a variety of mammalian and other gastrointestinal (GI) tracts, suggesting it is a member of the commensal ...

  19. Flavobacterium nitratireducens sp. nov., an amylolytic bacterium of the family Flavobacteriaceae isolated from coastal surface seawater

    Digital Repository Service at National Institute of Oceanography (India)

    Nupur; Bhumika, V.; Srinivas, T.N.R.; AnilKumar, P.

    A novel Gram-negative, rod-shaped, non-motile bacterium, designated strain N1 sup(T), was isolated from a marine water sample collected from the sea shore, Bay of Bengal, Visakhapatnam, India. The strain was positive for starch hydrolysis, nitrate...

  20. Marinilabilia nitratireducens sp. nov., a lipolytic bacterium of the family Marinilabiliaceae isolated from marine solar saltern

    Digital Repository Service at National Institute of Oceanography (India)

    Shalley, S.; PradipKumar; Srinivas, T.N.R.; Suresh, K.; AnilKumar, P.

    A Gram-negative, rod shaped, motile bacterium, was isolated from a marine solar saltern sample collected from Kakinada, India. Strain AK2 sup(T) was determined to be positive for nitrate reduction, catalase, Ala-Phe-Pro-arylamidase, beta...

  1. Draft Genome Sequence of the Moderately Thermophilic Bacterium Schleiferia thermophila Strain Yellowstone (Bacteroidetes).

    Science.gov (United States)

    Thiel, Vera; Hamilton, Trinity L; Tomsho, Lynn P; Burhans, Richard; Gay, Scott E; Ramaley, Robert F; Schuster, Stephan C; Steinke, Laurey; Bryant, Donald A

    2014-08-28

    The draft genome sequence of the moderately thermophilic bacterium Schleiferia thermophila strain Yellowstone (Bacteroidetes), isolated from Octopus Spring (Yellowstone National Park, WY, USA) was sequenced and comprises 2,617,694 bp in 35 contigs. The draft genome is predicted to encode 2,457 protein coding genes and 37 tRNA encoding genes and two rRNA operons.

  2. Transcriptome analysis of the rhizosphere bacterium Azospirillum brasilense reveals an extensive auxin response.

    Science.gov (United States)

    Van Puyvelde, Sandra; Cloots, Lore; Engelen, Kristof; Das, Frederik; Marchal, Kathleen; Vanderleyden, Jos; Spaepen, Stijn

    2011-05-01

    The rhizosphere bacterium Azospirillum brasilense produces the auxin indole-3-acetic acid (IAA) through the indole-3-pyruvate pathway. As we previously demonstrated that transcription of the indole-3-pyruvate decarboxylase (ipdC) gene is positively regulated by IAA, produced by A. brasilense itself or added exogenously, we performed a microarray analysis to study the overall effects of IAA on the transcriptome of A. brasilense. The transcriptomes of A. brasilense wild-type and the ipdC knockout mutant, both cultured in the absence and presence of exogenously added IAA, were compared.Interfering with the IAA biosynthesis/homeostasis in A. brasilense through inactivation of the ipdC gene or IAA addition results in much broader transcriptional changes than anticipated. Based on the multitude of changes observed by comparing the different transcriptomes, we can conclude that IAA is a signaling molecule in A. brasilense. It appears that the bacterium, when exposed to IAA, adapts itself to the plant rhizosphere, by changing its arsenal of transport proteins and cell surface proteins. A striking example of adaptation to IAA exposure, as happens in the rhizosphere, is the upregulation of a type VI secretion system (T6SS) in the presence of IAA. The T6SS is described as specifically involved in bacterium-eukaryotic host interactions. Additionally, many transcription factors show an altered regulation as well, indicating that the regulatory machinery of the bacterium is changing.

  3. Draft Genome Sequence of a Thermophilic Desulfurization Bacterium, Geobacillus thermoglucosidasius Strain W-2

    Science.gov (United States)

    Zhu, Lin; Li, Mingchang; Guo, Shuyi

    2016-01-01

    Geobacillus thermoglucosidasius strain W-2 is a thermophilic bacterium isolated from a deep-subsurface oil reservoir in northern China, which is capable of degrading organosulfur compounds. Here, we report the draft genome sequence of G. thermoglucosidasius strain W-2, which may help to elucidate the genetic basis of biodegradation of organosulfur pollutants under heated conditions. PMID:27491977

  4. Genome Sequence of the Acetogenic Bacterium Moorella mulderi DSM 14980T

    Science.gov (United States)

    Castillo Villamizar, Genis Andrés

    2016-01-01

    Here, we report the draft genome sequence of Moorella mulderi DSM 14980T, a thermophilic acetogenic bacterium, which is able to grow autotrophically on H2 plus CO2 using the Wood-Ljungdahl pathway. The genome consists of a circular chromosome (2.99 Mb). PMID:27231372

  5. Bacterium induces cryptic meroterpenoid pathway in the pathogenic fungus Aspergillus fumigatus.

    Science.gov (United States)

    König, Claudia C; Scherlach, Kirstin; Schroeckh, Volker; Horn, Fabian; Nietzsche, Sandor; Brakhage, Axel A; Hertweck, Christian

    2013-05-27

    Stimulating encounter: The intimate, physical interaction between the soil-derived bacterium Streptomyces rapamycinicus and the human pathogenic fungus Aspergillus fumigatus led to the activation of an otherwise silent polyketide synthase (PKS) gene cluster coding for an unusual prenylated polyphenol (fumicycline A). The meroterpenoid pathway is regulated by a pathway-specific activator gene as well as by epigenetic factors.

  6. Moritella viscosa, a pathogenic bacterium affecting the fillet quality in fish

    DEFF Research Database (Denmark)

    Ingerslev, Hans-Christian; Nielsen, Michael Engelbrecht

    2011-01-01

    ’ which affects various fish species in seawater during cold periods (Lunder et al. 1995). The bacterium is mainly a problem for farmed salmonid species, such as Atlantic salmon (Salmo salar) and rainbow trout (Oncorhynchus mykiss), but has also been isolated from other fish species, including Atlantic...

  7. Genome sequence of Citrobacter sp. strain A1, a dye-degrading bacterium.

    Science.gov (United States)

    Chan, Giek Far; Gan, Han Ming; Rashid, Noor Aini Abdul

    2012-10-01

    Citrobacter sp. strain A1, isolated from a sewage oxidation pond, is a facultative aerobe and mesophilic dye-degrading bacterium. This organism degrades azo dyes efficiently via azo reduction and desulfonation, followed by the successive biotransformation of dye intermediates under an aerobic environment. Here we report the draft genome sequence of Citrobacter sp. A1.

  8. The construction of an engineered bacterium to remove cadmium from wastewater.

    Science.gov (United States)

    Chang, S; Shu, H

    2014-01-01

    The removal of cadmium (Cd) from wastewater before it is released from factories is important for protecting human health. Although some researchers have developed engineered bacteria, the resistance of these engineered bacteria to Cd have not been improved. In this study, two key genes involved in glutathione synthesis (gshA and gshB), a serine acetyltransferase gene (cysE), a Thlaspi caerulescens phytochelatin synthase gene (TcPCS1), and a heavy metal ATPase gene (TcHMA3) were transformed into Escherichia coli BL21. The resistance of the engineered bacterium to Cd was significantly greater than that of the initial bacterium and the Cd accumulation in the engineered bacterium was much higher than in the initial bacterium. In addition, the Cd resistance of the bacteria harboring gshB, gshA, cysE, and TcPCS1 was higher than that of the bacteria harboring gshA, cysE, and TcPCS1. This finding demonstrated that gshB played an important role in glutathione synthesis and that the reaction catalyzed by glutathione synthase was the limiting step for producing phytochelatins. Furthermore, TcPCS1 had a greater specificity and a higher capacity for removing Cd than SpPCS1, and TcHMA3 not only played a role in T. caerulescens but also functioned in E. coli.

  9. Complete genome of Nitrosospira briensis C-128, an ammonia-oxidizing bacterium from agricultural soil

    NARCIS (Netherlands)

    Rice, Marlen C.; Norton, Jeanette M.; Valois, Frederica; Bollmann, Annette; Bottomley, Peter J.; Klotz, Martin G.; Laanbroek, Hendrikus J.; Suwa, Yuichi; Stein, Lisa Y.; Sayavedra-Soto, Luis; Woyke, Tanja; Shapiro, Nicole; Goodwin, Lynne A.; Huntemann, Marcel; Clum, Alicia; Pillay, Manoj; Kyrpides, Nikos; Varghese, Neha; Mikhailova, Natalia; Markowitz, Victor; Palaniappan, Krishna; Ivanova, Natalia; Stamatis, Dimitrios; Reddy, T. B. K.; Ngan, Chew Yee; Daum, Chris

    2016-01-01

    Nitrosospira briensis C-128 is an ammonia-oxidizing bacterium isolated from an acid agricultural soil. N. briensis C-128 was sequenced with PacBio RS technologies at the DOE-Joint Genome Institute through their Community Science Program (2010). The high-quality finished genome contains one chromosom

  10. Marinobacter nitratireducens sp. nov., a halophilic and lipolytic bacterium isolated from coastal surface sea water

    Digital Repository Service at National Institute of Oceanography (India)

    Bhumika, V.; Ravinder, K.; Korpole, S.; Srinivas, T.N.R.; AnilKumar, P.

    A novel Gram-stain-negative, rod-shaped, motile bacterium, designated strain AK21T , was isolated from coastal surface sea water at Visakhapatnam, India. The strain was positive for oxidase, catalase, lipase, L-proline arylamidase...

  11. Draft Genome Sequence of Photorhabdus luminescens subsp. laumondii HP88, an Entomopathogenic Bacterium Isolated from Nematodes

    OpenAIRE

    Ghazal, Shimaa; Oshone, Rediet; Simpson, Stephen,; Morris, Krystalynne; Abebe-Akele, Feseha; Thomas, W. Kelley; Khalil, Kamal M.; Tisa, Louis S.

    2016-01-01

    Photorhabdus luminescens subsp. laumondii HP88 is an entomopathogenic bacterium that forms a symbiotic association with Heterorhabditis nematodes. We report here a 5.27-Mbp draft genome sequence for P. luminescens subsp. laumondii HP88, with a G+C content of 42.4% and containing 4,243 candidate protein-coding genes.

  12. Purification and reconstitution of the glutamate carrier GltT of the thermophilic bacterium Bacillus stearothermophilus

    NARCIS (Netherlands)

    Gaillard, Isabelle; Slotboom, Dirk-Jan; Knol, Jan; Lolkema, Juke S.; Konings, Wil N.

    1996-01-01

    An affinity tag consisting of six adjacent histidine residues followed by an enterokinase cleavage site was genetically engineered at the N-terminus of the glutamate transport protein GltT of the thermophilic bacterium Bacillus stearothermophilus. The fusion protein was expressed in Escherichia coli

  13. Active efflux systems in the solvent-tolerant bacterium Pseudomonas putida S12

    NARCIS (Netherlands)

    Kieboom, J.

    2002-01-01

    The aim of the research presented in this thesis was to study the molecular mechanisms of organic solvent tolerance in Pseudomonas putida S12. This bacterium is capable of growth at saturated solvent concentrations, which are lethal to normal bacteria. Organic solve

  14. Draft Genome Sequence of Burkholderia cenocepacia Strain 869T2, a Plant-Beneficial Endophytic Bacterium.

    Science.gov (United States)

    Ho, Ying-Ning; Huang, Chieh-Chen

    2015-11-12

    An endophytic bacterium, Burkholderia cenocepacia 869T2, isolated from vetiver grass, has shown its abilities for both in planta biocontrol and plant growth promotion. Its draft genome sequence was determined to provide insights into those metabolic pathways involved in plant-beneficial activity. This is the first genome report for endophytic B. cenocepacia.

  15. Hydrogen Production by Co-cultures of Rhizopus oryzae and a Photosynthetic Bacterium, Rhodobacter sphaeroides RV

    Science.gov (United States)

    Asada, Yasuo; Ishimi, Katsuhiro; Nagata, Yoko; Wakayama, Tatsuki; Miyake, Jun; Kohno, Hideki

    Hydrogen production with glucose by using co-immobilized cultures of a fungus, Rhizopus oryzae NBRC5384, and a photosynthetic bacterium, Rhodobacter sphaeroides RV, in agar gels was studied. The co-immobilized cultures converted glucose to hydrogen via lactate in a high molar yield of about 8moles of hydrogen per glucose at a maximum under illuminated conditions.

  16. Inactivation of Glutamine Synthetase by Ammonia Shock in the Gram-Positive Bacterium Streptomyces cattleya.

    Science.gov (United States)

    Wax, R; Synder, L; Kaplan, L

    1982-10-01

    In cultures of the gram-positive bacterium Streptomyces cattleya, a rapid inactivation of glutamine synthetase was seen after ammonia shock. pH activity curves for ammonia-shocked and control cultures are shown. A peak of glutamine synthetase activity was seen during fermentation for production of the antibiotic thienamycin.

  17. Draft Genome Sequence of the Moderately Halophilic Bacterium Pseudoalteromonas ruthenica Strain CP76.

    Science.gov (United States)

    de la Haba, Rafael R; Sánchez-Porro, Cristina; León, María José; Papke, R Thane; Ventosa, Antonio

    2013-05-23

    Pseudoalteromonas ruthenica strain CP76, isolated from a saltern in Spain, is a moderately halophilic bacterium belonging to the Gammaproteobacteria. Here we report the draft genome sequence, which consists of a 4.0-Mb chromosome, of this strain, which is able to produce the extracellular enzyme haloprotease CPI.

  18. Engineering a predatory bacterium as a proficient killer agent for intracellular bio-products recovery

    DEFF Research Database (Denmark)

    Martinez, Virginia; Herencias, Cristina; Jurkevitch, Edouard;

    2016-01-01

    This work examines the potential of the predatory bacterium Bdellovibrio bacteriovorus HD100, an obligate predator of other Gram-negative bacteria, as an external cell-lytic agent for recovering valuable intracellular bio-products produced by prey cultures. The bio-product targets to be recovered...

  19. A commensal symbiotic interrelationship for the growth of Symbiobacterium toebii with its partner bacterium, Geobacillus toebii

    Directory of Open Access Journals (Sweden)

    Masui Ryoji

    2011-10-01

    Full Text Available Abstract Background Symbiobacterium toebii is a commensal symbiotic thermophile that absolutely requires its partner bacterium Geobacillus toebii for growth. Despite development of an independent cultivation method using cell-free extracts, the growth of Symbiobacterium remains unknown due to our poor understanding of the symbiotic relationship with its partner bacterium. Here, we investigated the interrelationship between these two bacteria for growth of S. toebii using different cell-free extracts of G. toebii. Results Symbiobacterium toebii growth-supporting factors were constitutively produced through almost all growth phases and under different oxygen tensions in G. toebii, indicating that the factor may be essential components for growth of G. toebii as well as S. toebii. The growing conditions of G. toebii under different oxygen tension dramatically affected to the initial growth of S. toebii and the retarded lag phase was completely shortened by reducing agent, L-cysteine indicating an evidence of commensal interaction of microaerobic and anaerobic bacterium S. toebii with a facultative aerobic bacterium G. toebii. In addition, the growth curve of S. toebii showed a dependency on the protein concentration of cell-free extracts of G. toebii, demonstrating that the G. toebii-derived factors have nutrient-like characters but not quorum-sensing characters. Conclusions Not only the consistent existence of the factor in G. toebii during all growth stages and under different oxygen tensions but also the concentration dependency of the factor for proliferation and optimal growth of S. toebii, suggests that an important biosynthetic machinery lacks in S. toebii during evolution. The commensal symbiotic bacterium, S. toebii uptakes certain ubiquitous and essential compound for its growth from environment or neighboring bacteria that shares the equivalent compounds. Moreover, G. toebii grown under aerobic condition shortened the lag phase of S

  20. Draft genome of an Aerophobetes bacterium reveals a facultative lifestyle in deep-sea anaerobic sediments

    KAUST Repository

    Wang, Yong

    2016-07-01

    Aerophobetes (or CD12) is a recently defined bacterial phylum, of which the metabolic processes and ecological importance remain unclear. In the present study, we obtained the draft genome of an Aerophobetes bacterium TCS1 from saline sediment near the Thuwal cold seep in the Red Sea using a genome binning method. Analysis of 16S rRNA genes of TCS1 and close relatives revealed wide distribution of Aerophobetes in deep-sea sediments. Phylogenetic relationships showed affinity between Aerophobetes TCS1 and some thermophilic bacterial phyla. The genome of TCS1 (at least 1.27 Mbp) contains a full set of genes encoding core metabolic pathways, including glycolysis and pyruvate fermentation to produce acetyl-CoA and acetate. The identification of cross-membrane sugar transporter genes further indicates its potential ability to consume carbohydrates preserved in the sediment under the microbial mat. Aerophobetes bacterium TCS1 therefore probably carried out saccharolytic and fermentative metabolism. The genes responsible for autotrophic synthesis of acetyl-CoA via the Wood–Ljungdahl pathway were also found in the genome. Phylogenetic study of the essential genes for the Wood–Ljungdahl pathway implied relative independence of Aerophobetes bacterium from the known acetogens and methanogens. Compared with genomes of acetogenic bacteria, Aerophobetes bacterium TCS1 genome lacks the genes involved in nitrogen metabolism, sulfur metabolism, signal transduction and cell motility. The metabolic activities of TCS1 might depend on geochemical conditions such as supplies of CO2, hydrogen and sugars, and therefore the TCS1 might be a facultative bacterium in anaerobic saline sediments near cold seeps. © 2016, Science China Press and Springer-Verlag Berlin Heidelberg.

  1. A fragile metabolic network adapted for cooperation in the symbiotic bacterium Buchnera aphidicola

    Directory of Open Access Journals (Sweden)

    Goryanin Igor

    2009-02-01

    Full Text Available Abstract Background In silico analyses provide valuable insight into the biology of obligately intracellular pathogens and symbionts with small genomes. There is a particular opportunity to apply systems-level tools developed for the model bacterium Escherichia coli to study the evolution and function of symbiotic bacteria which are metabolically specialised to overproduce specific nutrients for their host and, remarkably, have a gene complement that is a subset of the E. coli genome. Results We have reconstructed and analysed the metabolic network of the γ-proteobacterium Buchnera aphidicola (symbiont of the pea aphid as a model for using systems-level approaches to discover key traits of symbionts with small genomes. The metabolic network is extremely fragile with > 90% of the reactions essential for viability in silico; and it is structured so that the bacterium cannot grow without producing the essential amino acid, histidine, which is released to the insect host. Further, the amount of essential amino acid produced by the bacterium in silico can be controlled by host supply of carbon and nitrogen substrates. Conclusion This systems-level analysis predicts that the fragility of the bacterial metabolic network renders the symbiotic bacterium intolerant of drastic environmental fluctuations, whilst the coupling of histidine production to growth prevents the bacterium from exploiting host nutrients without reciprocating. These metabolic traits underpin the sustained nutritional contribution of B. aphidicola to the host and, together with the impact of host-derived substrates on the profile of nutrients released from the bacteria, point to a dominant role of the host in controlling the symbiosis.

  2. Draft genome of an Aerophobetes bacterium reveals a facultative lifestyle in deep-sea anaerobic sediments

    Institute of Scientific and Technical Information of China (English)

    Yong Wang; Zhao-Ming Gao; Jiang-Tao Li; Salim Bougouffa; Ren Mao Tian; Vladimir B.Bajic; Pei-Yuan Qian

    2016-01-01

    Aerophobetes (or CD12) is a recently defined bacterial phylum,of which the metabolic processes and ecological importance remain unclear.In the present study,we obtained the draft genome of an Aerophobetes bacterium TCS1 from saline sediment near the Thuwal cold seep in the Red Sea using a genome binning method.Analysis of 16S rRNA genes of TCS1 and close relatives revealed wide distribution of Aerophobetes in deep-sea sediments.Phylogenetic relationships showed affinity between Aerophobetes TCS1 and some thermophilic bacterial phyla.The genome of TCS1 (at least 1.27 Mbp)contains a full set of genes encoding core metabolic pathways,including glycolysis and pyruvate fermentation to produce acetyl-CoA and acetate.The identification of cross-membrane sugar transporter genes further indicates its potential ability to consume carbohydrates preserved in the sediment under the microbial mat.Aerophobetes bacterium TCS1 therefore probably carried out saccharolytic and fermentative metabolism.The genes responsible for autotrophic synthesis of acetyl-CoA via the Wood-Ljungdahl pathway were also found in the genome.Phylogenetic study of the essential genes for the Wood-Ljungdahl pathway implied relative independence of Aerophobetes bacterium from the known acetogens and methanogens.Compared with genomes of acetogenic bacteria,Aerophobetes bacterium TCS 1 genome lacks the genes involved in nitrogen metabolism,sulfur metabolism,signal transduction and cell motility.The metabolic activities of TCS1 might depend on geochemical conditions such as supplies of CO2,hydrogen and sugars,and therefore the TCS1 might be a facultative bacterium in anaerobic saline sediments near cold seeps.

  3. Effect of extract of Floson Chrysanthemi indici on hyperglycemia, hyperipidemia and blood aldose reductase in diabetic KKAy mice%野菊花提取物对KKAy糖尿病小鼠高血糖、高血脂和血醛糖还原酶的影响

    Institute of Scientific and Technical Information of China (English)

    陈雁虹; 张娟; 艾志鹏; 陈婷

    2016-01-01

    OBJECTIVE To investigate the inhibitory effects of extracts of Flos Chrysanthemi indici (FCI) on hyperglycemia,hyperipidemia and aldose reductase (AR) in diabetic KKAy mice. METHODS Twenty-eight male KKAy mice of 8 weeks old were randomly divided into four groups according to the initial fasting glucose:KKAy model group,extract of FCI 20,100 mg · kg-1 groups and glimepiride 0.4 mg · kg-1 group. C57BL/6J mice were taken as the normal control group. These mice were given ig once daily for 7 weeks. The body mass and food intake were recorded weekly. The fasting glucose and OGTT were measured in the last week of the experiment. Mice were sacrificed 1 d after the last admin⁃istration. The indexes of blood biochemistry were determined. Serum insulin,leptin and AR levels were analyzed by immunoassay using ELISA kits. The weights and pathological changes the in the liver,kidney, spleen,including epididymal and perirenal adipose,were measured. The expression of AR mRNA in the kidney was detected by real time-PCR. RESULTS Compared with model control group,the food intake,fasting glucose and AUC in OGTT significantly decreased(P<0.01),the serum glutamic-pyruvic transaminase,glucose,triglycerides,total cholesterol,low-density lipoprotein cholesterol,insulin, leptin and AR levels were markedly reduced(P<0.05),and the organ indexes of the liver and kidney sig⁃nificantly decreased(P<0.05)in extracts of FCI 100 mg · kg-1 group. The histopathological changes in model group included tubular epithelial cell degeneration,hepatic steatosis,islet β-cell degeneration, spleen white pulp atrophy and hypertrophy of the adipocyte. Symptoms listed above were attenuated by extracts of FCI and glimepiride treatment. The inhibitory effects of FCI also inhibited the expression of AR mRNA in the kidney. CONCLUSION The extract of FCI has hypoglycemic and hypolipidemic effect and can inhibit AR in KKAy mice,which may protect the kidney,liver,adipose,pancrea and spleen from the damage of

  4. Effect of Sulfate Reduced Bacterium on Corrosion Behavior of 10CrMoAl Steel

    Institute of Scientific and Technical Information of China (English)

    WANG Hua; LIANG Cheng-hao

    2007-01-01

    The effects of sulfate reduced bacterium (SRB) on the corrosion behavior of 10CrMoAl steel in seawater were studied by chemical immersion, potentiodynamic polarization, electrochemical impedance spectroscopy measurement, and scanning electron microscope techniques. The results show that the content of element sulfur in the corrosion product of 10CrMoAl steel in seawater with SRB is up to 9.23%, which is higher than that of the same in sterile seawater. X-ray diffraction demonstrates that the main corrosion product is FeS. SRB increases the corrosion rate by anodic depolarization of the metabolized sulfide product. SEM observation indicates that the corrosion product is not distributed continuously; in addition, bacilliform sulfate-reduced bacterium accumulates on the local surface of 10CrMoAl steel. Hence, SRB enhances sensitivity to the localized corrosion of 10CrMoAl steel in seawater.

  5. Melanin from the nitrogen-fixing bacterium Azotobacter chroococcum: a spectroscopic characterization.

    Science.gov (United States)

    Banerjee, Aulie; Supakar, Subhrangshu; Banerjee, Raja

    2014-01-01

    Melanins, the ubiquitous hetero-polymer pigments found widely dispersed among various life forms, are usually dark brown/black in colour. Although melanins have variety of biological functions, including protection against ultraviolet radiation of sunlight and are used in medicine, cosmetics, extraction of melanin from the animal and plant kingdoms is not an easy task. Using complementary physicochemical techniques (i.e. MALDI-TOF, FTIR absorption and cross-polarization magic angle spinning solid-state (13)C NMR), we report here the characterization of melanins extracted from the nitrogen-fixing non-virulent bacterium Azotobacter chroococcum, a safe viable source. Moreover, considering dihydroxyindole moiety as the main constituent, an effort is made to propose the putative molecular structure of the melanin hetero-polymer extracted from the bacterium. Characterization of the melanin obtained from Azotobacter chroococcum would provide an inspiration in extending research activities on these hetero-polymers and their use as protective agent against UV radiation.

  6. Economic Game Theory to Model the Attenuation of Virulence of an Obligate Intracellular Bacterium.

    Science.gov (United States)

    Tago, Damian; Meyer, Damien F

    2016-01-01

    Diseases induced by obligate intracellular pathogens have a large burden on global human and animal health. Understanding the factors involved in the virulence and fitness of these pathogens contributes to the development of control strategies against these diseases. Based on biological observations, a theoretical model using game theory is proposed to explain how obligate intracellular bacteria interact with their host. The equilibrium in such a game shows that the virulence and fitness of the bacterium is host-triggered and by changing the host's defense system to which the bacterium is confronted, an evolutionary process leads to an attenuated strain. Although, the attenuation procedure has already been conducted in practice in order to develop an attenuated vaccine (e.g., with Ehrlichia ruminantium), there was a lack of understanding of the theoretical basis behind this process. Our work provides a model to better comprehend the existence of different phenotypes and some underlying evolutionary mechanisms for the virulence of obligate intracellular bacteria.

  7. Isolation and biological characteristics of aerobic marine magnetotactic bacterium YSC-1

    Institute of Scientific and Technical Information of China (English)

    GAO Jun; PAN Hongmiao; YUE Haidong; SONG Tao; ZHAO Yong; CHEN Guanjun; Wu Longfei; XIAO Tian

    2006-01-01

    Magnetotactic bacteria have become a hot spot of research in microbiology attracting intensive interest of researchers in multiple disciplinary fields. However, the studies were limited in few fastidious bacteria. The objective of this study aims at isolating new marine magnetic bacteria and better comprehension of magnetotactic bacteria. In this study, an aerobic magnetotactic bacterium YSC-1 was isolated from sediments in the Yellow Sea Cold Water Mass (YSCWM). In TEM, magnetic cells have one or several circular magnetosomes in dimeter of 100nm, and consist of Fe and Co shown on energy dispersive X-ray spectrum. The biological and physiological characteristics of this bacterium were also described. The colour of YSC-1 colony is white in small rod. The gran stain is negative. Results showed that Strain YSC-1 differs from microaerophile magnetotactic bacteria MS-1 and WD-1 in biology.

  8. The bacterium Xenorhabdus nematophila inhibits phospholipases A2 from insect, prokaryote, and vertebrate sources

    Science.gov (United States)

    Park, Youngjin; Kim, Yonggyun; Stanley, David

    The bacterium, Xenorhabdus nematophila, is a virulent insect pathogen. Part of its pathogenicity is due to impairing cellular immunity by blocking biosynthesis of eicosanoids, the major recognized signal transduction system in insect cellular immunity. X. nematophila inhibits the first step in eicosanoid biosynthesis, phospholipase A2 (PLA2). Here we report that the bacterium inhibits PLA2 from two insect immune tissues, hemocytes and fat body, as well as PLA2s selected to represent a wide range of organisms, including prokaryotes, insects, reptiles, and mammals. Our finding on a bacterial inhibitor of PLA2 activity contributes new insight into the chemical ecology of microbe-host interactions, which usually involve actions rather than inhibitors of PLA2s.

  9. Inflammasomes Coordinate Pyroptosis and Natural Killer Cell Cytotoxicity to Clear Infection by a Ubiquitous Environmental Bacterium.

    Science.gov (United States)

    Maltez, Vivien I; Tubbs, Alan L; Cook, Kevin D; Aachoui, Youssef; Falcone, E Liana; Holland, Steven M; Whitmire, Jason K; Miao, Edward A

    2015-11-17

    Defective neutrophils in patients with chronic granulomatous disease (CGD) cause susceptibility to extracellular and intracellular infections. Microbes must first be ejected from intracellular niches to expose them to neutrophil attack, so we hypothesized that inflammasomes detect certain CGD pathogens upstream of neutrophil killing. Here, we identified one such ubiquitous environmental bacterium, Chromobacterium violaceum, whose extreme virulence was fully counteracted by the NLRC4 inflammasome. Caspase-1 protected via two parallel pathways that eliminated intracellular replication niches. Pyroptosis was the primary bacterial clearance mechanism in the spleen, but both pyroptosis and interleukin-18 (IL-18)-driven natural killer (NK) cell responses were required for liver defense. NK cells cleared hepatocyte replication niches via perforin-dependent cytotoxicity, whereas interferon-γ was not required. These insights suggested a therapeutic approach: exogenous IL-18 restored perforin-dependent cytotoxicity during infection by the inflammasome-evasive bacterium Listeria monocytogenes. Therefore, inflammasomes can trigger complementary programmed cell death mechanisms, directing sterilizing immunity against intracellular bacterial pathogens.

  10. Economic Game Theory to Model the Attenuation of Virulence of an Obligate Intracellular Bacterium

    Science.gov (United States)

    Tago, Damian; Meyer, Damien F.

    2016-01-01

    Diseases induced by obligate intracellular pathogens have a large burden on global human and animal health. Understanding the factors involved in the virulence and fitness of these pathogens contributes to the development of control strategies against these diseases. Based on biological observations, a theoretical model using game theory is proposed to explain how obligate intracellular bacteria interact with their host. The equilibrium in such a game shows that the virulence and fitness of the bacterium is host-triggered and by changing the host's defense system to which the bacterium is confronted, an evolutionary process leads to an attenuated strain. Although, the attenuation procedure has already been conducted in practice in order to develop an attenuated vaccine (e.g., with Ehrlichia ruminantium), there was a lack of understanding of the theoretical basis behind this process. Our work provides a model to better comprehend the existence of different phenotypes and some underlying evolutionary mechanisms for the virulence of obligate intracellular bacteria. PMID:27610355

  11. Regulation of glutamine synthetase activity by adenylylation in the Gram-positive bacterium Streptomyces cattleya.

    Science.gov (United States)

    Streicher, S L; Tyler, B

    1981-01-01

    The enzymatic activity of glutamine synthetase [GS; L-glutamate:ammonia ligase (ADP-forming), EC 6.3.1.2] from the Gram-positive bacterium Streptomyces cattleya is regulated by covalent modification. In whole cells containing high levels of GS the addition of ammonium chloride leads to a rapid decline in GS activity. Crude extracts prepared from such ammonia-shocked cells had very low levels of GS activity as measured by biosynthetic and gamma-glutamyltransferase assays. Incubation of the crude extracts with snake venom phosphodiesterase restored GS activity. In cell extracts, GS was also inactivated by an ATP- and glutamine-dependent reaction. Radioactive labeling studies demonstrated the incorporation of an AmP moiety into GS protein upon modification. Our results suggest a covalent modification of GS in a Gram-positive bacterium. This modification appears to be adenylylation of the GS subunit similar to that found in the Gram-negative bacteria.

  12. A Streamlined Strategy for Biohydrogen Production with Halanaerobium hydrogeniformans, an Alkaliphilic Bacterium.

    Science.gov (United States)

    Begemann, Matthew B; Mormile, Melanie R; Sitton, Oliver C; Wall, Judy D; Elias, Dwayne A

    2012-01-01

    Biofuels are anticipated to enable a shift from fossil fuels for renewable transportation and manufacturing fuels, with biohydrogen considered attractive since it could offer the largest reduction of global carbon budgets. Currently, lignocellulosic biohydrogen production remains inefficient with pretreatments that are heavily fossil fuel-dependent. However, bacteria using alkali-treated biomass could streamline biofuel production while reducing costs and fossil fuel needs. An alkaliphilic bacterium, Halanaerobiumhydrogeniformans, is described that is capable of biohydrogen production at levels rivaling neutrophilic strains, but at pH 11 and hypersaline conditions. H. hydrogeniformans ferments a variety of 5- and 6-carbon sugars derived from hemicellulose and cellulose including cellobiose, and forms the end products hydrogen, acetate, and formate. Further, it can also produce biohydrogen from switchgrass and straw pretreated at temperatures far lower than any previously reported and in solutions compatible with growth. Hence, this bacterium can potentially increase the efficiency and efficacy of biohydrogen production from renewable biomass resources.

  13. A Streamlined Strategy for Biohydrogen Production with Halanaerobium hydrogeniformans, an Alkaliphilic Bacterium

    Directory of Open Access Journals (Sweden)

    Matthew eBegemann

    2012-03-01

    Full Text Available Biofuels are anticipated to enable a shift from fossil fuels for renewable transportation and manufacturing fuels, with biohydrogen considered attractive since it could offer the largest reduction of global carbon budgets. Currently, lignocellulosic biohydrogen production remains inefficient with pretreatments that are heavily fossil fuel-dependent. However, bacteria using alkali-treated biomass could streamline biofuel production while reducing costs and fossil fuel needs. An alkaliphilic bacterium, Halanaerobium hydrogeniformans, is described that is capable of biohydrogen production at levels rivaling neutrophilic strains, but at pH 11 and hypersaline conditions. H. hydrogeniformans ferments a variety of 5- and 6- carbon sugars derived from hemicellulose and cellulose including cellobiose, and forms the end products hydrogen, acetate and formate. Further, it can also produce biohydrogen from switchgrass and straw pretreated at temperatures far lower than any previously reported and in solutions compatible with growth. Hence, this bacterium can potentially increase the efficiency and efficacy of biohydrogen production from renewable biomass resources.

  14. Draft Genome Sequence of Uncultured SAR324 Bacterium lautmerah10, Binned from a Red Sea Metagenome

    KAUST Repository

    Haroon, Mohamed

    2016-02-11

    A draft genome of SAR324 bacterium lautmerah10 was assembled from a metagenome of a surface water sample from the Red Sea, Saudi Arabia. The genome is more complete and has a higher G+C content than that of previously sequenced SAR324 representatives. Its genomic information shows a versatile metabolism that confers an advantage to SAR324, which is reflected in its distribution throughout different depths of the marine water column.

  15. Draft Genome Sequence of the Antitrypanosomally Active Sponge-Associated Bacterium Actinokineospora sp. Strain EG49

    KAUST Repository

    Harjes, Janno

    2014-03-06

    The marine sponge-associated bacterium Actinokineospora sp. strain EG49 produces the antitrypanosomal angucycline-like compound actinosporin A. The draft genome of Actinokineospora sp. EG49 has a size of 7.5 megabases and a GC content of 72.8% and contains 6,629 protein-coding sequences (CDS). antiSMASH predicted 996 genes residing in 36 secondary metabolite gene clusters.

  16. Sexual transmission of a plant pathogenic bacterium, Candidatus Liberibacter asiaticus, between conspecific insect vectors during mating.

    Directory of Open Access Journals (Sweden)

    Rajinder S Mann

    Full Text Available Candidatus Liberibacter asiaticus is a fastidious, phloem-inhabiting, gram-negative bacterium transmitted by Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Psyllidae. The bacterium is the presumed causal agent of huanglongbing (HLB, one of the most destructive and economically important diseases of citrus. We investigated whether Las is transmitted between infected and uninfected D. citri adults during courtship. Our results indicate that Las was sexually transmitted from Las-infected male D. citri to uninfected females at a low rate (<4% during mating. Sexual transmission was not observed following mating of infected females and uninfected males or among adult pairs of the same sex. Las was detected in genitalia of both sexes and also in eggs of infected females. A latent period of 7 days or more was required to detect the bacterium in recipient females. Rod shaped as well as spherical structures resembling Las were observed in ovaries of Las-infected females with transmission electron microscopy, but were absent in ovaries from uninfected D. citri females. The size of the rod shaped structures varied from 0.39 to 0.67 µm in length and 0.19 to 0.39 µm in width. The spherical structures measured from 0.61 to 0.80 µm in diameter. This investigation provides convincing evidence that a plant pathogenic bacterium is sexually transmitted from male to female insects during courtship and established evidence that bacteria persist in reproductive organs. Moreover, these findings provide an alternative sexually horizontal mechanism for the spread of Las within populations of D. citri, even in the absence of infected host trees.

  17. Five new amicoumacins isolated from a marine-derived Bacterium bacillus subtilis

    KAUST Repository

    Li, Yongxin

    2012-02-03

    Four novel amicoumacins, namely lipoamicoumacins A-D (1-4), and one new bacilosarcin analog (5) were isolated from culture broth of a marine-derived bacterium Bacillus subtilis, together with six known amicoumacins. Their structures were elucidated on the basis of extensive spectroscopic (2D NNR, IR, CD and MS) analysis and in comparison with data in literature. 2012 by the authors; licensee MDPI.

  18. Insights in Nanoparticle-Bacterium Interactions: New Frontiers to Bypass Bacterial Resistance to Antibiotics.

    Science.gov (United States)

    Diab, Roudayna; Khameneh, Bahman; Joubert, Olivier; Duval, Raphael

    2015-01-01

    Nanotechnology has been revealed as a fundamental approach for antibiotics delivery. In this paper, recent findings demonstrating the superiority of nanocarried-antibiotics over "naked" ones and the ways by which nanoparticles can help to overwhelm bacterial drug resistance are reviewed. The second part of this paper sheds light on nanoparticle-bacterium interaction patterns. Finally, key factors affecting the effectiveness of nanoparticles interactions with bacteria are discussed.

  19. Two New Cholic Acid Derivatives from the Marine Ascidian-Associated Bacterium Hasllibacter halocynthiae

    Directory of Open Access Journals (Sweden)

    Sung Hun Kim

    2012-10-01

    Full Text Available The investigation of secondary metabolites in liquid cultures of a recently discovered marine bacterium, Hasllibacter halocynthiae strain KME 002T, led to the isolation of two new cholic acid derivatives. The structures of these compounds were determined to be 3,3,12-trihydroxy-7-ketocholanic acid (1 and 3,3,12-trihydroxy-7-deoxycholanic acid (2 through HRFABMS and NMR data analyses.

  20. Degradation of p-nitrophenol by the phototrophic bacterium Rhodobacter capsulatus.

    Science.gov (United States)

    Roldán, M D; Blasco, R; Caballero, F J; Castillo, F

    1998-01-01

    The phototrophic bacterium Rhodobacter capsulatus detoxified p-nitrophenol and 4-nitrocatechol. The bacterium tolerated moderate concentrations of p-nitrophenol (up to 0.5 mM) and degraded it under light at an optimal O2 pressure of 20 kPa. The bacterium did not metabolize the xenobiotic in the dark or under strictly anoxic conditions or high O2 pressure. Bacterial growth with acetate in the presence of p-nitrophenol took place with the simultaneous release of nonstoichiometric amounts of 4-nitrocatechol, which can also be degraded by the bacterium. Crude extracts from R. capsulatus produced 4-nitrocatechol from p-nitrophenol upon the addition of NAD(P)H, although at a very low rate. A constitutive catechol 1, 2-dioxygenase activity yielding cis,cis-muconate was also detected in crude extracts of R. capsulatus. Further degradation of 4-nitrocatechol included both nitrite- and CO2-releasing steps since: (1) a strain of R. capsulatus (B10) unable to assimilate nitrate and nitrite released nitrite into the medium when grown with p-nitrophenol or 4-nitrocatechol, and the nitrite concentration was stoichiometric with the 4-nitrocatechol degraded, and (2) cultures of R. capsulatus growing microaerobically produced low amounts of 14CO2 from radiolabeled p-nitrophenol. The radioactivity was also incorporated into cellular compounds from cells grown with uniformly labeled 14C-p-nitrophenol. From these results we concluded that the xenobiotic is used as a carbon source by R. capsulatus, but that only the strain able to assimilate nitrite (E1F1) can use p-nitrophenol as a nitrogen source.

  1. Identifying the assembly pathway of cyanophage inside the marine bacterium using electron cryo-tomography

    Directory of Open Access Journals (Sweden)

    Wei Dai

    2014-01-01

    Full Text Available Advances in electron cryo-tomography open up a new avenue to visualize the 3-D internal structure of a single bacterium before and after its infection by bacteriophages in its native environment, without using chemical fixatives, fluorescent dyes or negative stains. Such direct observation reveals the presence of assembly intermediates of the bacteriophage and thus allows us to map out the maturation pathway of the bacteriophage inside its host.

  2. Identifying the assembly pathway of cyanophage inside the marine bacterium using electron cryo-tomography.

    Science.gov (United States)

    Dai, Wei; Schmid, Michael F; King, Jonathan A; Chiu, Wah

    2014-06-01

    Advances in electron cryo-tomography open up a new avenue to visualize the 3-D internal structure of a single bacterium before and after its infection by bacteriophages in its native environment, without using chemical fixatives, fluorescent dyes or negative stains. Such direct observation reveals the presence of assembly intermediates of the bacteriophage and thus allows us to map out the maturation pathway of the bacteriophage inside its host.

  3. Identifying the assembly pathway of cyanophage inside the marine bacterium using electron cryo-tomography

    OpenAIRE

    Wei Dai; Schmid, Michael F.; King, Jonathan A.; Wah Chiu

    2014-01-01

    Advances in electron cryo-tomography open up a new avenue to visualize the 3-D internal structure of a single bacterium before and after its infection by bacteriophages in its native environment, without using chemical fixatives, fluorescent dyes or negative stains. Such direct observation reveals the presence of assembly intermediates of the bacteriophage and thus allows us to map out the maturation pathway of the bacteriophage inside its host.

  4. Draft Genome Sequence of Agarivorans albus Strain MKT 106T, an Agarolytic Marine Bacterium.

    Science.gov (United States)

    Yasuike, Motoshige; Nakamura, Yoji; Kai, Wataru; Fujiwara, Atushi; Fukui, Youhei; Satomi, Masataka; Sano, Motohiko

    2013-07-18

    Agarivorans albus is a Gram-negative, strictly aerobic, and agar-hydrolyzing marine bacterium. We present the draft genome sequence of the A. albus strain MKT 106(T), which is composed of 67 contigs (>500 bp) totaling 4,734,285 bp and containing 4,397 coding DNA sequences (CDSs), four rRNAs, and 64 tRNA sequences.

  5. Genome Sequence of Marine Bacterium Idiomarina sp. Strain 28-8, Isolated from Korean Ark Shells.

    Science.gov (United States)

    Kim, Woo-Jin; Kim, Young-Ok; Kim, Dong-Gyun; Nam, Bo-Hye; Kong, Hee Jeong; Jung, Hyungtaek; Lee, Sang-Jun; Kim, Dong-Wook; Kim, Dae-Soo; Chae, Sung-Hwa

    2013-10-03

    Idiomarina sp. strain 28-8 is an aerobic, Gram-negative, flagellar bacterium isolated from the bodies of ark shells (Scapharca broughtonii) collected from underwater sediments in Gangjin Bay, South Korea. Here, we present the draft genome sequence of Idiomarina sp. 28-8 (2,971,606 bp, with a G+C content of 46.9%), containing 2,795 putative coding sequences.

  6. Cadmium resistance and uptake by bacterium, Salmonella enterica 43C, isolated from industrial effluent.

    Science.gov (United States)

    Khan, Zaman; Rehman, Abdul; Hussain, Syed Z; Nisar, Muhammad A; Zulfiqar, Soumble; Shakoori, Abdul R

    2016-12-01

    Cadmium resistant bacterium, isolated from industrial wastewater, was characterized as Salmonella enterica 43C on the basis of biochemical and 16S rRNA ribotyping. It is first ever reported S. enterica 43C bared extreme resistance against heavy metal consortia in order of Pb(2+)>Cd(2+)>As(3+)>Zn(2+)>Cr(6+)>Cu(2+)>Hg(2+). Cd(2+) stress altered growth pattern of the bacterium in time dependent manner. It could remove nearly 57 % Cd(2+) from the medium over a period of 8 days. Kinetic and thermodynamic studies based on various adsorption isotherm models (Langmuir and Freundlich) depicted the Cd(2+) biosorption as spontaneous, feasible and endothermic in nature. Interestingly, the bacterium followed pseudo first order kinetics, making it a good biosorbent for heavy metal ions. The S. enterica 43C Cd(2+) processivity was significantly influenced by temperature, pH, initial Cd(2+) concentration, biomass dosage and co-metal ions. FTIR analysis of the bacterium revealed the active participation of amide and carbonyl moieties in Cd(2+) adsorption confirmed by EDX analysis. Electron micrographs beckoned further surface adsorption and increased bacterial size due to intracellular Cd(2+) accumulation. An overwhelming increase in glutathione and other non-protein thiols levels played a significant role in thriving oxidative stress generated by metal cations. Presence of metallothionein clearly depicted the role of such proteins in bacterial metal resistance mechanism. The present study results clearly declare S. enterica 43C a suitable candidate for green chemistry to bioremediate environmental Cd(2+).

  7. Biological control and endophytism of the olive root bacterium Pseudomonas fluorescens PICF7

    OpenAIRE

    Maldonado González, Mercedes

    2015-01-01

    Olive (Olea europaea L.) has always been a fundamental crop in the Mediterranean Basin. Driven by the fact, among others, that an increasing number of scientific reports highlight the benefits that olive oil consumption has for human health, olive tree cultivation has spread worldwide to other regions with Mediterranean-type climate. Two relevant pathogens affecting olive trees are the hemibiotrophic soil-borne fungus Verticillium dahliae and the bacterium Pseudomonas savastano...

  8. Draft Genome Sequence of the Cyanide-Utilizing Bacterium Pseudomonas fluorescens Strain NCIMB 11764

    OpenAIRE

    2012-01-01

    We report here the 6.97-Mb draft genome sequence of Pseudomonas fluorescens strain NCIMB 11764, which is capable of growth on cyanide as the sole nitrogen source. The draft genome sequence allowed the discovery of several genes implicated in enzymatic cyanide turnover and provided additional information contributing to a better understanding of this organism's unique cyanotrophic ability. This is the first sequenced genome of a cyanide-assimilating bacterium.

  9. Draft genome sequence of the cyanide-utilizing bacterium Pseudomonas fluorescens strain NCIMB 11764.

    Science.gov (United States)

    Vilo, Claudia A; Benedik, Michael J; Kunz, Daniel A; Dong, Qunfeng

    2012-12-01

    We report here the 6.97-Mb draft genome sequence of Pseudomonas fluorescens strain NCIMB 11764, which is capable of growth on cyanide as the sole nitrogen source. The draft genome sequence allowed the discovery of several genes implicated in enzymatic cyanide turnover and provided additional information contributing to a better understanding of this organism's unique cyanotrophic ability. This is the first sequenced genome of a cyanide-assimilating bacterium.

  10. Permanent draft genome of the malachite-green-tolerant bacterium Rhizobium sp. MGL06.

    Science.gov (United States)

    Liu, Yang; Wang, Runping; Zeng, Runying

    2014-12-01

    Rhizobium sp. MGL06, the first Rhizobium isolate from a marine environment, is a malachite-green-tolerant bacterium with a broader salinity tolerance (range: 0.5% to 9%) than other rhizobia. This study sequences and annotates the draft genome sequence of this strain. Genome sequence information provides a basis for analyzing the malachite green tolerance, broad salinity adaptation, nitrogen fixation properties, and taxonomic classification of the isolate.

  11. Complete genome sequence of Rufibacter tibetensis strain 1351, a radiation-resistant bacterium from Tibet plateau.

    Science.gov (United States)

    Zhang, Yi; Yu, Can; Zhou, Mengzhou; Tang, Jingfeng; Li, Xin; Wang, Zhi; Li, Zhijun; Yao, Juan; Li, Pei; Zheng, Guobin; Chen, Xiong; Dai, Jun

    2015-12-20

    Rufibacter tibetensis strain 1351, isolated from the soil of the Tibet plateau of China, belongs to the family of Cytophagaceae. It is a red-pigmented, gram-negative, strictly aerobic and rod-shaped bacterium and shows resistance to UV radiation. Here, we report its complete genome sequence, which can help us find the key genes of the carotenoid biosynthesis and resistance to UV radiation.

  12. Fourier transform infrared spectroscopic study of intact cells of the nitrogen-fixing bacterium Azospirillum brasilense

    Science.gov (United States)

    Kamnev, A. A.; Ristić, M.; Antonyuk, L. P.; Chernyshev, A. V.; Ignatov, V. V.

    1997-06-01

    The data of Fourier transform infrared (FTIR) spectroscopic measurements performed on intact cells of the soil nitrogen-fixing bacterium Azospirillum brasilense grown in a standard medium and under the conditions of an increased metal uptake are compared and discussed. The structural FTIR information obtained is considered together with atomic absorption spectrometry (AAS) data on the content of metal cations in the bacterial cells. Some methodological aspects concerning preparation of bacterial cell samples for FTIR measurements are also discussed.

  13. Draft Genome Sequence of Sphingobium ummariense Strain RL-3, a Hexachlorocyclohexane-Degrading Bacterium.

    Science.gov (United States)

    Kohli, Puneet; Dua, Ankita; Sangwan, Naseer; Oldach, Phoebe; Khurana, J P; Lal, Rup

    2013-11-14

    Here, we report the draft genome sequence of the hexachlorocyclohexane (HCH)-degrading bacterium Sphingobium ummariense strain RL-3, which was isolated from the HCH dumpsite located in Lucknow, India (27°00'N and 81°09'E). The annotated draft genome sequence (4.75 Mb) of strain RL-3 consisted of 139 contigs, 4,645 coding sequences, and 65% G+C content.

  14. Bacillus marcorestinctum sp. nov., a Novel Soil Acylhomoserine Lactone Quorum-Sensing Signal Quenching Bacterium

    OpenAIRE

    Xianzhen Li; Bo Zhu; Nuo Li; Fang Chen; Yan Han

    2010-01-01

    A Gram-positive, facultatively anaerobic, endospore-forming and rod-shaped bacterium was isolated from soil samples and designated strain LQQ. This organism strongly quenches the acylhomoserine lactone quorum-sensing signal. The LQQ strain exhibits phenotypic characteristics consistent with its classification in the genus Bacillus. It is positive in catalase and no special growth factor is needed. It uses glucose as sole carbon source. The DNA G + C content is 39.8 mol %. The closest relative...

  15. Genetic Engineering of a Radiation-Resistant Bacterium for Biodegradation of Mixed Wastes--Final Report

    Energy Technology Data Exchange (ETDEWEB)

    Mary E. Lidstrom

    2003-12-26

    Aqueous mixed low level wastes (MLLW) containing radionuclides, solvents, and/or heavy metals represent a serious current and future problem for DOE environmental management and cleanup. In order to provide low-cost treatment alternatives under mild conditions for such contained wastes, we have proposed to use the radiation-resistant bacterium, Deinococcus radiodurans. This project has focused on developing D. radiodurans strains for dual purpose processes: cometabolic treatment of haloorganics and other solvents and removal of heavy metals from waste streams in an above-ground reactor system. The characteristics of effective treatment strains that must be attained are: (a) high biodegradative and metal binding activity; (b) stable treatment characteristics in the absence of selection and in the presence of physiological stress; (c) survival and activity under harsh chemical conditions, including radiation. The result of this project has been a suite of strains with high biodegradative capabilities that are candidates for pilot stage treatment systems. In addition, we have determined how to create conditions to precipitate heavy metals on the surface of the bacterium, as the first step towards creating dual-use treatment strains for contained mixed wastes of importance to the DOE. Finally, we have analyzed stress response in this bacterium, to create the foundation for developing treatment processes that maximize degradation while optimizing survival under high stress conditions.

  16. Antimicrobial activity and biosynthesis of nanoparticles by endophytic bacterium inhabiting Coffee arabica L.

    Directory of Open Access Journals (Sweden)

    Syed Baker

    2012-12-01

    Full Text Available The interface between endophytes and nanomaterials is a relatively new and unexplored area the present study evaluates screening of bacterial endophytes from surfaced sterilized leaf and stem segments of agro economical plant Coffee arabica L. towards synthesis of silver nanoparticles and antimicrobial metabolites. Among thirty two endophytes isolated nine isolates exhibited antimicrobial activity among which one bacterium was capable of extracellular synthesis of silver nanoparticles upon evaluation of supernatant with 1 mM of silver nitrate, biosynthesis of silver nanoparticles were assessed by UV-Visible Spectroscopy and the bacterium was capable of secreting antimicrobial secondary metabolites upon crude ethyl acetate extract evaluated for antimicrobial activity against panel of both gram positive and gram negative as well as phytopathogenic fungi. Partial characterization was carried out via bioautographic technique with Rf value 0.3 and 0.6 exhibiting antimicrobial activity against MRSA strain. Further studies in this area will be promising enough for molecular characterization of endophytic bacterium and chemical profiling of antimicrobial metabolites at the same time physiochemical characterization of nanoparticles will be valuable to reveal the size and shape. 

  17. Antagonism and Molecular Identification of an Antibiotic Bacterium BS04 Against Phytopathogenic Fungi and Bacteria

    Institute of Scientific and Technical Information of China (English)

    Xie Jing(谢晶); Ge Shaorong; Tao Yong; Gao Ping; Liu Kun; Liu Shigui

    2004-01-01

    Through a modified agar well diffusion assay, antagonism of bacterium BS04 is tested. The data show that BS04 has antibiotic activity against phytopathogenic fungi and bacteria, including Phoma wasabiae Yokogi, Cochlibolus Heterostrophu, Exserohilum Turcicum, Curuvularia Lunata (Walk) Boed, Thantephorus cucumris, Fusarium graminearum, Xanthomonas axonopodis pv. Citri (Hasse) Dye and Xanthomonas zingiberi (Uyeda) Savulescu. The products of bacterium BS04 can endure the treatment of a wide range of pH, and maintain the antibiotic activity after treatment of 100℃ for 30 min. The result suggests that bacterium BS04 has the potential as a promising biocontrol agent. In order to determine the taxonomic placement, the molecular identification of BS04 is performed. The comparative analysis of 16s rDNA sequences indicates that the 16s rDNA sequence of BS04 is highly homologous with sequences of typical Paenibacillus bacteria from the RPD library (from 92% to 99%). And the constructed phylogenetic tree by using maximum-likelihood method with Bootstrap Trial 1000 proves that BS04 is subjected to Paenibacillus polymyxa.

  18. Widespread association of a Rickettsiales-like bacterium with reef-building corals.

    Science.gov (United States)

    Casas, Veronica; Kline, David I; Wegley, Linda; Yu, Yanan; Breitbart, Mya; Rohwer, Forest

    2004-11-01

    White band disease type I (WBD I) has been a major cause of the dramatic decline of Acroporid coral populations throughout the Caribbean during the last two decades, yet the aetiological agent of this disease is unknown. In this study, the bacterial communities associated with both healthy and diseased Acropora species were compared by 16S rDNA analyses. The bacterial communities of both healthy and diseased Acropora spp. were dominated by a single ribotype with 90% identity to a bacterium in the order Rickettsiales. Screening by nested PCR specific to the coral-associated Rickettsiales 1 (CAR1) bacterium showed that this microbe was widespread in both healthy and diseased A. cervicornis and A. palmata corals from 'healthy' (i.e. low WBD I incidence) and 'stressed' reefs (i.e. high WBD I incidence). These results indicate that there were no dramatic changes in the composition of the microbial community associated with WBD I. CAR1 was also associated with non-Acroporid corals of the Caribbean, as well as with two Acroporid corals native to the Pacific. CAR1 was not present in the water column. This bacterium was also absent from preserved Caribbean Acroporid samples collected between 1937 and 1980 before the outbreak of WBD I. These results suggest CAR1 is a relatively new bacterial associate of Acroporids and that a non-bacterial pathogen might be the cause of WBD I.

  19. Rhodococcus sp. Q5, a novel agarolytic bacterium isolated from printing and dyeing wastewater.

    Science.gov (United States)

    Feng, Zehua; Peng, Lin; Chen, Mei; Li, Mengying

    2012-09-01

    An agar-degrading bacterium, Rhodococcus sp. Q5, was isolated from printing and dyeing wastewater using a mineral salts agar plate containing agar as the sole carbon source. The bacterium grew from pH 4.0 to 9.0, from 15 to 35°C, and in NaCl concentrations of 0-5 %; optimal values were pH 6.0, 30°C, and 1 % NaCl. Maximal agarase production was observed at pH 6.0 and 30°C. The bacterium did not require NaCl for growth or agarase production. The agarase secreted by Q5 was inducible by agar and was repressed by all simple sugars tested except lactose. Strain Q5 could hydrolyze starch but not cellulose or carboxymethyl cellulose. Agarase activity could also be detected in the medium when lactose or starch was the sole source of carbon and energy. Strain Q5 could grow in nitrogen-free mineral media; an organic nitrogen source was more effective than inorganic carbon sources for growth and agarase production. Addition of more organic nitrogen (peptone) to the medium corresponded with reduced agarase activity.

  20. Enhancement of survival and electricity production in an engineered bacterium by light-driven proton pumping.

    Science.gov (United States)

    Johnson, Ethan T; Baron, Daniel B; Naranjo, Belén; Bond, Daniel R; Schmidt-Dannert, Claudia; Gralnick, Jeffrey A

    2010-07-01

    Microorganisms can use complex photosystems or light-dependent proton pumps to generate membrane potential and/or reduce electron carriers to support growth. The discovery that proteorhodopsin is a light-dependent proton pump that can be expressed readily in recombinant bacteria enables development of new strategies to probe microbial physiology and to engineer microbes with new light-driven properties. Here, we describe functional expression of proteorhodopsin and light-induced changes in membrane potential in the bacterium Shewanella oneidensis strain MR-1. We report that there were significant increases in electrical current generation during illumination of electrochemical chambers containing S. oneidensis expressing proteorhodopsin. We present evidence that an engineered strain is able to consume lactate at an increased rate when it is illuminated, which is consistent with the hypothesis that proteorhodopsin activity enhances lactate uptake by increasing the proton motive force. Our results demonstrate that there is coupling of a light-driven process to electricity generation in a nonphotosynthetic engineered bacterium. Expression of proteorhodopsin also preserved the viability of the bacterium under nutrient-limited conditions, providing evidence that fulfillment of basic energy needs of organisms may explain the widespread distribution of proteorhodopsin in marine environments.

  1. Epidemiological analysis of acute diarrhea in children and inspection of pathogenic bacterium, viruses and other microorganisms

    Institute of Scientific and Technical Information of China (English)

    Li Hu; Yan Wang

    2016-01-01

    Objective:To investigate of epidemiological analysis of acute diarrhea in children, and to discuss the inspection of pathogenic bacterium, viruses and other microorganisms, in order to provide theoretical basis for the prevention and treatment of the disease.Methods: Five hundred and sixty-two cases of children with acute diarrhea treated in our center were selected as the research subjects, whose epidemiological data were analyzed. The fecal samples were collected for bacterial culture and identification, and the distribution characteristics of pathogenic bacteria were collected, then their relative characteristics were analyzed.Results:Children with acute diarrhea were more common in men aged 1-2 years old,and the incidence of time was more concentrated in June-August. There were four hundred and eighty-nine strains in the five hundred and sixty-two cases of children, among which the rate of viruses was the most, and the human rotavirus accounted for 30.67%, and the Shigella bacterium accounted for 20.65% in the total microorganisms, which was the highest detection rate of pathogenic bacterium. Rotavirus infection occured mainly in Winter, but the bacterial and goblet viral diarrhea was prevalent in summer.Conclusions:Children with acute diarrhea were more common in men aged 1-2 years old , and the rate of viruses in the detection of microorganisms is the highest, so targeted treatment should be taken according to the type of infection.

  2. Development of a markerless deletion system for the fish-pathogenic bacterium Flavobacterium psychrophilum.

    Science.gov (United States)

    Gómez, Esther; Álvarez, Beatriz; Duchaud, Eric; Guijarro, José A

    2015-01-01

    Flavobacterium psychrophilum is a Gram-negative fish pathogen that causes important economic losses in aquaculture worldwide. Although the genome of this bacterium has been determined, the function and relative importance of genes in relation to virulence remain to be established. To investigate their respective contribution to the bacterial pathogenesis, effective tools for gene inactivation are required. In the present study, a markerless gene deletion system has been successfully developed for the first time in this bacterium. Using this method, the F. psychrophilum fcpB gene, encoding a predicted cysteine protease homologous to Streptococcus pyogenes streptopain, was deleted. The developed system involved the construction of a conjugative plasmid that harbors the flanking sequences of the fcpB gene and an I-SceI meganuclease restriction site. Once this plasmid was integrated in the genome by homologous recombination, the merodiploid was resolved by the introduction of a plasmid expressing I-SceI under the control of the fpp2 F. psychrophilum inducible promoter. The resulting deleted fcpB mutant presented a decrease in extracellular proteolytic activity compared to the parental strain. However, there were not significant differences between their LD50 in an intramuscularly challenged rainbow trout infection model. The mutagenesis approach developed in this work represents an improvement over the gene inactivation tools existing hitherto for this "fastidious" bacterium. Unlike transposon mutagenesis and gene disruption, gene markerless deletion has less potential for polar effects and allows the mutation of virtually any non-essential gene or gene clusters.

  3. Development of a markerless deletion system for the fish-pathogenic bacterium Flavobacterium psychrophilum.

    Directory of Open Access Journals (Sweden)

    Esther Gómez

    Full Text Available Flavobacterium psychrophilum is a Gram-negative fish pathogen that causes important economic losses in aquaculture worldwide. Although the genome of this bacterium has been determined, the function and relative importance of genes in relation to virulence remain to be established. To investigate their respective contribution to the bacterial pathogenesis, effective tools for gene inactivation are required. In the present study, a markerless gene deletion system has been successfully developed for the first time in this bacterium. Using this method, the F. psychrophilum fcpB gene, encoding a predicted cysteine protease homologous to Streptococcus pyogenes streptopain, was deleted. The developed system involved the construction of a conjugative plasmid that harbors the flanking sequences of the fcpB gene and an I-SceI meganuclease restriction site. Once this plasmid was integrated in the genome by homologous recombination, the merodiploid was resolved by the introduction of a plasmid expressing I-SceI under the control of the fpp2 F. psychrophilum inducible promoter. The resulting deleted fcpB mutant presented a decrease in extracellular proteolytic activity compared to the parental strain. However, there were not significant differences between their LD50 in an intramuscularly challenged rainbow trout infection model. The mutagenesis approach developed in this work represents an improvement over the gene inactivation tools existing hitherto for this "fastidious" bacterium. Unlike transposon mutagenesis and gene disruption, gene markerless deletion has less potential for polar effects and allows the mutation of virtually any non-essential gene or gene clusters.

  4. Programmed cell death in Laminaria japonica (Phaeophyta) tissues infected with alginic acid decomposing bacterium

    Institute of Scientific and Technical Information of China (English)

    WANG Gaoge; LIN Wei; ZHANG Lijing; YAN Xiaojun; DUAN Delin

    2004-01-01

    TdT-mediated dUTP-biotin nick end labeling (TUNEL) is a sensitive and valid method for detecting DNA cleavage in programmed cell death (PCD). Using this method, DNA cleavage was observed in Laminaria japonica sporophytic tissues, which were infected with alginic acid decomposing bacterium. It was found that DNA cleavage occurred 5 min after the infection, the fragments with 3′-OH groups of cleaved nuclear DNA increased with time of infection and spread from the infection site. Although no typical DNA ladder (200 bp/180 bp) was detected by routine agarose gel electrophoresis, the cleavage of nuclear DNA fragments of 97~48.5 kb could be detected by pulsed field gel electrophoresis (PFGE). By using CaspGLOWTM fluorescein active caspase-3 staining method, caspase-3 activity has been detected in response to the infection of alginic acid decomposing bacterium. Our results are similar to the observations in hypersensitive response (HR) of higher plant, suggesting that the rapid cell death of L. Japonica infected by alginic acid decomposing bacterium might be involved in PCD, and indicating that the occurrence of PCD is an active defense process against the pathogen's infection.

  5. Phosphate enhances levan production in the endophytic bacterium Gluconacetobacter diazotrophicus Pal5

    Science.gov (United States)

    Idogawa, Nao; Amamoto, Ryuta; Murata, Kousaku; Kawai, Shigeyuki

    2014-01-01

    Gluconacetobacter diazotrophicus is a gram-negative and endophytic nitrogen-fixing bacterium that has several beneficial effects in host plants; thus, utilization of this bacterium as a biofertilizer in agriculture may be possible. G. diazotrophicus synthesizes levan, a D-fructofuranosyl polymer with β-(2→6) linkages, as an exopolysaccharide and the synthesized levan improves the stress tolerance of the bacterium. In this study, we found that phosphate enhances levan production by G. diazotrophicus Pal5, a wild type strain that showed a stronger mucous phenotype on solid medium containing 28 mM phosphate than on solid medium containing 7 mM phosphate. A G. diazotrophicus Pal5 levansucrase disruptant showed only a weak mucous phenotype regardless of the phosphate concentration, indicating that the mucous phenotype observed on 28 mM phosphate medium was caused by levan. To our knowledge, this is the first report of the effect of a high concentration of phosphate on exopolysaccharide production. PMID:24717418

  6. Anomalous magnetic orientations of magnetosome chains in a magnetotactic bacterium: Magnetovibrio blakemorei strain MV-1.

    Directory of Open Access Journals (Sweden)

    Samanbir S Kalirai

    Full Text Available There is a good deal of published evidence that indicates that all magnetosomes within a single cell of a magnetotactic bacterium are magnetically oriented in the same direction so that they form a single magnetic dipole believed to assist navigation of the cell to optimal environments for their growth and survival. Some cells of the cultured magnetotactic bacterium Magnetovibrio blakemorei strain MV-1 are known to have relatively wide gaps between groups of magnetosomes that do not seem to interfere with the larger, overall linear arrangement of the magnetosomes along the long axis of the cell. We determined the magnetic orientation of the magnetosomes in individual cells of this bacterium using Fe 2p X-ray magnetic circular dichroism (XMCD spectra measured with scanning transmission X-ray microscopy (STXM. We observed a significant number of cases in which there are sub-chains in a single cell, with spatial gaps between them, in which one or more sub-chains are magnetically polarized opposite to other sub-chains in the same cell. These occur with an estimated frequency of 4.0±0.2%, based on a sample size of 150 cells. We propose possible explanations for these anomalous cases which shed insight into the mechanisms of chain formation and magnetic alignment.

  7. An oleaginous bacterium that intrinsically accumulates long-chain free Fatty acids in its cytoplasm.

    Science.gov (United States)

    Katayama, Taiki; Kanno, Manabu; Morita, Naoki; Hori, Tomoyuki; Narihiro, Takashi; Mitani, Yasuo; Kamagata, Yoichi

    2014-02-01

    Medium- and long-chain fatty acids are present in organisms in esterified forms that serve as cell membrane constituents and storage compounds. A large number of organisms are known to accumulate lipophilic materials as a source of energy and carbon. We found a bacterium, designated GK12, that intrinsically accumulates free fatty acids (FFAs) as intracellular droplets without exhibiting cytotoxicity. GK12 is an obligatory anaerobic, mesophilic lactic acid bacterium that was isolated from a methanogenic reactor. Phylogenetic analysis based on 16S rRNA gene sequences showed that GK12 is affiliated with the family Erysipelotrichaceae in the phylum Firmicutes but is distantly related to type species in this family (less than 92% similarity in 16S rRNA gene sequence). Saturated fatty acids with carbon chain lengths of 14, 16, 18, and 20 were produced from glucose under stress conditions, including higher-than-optimum temperatures and the presence of organic solvents that affect cell membrane integrity. FFAs were produced at levels corresponding to up to 25% (wt/wt) of the dry cell mass. Our data suggest that FFA accumulation is a result of an imbalance between excess membrane fatty acid biosynthesis due to homeoviscous adaptation and limited β-oxidation activity due to anaerobic growth involving lactic acid fermentation. FFA droplets were not further utilized as an energy and carbon source, even under conditions of starvation. A naturally occurring bacterium that accumulates significant amounts of long-chain FFAs with noncytotoxicity would provide useful strategies for microbial biodiesel production.

  8. Detection of Erwinia amylovora in imported apple fruit with nested PCR%进境苹果果实中梨火疫病菌的套式PCR检测

    Institute of Scientific and Technical Information of China (English)

    贾平乔; 周国梁; 吴杏霞; 张卫东; 杨晓君; 徐殿胜; 易建平

    2009-01-01

    针对进境商用苹果果实携带梨火疫病菌Erwinia amylovora数量有限的特点,选取源于病菌pEA29质粒的2对引物P29A/P29B和PEANT1/PEANT2配对组合成套式PCR,其检测灵敏度可达0.15 Pg菌体DNA,检测灵敏度高于EPPO推荐的单管套式PCR方法和常规PCR方法.分别利用这3种PCR检测方法对美国、新西兰、日本和智利等国进境的166批苹果样品进行检测,3种检测方法的样品阳性率分别为53.6%、38.0%和8.4%,试验结果表明此套式PCR检测方法可用于进境商用苹果的梨火疫病菌快速检测.进境样品的检测结果证实了进境商用苹果果实中存在梨火疫病菌的可能性.%The specific primers P29A/P29B and PEANT1/PEANT2 derived from plasmid pEA29 of Erwinia amylovora were selected to combine nested PCR method for the detection of trace E. amylovora in imported apple fruits with a limit as low as 0. 15 pg of total DNA. From 166 samples of imported apple fruits from USA, Japan, New Zealand and Chile, this detection method achieved better results than nested PCR in a sin-gle closed tube that EPPO recommended and standard PCR with 53.6% , 38.0% and 8. 4% of positive sam-ples, respectively. This nested PCR method was proposed to be used for the detection of E. amylovora in im-ported commercial apple fruits. The results of detection verified the possibility of imported commercial apple fruits harbouring E. amylovora in the calyxes and fruit stalks.

  9. The field trials of neomycin against Erwinia carotovora subsp, carotovora%新霉素对大白菜软腐病的田间药效试验研究

    Institute of Scientific and Technical Information of China (English)

    王东磊; 达娃; 崔文华; 陈龙; 侯太平

    2011-01-01

    The stability of neomycin was tested by physical and chemical factors to study the antibacterial activity. Field trials of neomycin against Erwinia carotovora subsp. Carotovora was conducted to study neomycin control effect. Physical and chemical factors test showed that neomycin is very stable, it could resist strong acid, alkali, high temperature and UV light. Besides neomycin has many advantages in producing, storing, applying. The results of field trial show that neomycin has good control efficiency a-gainst Erwinia carotovora subsp. Carotovorat. The control effects of neomycin with 200 mg/L,100 mg/ L、50 mg/L were 84.1%,72. 83% and 63. 2% respectively. The control effect of streptomycin with 200 mg/L was 60. 24%. The test results show that neomycin has a promising prospect in pesticide industry. All these advantages make it a very good candidate for pestcide.%本研究通过各种理化因子对新霉素活性的影响考察新霉素的稳定性,进一步通过田间小区试验方法考察新霉素对大白菜软腐病的田间实际防治效果.理化因子试验结果表明,新霉素具有良好的稳定性,能够耐酸碱,耐高温、耐紫外线的直射,具有生产、储藏和应用上的优势.田间小区试验结果表明,新霉素200 mg/L、100 mg/L、50 mg/L的防治效果均优于常用药剂72%农用硫酸链霉素.新霉素200 mg/L、100 mg/L、50 mg/L的防治效果为84.1%、72.83%、63.2%.72%农用硫酸链霉素50mg/L的防治效果为60.24%.两组试验显示了新霉素具有良好的应用前景,为把新霉素开发为一种新型的抗生素类农药奠定了基础.

  10. Comparative proteomics and activity of a green sulfur bacterium across the water column of Lake Cadagno, Switzerland

    DEFF Research Database (Denmark)

    Habicht, Kirsten Silvia; Miller, Mette; Cox, Raymond Pickett

    2011-01-01

    Primary production in the meromictic Lake Cadagno, Switzerland, is dominated by anoxygenic photosynthesis. The green sulfur bacterium Chlorobium clathratiforme is the dominant phototrophic organism in the lake, comprising more than half of the bacterial population, and its biomass increases 3...

  11. Genome Sequence of the Endophytic Bacterium Bacillus thuringiensis Strain KB1, a Potential Biocontrol Agent against Phytopathogens

    OpenAIRE

    Jeong, Haeyoung; Jo, Sung Hee; Hong, Chi Eun; Park, Jeong Mee

    2016-01-01

    Bacillus thuringiensis is the most widely known microbial pesticide used in agricultural applications. Herein, we report a draft genome sequence of the endophytic bacterium Bacillus thuringiensis strain KB1, which exhibits antagonism against phytopathogens.

  12. Genome Sequence of the Endophytic Bacterium Bacillus thuringiensis Strain KB1, a Potential Biocontrol Agent against Phytopathogens.

    Science.gov (United States)

    Jeong, Haeyoung; Jo, Sung Hee; Hong, Chi Eun; Park, Jeong Mee

    2016-04-21

    ITALIC! Bacillus thuringiensisis the most widely known microbial pesticide used in agricultural applications. Herein, we report a draft genome sequence of the endophytic bacterium ITALIC! Bacillus thuringiensisstrain KB1, which exhibits antagonism against phytopathogens.

  13. Photoproduction of hydrogen by a non-sulphur bacterium isolated from root zones of water fern Azolla pinnata

    Energy Technology Data Exchange (ETDEWEB)

    Singh, S.P.; Srivastava, S.C.; Pandey, K.D. (Banaras Hindu Univ., Varanasi (IN). Centre of Advanced Study in Botany)

    1990-01-01

    A photosynthetic bacterium Rhodopseudomonas sp. BHU strain 1 was isolated from the root zone of water fern Azolla pinnata. The bacterium was found to produce hydrogen with potato starch under phototrophic conditions. The immobilized bacterial cells showed sustained hydrogen production with a more than 4-fold difference over free cell suspensions. The data have been discussed in the light of possible utilization of relatively cheaper raw materials by non-sulphur bacteria to evolve hydrogen. (author).

  14. Features of a Clostridium, strain CV-AA1, an obligatory anaerobic bacterium producing acetic acid from methanol.

    Science.gov (United States)

    Adamse, A D; Velzeboer, C T

    1982-01-01

    Isolation and characterization of a new, obligatory, anaerobic, methylotrophic, homoacetogenic bacterium is described. This bacterium is a mesophilic, motile, slightly curved rod that demonstrated a negative Gram reaction, formed spherical, (sub)terminal spores and performed a homoacetic fermentation with methanol, a CO2-2H2-gas mixture, glucose or fructose, respectively, as the substrate. The methanol fermentation proceeded only when a suitable amount of NaHCO3 was available in the nutrient solution supplied.

  15. Alcanivorax dieselolei, an alkane-degrading bacterium associated with the mucus of the zoanthid Palythoa caribaeorum (Cnidaria, Anthozoa

    Directory of Open Access Journals (Sweden)

    FF. Campos

    Full Text Available Analyses of 16S rDNA genes were used to identify the microbiota isolated from the mucus of the zoanthid Palythoa caribaeorum at Porto de Galinhas on the coast of Pernambuco State, Brazil. This study is important as the first report of this association, because of the potential biotechnological applications of the bacterium Alcanivorax dieselolei, and as evidence for the presence of a hydrocarbon degrading bacterium in a reef ecosystem such as Porto de Galinhas.

  16. A Literature Review of the Bacterium Klebsiella spp.: Grays Harbor and Chehalis River Improvements to Navigation Environmental Studies,

    Science.gov (United States)

    1981-04-01

    Taxonomy of Klebsiella pneumoniae isolated from pulp/paper mill wastewater . Environmental Protection Agency, 660/2-75-024. Knittel, M. D. 1975...AD-AI 263 CORPS OF ENGINEERS SEATTLE WASH SEATTLE DISTRICT F/6 13/2 L.ITERATURE REVIEW OF THE BACTERIUM KLEBSIELLA SPP.I BRAYS HAR--ETC(Ul...UNCLASSIFIED N Ehhmmmhhh TA GRAYS HARBOR AND CHEHALIS RIVER IMPROVEMENTS TO NAVIGATIO ENVIRONMENTAL STUDIES A LITERATURE REVIEW OF THE BACTERIUM KLEBSIELLA SPP

  17. Anaerobic, Nitrate-Dependent Oxidation of U(IV) Oxide Minerals by the Chemolithoautotrophic Bacterium Thiobacillus denitrificans

    Energy Technology Data Exchange (ETDEWEB)

    Beller, H R

    2004-06-25

    Under anaerobic conditions and at circumneutral pH, cells of the widely-distributed, obligate chemolithoautotrophic bacterium Thiobacillus denitrificans oxidatively dissolved synthetic and biogenic U(IV) oxides (uraninite) in nitrate-dependent fashion: U(IV) oxidation required the presence of nitrate and was strongly correlated to nitrate consumption. This is the first report of anaerobic U(IV) oxidation by an autotrophic bacterium.

  18. Effect of arsenite-oxidizing bacterium B. laterosporus on arsenite toxicity and arsenic translocation in rice seedlings.

    Science.gov (United States)

    Yang, Gui-Di; Xie, Wan-Ying; Zhu, Xi; Huang, Yi; Yang, Xiao-Jun; Qiu, Zong-Qing; Lv, Zhen-Mao; Wang, Wen-Na; Lin, Wen-Xiong

    2015-10-01

    Arsenite [As (III)] oxidation can be accelerated by bacterial catalysis, but the effects of the accelerated oxidation on arsenic toxicity and translocation in rice plants are poorly understood. Herein we investigated how an arsenite-oxidizing bacterium, namely Brevibacillus laterosporus, influences As (III) toxicity and translocation in rice plants. Rice seedlings of four cultivars, namely Guangyou Ming 118 (GM), Teyou Hang II (TH), Shanyou 63 (SY) and Minghui 63 (MH), inoculated with or without the bacterium were grown hydroponically with As (III) to investigate its effects on arsenic toxicity and translocation in the plants. Percentages of As (III) oxidation in the solutions with the bacterium (100%) were all significantly higher than those without (30-72%). The addition of the bacterium significantly decreased As (III) concentrations in SY root, GM root and shoot, while increased the As (III) concentrations in the shoot of SY, MH and TH and in the root of MH. Furthermore, the As (III) concentrations in the root and shoot of SY were both the lowest among the treatments with the bacterium. On the other hand, its addition significantly alleviated the As (III) toxicity on four rice cultivars. Among the treatments amended with B. laterosporus, the bacterium showed the best remediation on SY seedlings, with respect to the subdued As (III) toxicity and decreased As (III) concentration in its roots. These results indicated that As (III) oxidation accelerated by B. laterosporus could be an effective method to alleviate As (III) toxicity on rice seedlings.

  19. Role of the cell envelope stress regulators BaeR and CpxR in control of RND-type multidrug efflux pumps and transcriptional cross talk with exopolysaccharide synthesis in Erwinia amylovora.

    Science.gov (United States)

    Pletzer, Daniel; Stahl, Antje; Oja, Anna Elisabeth; Weingart, Helge

    2015-08-01

    The purpose of this study was to identify the role of the cell envelope stress-sensing systems BaeSR and CpxARP in regulation of multidrug efflux and exopolysaccharide synthesis in Erwinia amylovora. We have previously reported that BaeR activates transcription of the RND-type efflux pumps AcrD and MdtABC. In this study, we found that a cpxR-deficient mutant was highly susceptible to β-lactams, aminoglycosides and lincomycin, whereas a baeR mutant showed no change in antimicrobial sensitivity. However, overexpression of BaeR in a mutant lacking the major RND pump AcrB increased resistance of E. amylovora to several compounds that are not substrates of AcrD or MdtABC. Furthermore, we observed that overexpression of BaeR significantly increased amylovoran production. Moreover, the expression of RND-type efflux pumps was changed in regulatory mutants of exopolysaccharide production. Our data suggest that BaeSR and CpxARP regulate additional mechanisms, beside efflux, which are responsible for antimicrobial resistance of E. amylovora.

  20. EAsdiA Gene Clone and Detection to Erwinia amylovora as a New Target%EAsdiA基因的克隆和作为新靶标对梨火疫病菌的检测

    Institute of Scientific and Technical Information of China (English)

    高岩; 宋俊贤; 张蕾; 姜英华; 胡白石; 刘凤权; 许志刚

    2007-01-01

    基因sdiA属于与群体效应相关的LuxR家族,目前已证实存在于Escherichia coli和Salmonella typhimurium基因组中.本研究从梨火疫病菌(Erwinia amylovora)中克隆到了一个sdiA的同源基因,命名为EAsdiA(GenBank登录号:AY864839),该基因与E.coli和S.typhimurium的sdiA基因在氨基酸水平上分别有45.42%和43.33%的同源性,与其它细菌的luxR同源基因的同源性更低.根据梨火疫病菌EAsdiA和其它病原细菌的luxR基因的序列比对设计了1对特异引物F-EAluxR和R-EAluxR,能够特异地检测梨火疫病菌,检测灵敏度为10个菌体,在含有梨组织浸出液中可检测到102个菌体.本研究首次从梨火疫病菌中克隆sdiA基因并作为新靶标应用于该病菌分子检测.

  1. Mini-Tn10转座子构建欧文氏杆菌突变体的研究%Construction of Mutant of Erwinia carotovor by Transposon Mini-Tn10 Insertion

    Institute of Scientific and Technical Information of China (English)

    石岩; 刘正初; 徐君飞; 张居作; 李炫

    2010-01-01

    构建细菌突变体是发现新基因、分析基因功能、了解某些生物机理的重要途径.本研究通过转座子Mini-Tn10对迄今报道草本纤维提取效率最高的菌株胡萝卜软腐欧文氏杆菌(Erwinia carotovor)变异菌株CXJZU120进行随机突变,获得5 523个突变体.采用非纤维素降解实效法和水解圈法等功能性鉴定,筛选出3个非纤维素降解活性降低或丧失的突变体,为深入研究非纤维素降解机理,寻找与非纤维素降解相关基因并构建新一代高效菌株奠定了基础.

  2. High Prevalence of Antibodies against the Bacterium Treponema pallidum in Senegalese Guinea Baboons (Papio papio).

    Science.gov (United States)

    Knauf, Sascha; Barnett, Ulrike; Maciej, Peter; Klapproth, Matthias; Ndao, Ibrahima; Frischmann, Sieghard; Fischer, Julia; Zinner, Dietmar; Liu, Hsi

    2015-01-01

    The bacterium Treponema pallidum is known to cause syphilis (ssp. pallidum), yaws (ssp. pertenue), and endemic syphilis (ssp. endemicum) in humans. Nonhuman primates have also been reported to be infected with the bacterium with equally versatile clinical manifestations, from severe skin ulcerations to asymptomatic. At present all simian strains are closely related to human yaws-causing strains, an important consideration for yaws eradication. We tested clinically healthy Guinea baboons (Papio papio) at Parc National Niokolo Koba in south eastern Senegal for the presence of anti-T. pallidum antibodies. Since T. pallidum infection in this species was identified 50 years ago, and there has been no attempt to treat non-human primates for infection, it was hypothesized that a large number of West African baboons are still infected with simian strains of the yaws-bacterium. All animals were without clinical signs of treponematoses, but 18 of 20 (90%) baboons tested positive for antibodies against T. pallidum based on treponemal tests. Yet, Guinea baboons seem to develop no clinical symptoms, though it must be assumed that infection is chronic or comparable to the latent stage in human yaws infection. The non-active character is supported by the low anti-T. pallidum serum titers in Guinea baboons (median = 1:2,560) versus serum titers that are found in genital-ulcerated olive baboons with active infection in Tanzania (range of medians among the groups of initial, moderate, and severe infected animals = 1:15,360 to 1:2.097e+7). Our findings provide evidence for simian infection with T. pallidum in wild Senegalese baboons. Potentially, Guinea baboons in West Africa serve as a natural reservoir for human infection, as the West African simian strain has been shown to cause sustainable yaws infection when inoculated into humans. The present study pinpoints an area where further research is needed to support the currently on-going second WHO led yaws eradication campaign with

  3. Cloning and characterization of nif structural and regulatory genes in the purple sulfur bacterium, Halorhodospira halophila.

    Science.gov (United States)

    Tsuihiji, Hisayoshi; Yamazaki, Yoichi; Kamikubo, Hironari; Imamoto, Yasushi; Kataoka, Mikio

    2006-03-01

    Halorhodospira halophila is a halophilic photosynthetic bacterium classified as a purple sulfur bacterium. We found that H. halophila generates hydrogen gas during photoautotrophic growth as a byproduct of a nitrogenase reaction. In order to consider the applied possibilities of this photobiological hydrogen generation, we cloned and characterized the structural and regulatory genes encoding the nitrogenase, nifH, nifD and nifA, from H. halophila. This is the first description of the nif genes for a purple sulfur bacterium. The amino-acid sequences of NifH and NifD indicated that these proteins are an Fe protein and a part of a MoFe protein, respectively. The important residues are conserved completely. The sequence upstream from the nifH region and sequence similarities of nifH and nifD with those of the other organisms suggest that the regulatory system might be a NifL-NifA system; however, H. halophila lacks nifL. The amino-acid sequence of H. halophila NifA is closer to that of the NifA of the NifL-NifA system than to that of NifA without NifL. H. halophila NifA does not conserve either the residue that interacts with NifL or the important residues involved in NifL-independent regulation. These results suggest the existence of yet another regulatory system, and that the development of functional systems and their molecular counterparts are not necessarily correlated throughout evolution. All of these Nif proteins of H. halophila possess an excess of acidic residues, which acts as a salt-resistant mechanism.

  4. Whole genome shotgun sequence of Bacillus amyloliquefaciens TF28, a biocontrol entophytic bacterium.

    Science.gov (United States)

    Zhang, Shumei; Jiang, Wei; Li, Jing; Meng, Liqiang; Cao, Xu; Hu, Jihua; Liu, Yushuai; Chen, Jingyu; Sha, Changqing

    2016-01-01

    Bacillus amyloliquefaciens TF28 is a biocontrol endophytic bacterium that is capable of inhibition of a broad range of plant pathogenic fungi. The strain has the potential to be developed into a biocontrol agent for use in agriculture. Here we report the whole-genome shotgun sequence of the strain. The genome size of B. amyloliquefaciens TF28 is 3,987,635 bp which consists of 3754 protein-coding genes, 65 tandem repeat sequences, 47 minisatellite DNA, 2 microsatellite DNA, 63 tRNA, 7rRNA, 6 sRNA, 3 prophage and CRISPR domains.

  5. Asticcacaulis benevestitus sp. nov., a psychrotolerant, dimorphic, prosthecate bacterium from tundra wetland soil.

    OpenAIRE

    Vasilyeva, Lina V; Omelchenko, Marina V.; Berestovskaya, Yulia Y; Lysenko, Anatolii M; Abraham, Wolf-Rainer; Dedysh, Svetlana N.; Zavarzin, George A

    2006-01-01

    A Gram-negative, aerobic, heterotrophic, non-pigmented, dimorphic prosthecate bacterium was isolated from tundra wetland soil and designated strain Z-0023(T). Cells of this strain had a dimorphic life cycle and developed a non-adhesive stalk at a site not coincident with the centre of the cell pole, a characteristic typical of representatives of the genus Asticcacaulis. A highly distinctive feature of cells of strain Z-0023(T) was the presence of a conical, bell-shaped sheath when grown at lo...

  6. Exoelectrogenic bacterium phylogenetically related to Citrobacter freundii, isolated from anodic biofilm of a microbial fuel cell.

    Science.gov (United States)

    Huang, Jianjian; Zhu, Nengwu; Cao, Yanlan; Peng, Yue; Wu, Pingxiao; Dong, Wenhao

    2015-02-01

    An electrogenic bacterium, named Citrobacter freundii Z7, was isolated from the anodic biofilm of microbial fuel cell (MFC) inoculated with aerobic sewage sludge. Cyclic voltammetry (CV) analysis exhibited that the strain Z7 had relatively high electrochemical activity. When the strain Z7 was inoculated into MFC, the maximum power density can reach 204.5 mW/m(2) using citrate as electron donor. Series of substrates including glucose, glycerol, lactose, sucrose, and rhammose could be utilized to generate power. CV tests and the addition of anode solution as well as AQDS experiments indicated that the strain Z7 might transfer electrons indirectly via secreted mediators.

  7. A high-performance metal-free hydrogen-evolution reaction electrocatalyst from bacterium derived carbon

    OpenAIRE

    2015-01-01

    We report a sustainable approach to obtain carbon materials with nitrogen and phosphorus dual functionalities from a common bacterium strain (S. aureus) as a highly efficient hydrogen-evolution reaction (HER) catalyst. With mesoporous structure introduced by ZnCl2 salt and cathodic activation, it demonstrates an onset overpotential as low as 76 mV, a Tafel slope of 58.4 mV dec(-1) and a large normalized exchange current density of 1.72 x 10(-2) mA cm(-2), which are comparable to those of hith...

  8. Cloning, sequencing, and sequence analysis of two novel plasmids from the thermophilic anaerobic bacterium Anaerocellum thermophilum

    DEFF Research Database (Denmark)

    Clausen, Anders; Mikkelsen, Marie Just; Schrøder, I.

    2004-01-01

    The nucleotide sequence of two novel plasmids isolated from the extreme thermophilic anaerobic bacterium Anaerocellum thermophilum DSM6725 (A. thermophilum), growing optimally at 70degreesC, has been determined. pBAS2 was found to be a 3653 bp plasmid with a GC content of 43%, and the sequence...... was found, but no single stranded intermediates, characteristic of rolling circle replication, were found on Southern blots. The larger plasmid, pBAL, was found to be a 8294 bp plasmid with a GC content of 39%. It revealed 17 ORFs, of which three showed similarity at the amino acid (aa) level to known...

  9. Co-metabolism of DDT by the newly isolated bacterium, Pseudoxanthomonas sp. wax

    OpenAIRE

    2010-01-01

    Microbial degradation of 1,1,1-trichloro-2,2-bis(p-chlorophenyl)ethane (DDT) is the most promising way to clean up DDT residues found in the environment. In this paper, a bacterium designated as wax, which was capable of co-metabolizing DDT with other carbon sources, was isolated from a long-term DDT-contaminated soil sample by an enrichment culture technique. The new isolate was identified as a member of the Pseudoxanthomonas sp., based on its morphological, physiological and biochemical pro...

  10. Uncoupling effect of fatty acids in halo- and alkalotolerant bacterium Bacillus pseudofirmus FTU.

    Science.gov (United States)

    Popova, I V; Bodrova, M E; Mokhova, E N; Muntyan, M S

    2004-10-01

    Natural uncouplers of oxidative phosphorylation, long-chain non-esterified fatty acids, cause uncoupling in the alkalo- and halotolerant bacterium Bacillus pseudofirmus FTU. The uncoupling effect in the bacterial cells was manifested as decrease of membrane potential and increase of respiratory activity. The membrane potential decrease was detected only in bacterial cells exhausted by their endogenous substrates. In proteoliposomes containing reconstituted bacterial cytochrome c oxidase, fatty acids caused a "mild" uncoupling effect by reducing membrane potential only at low rate of membrane potential generation. "Free respiration" induced by the "mild" uncouplers, the fatty acids, can be considered as possible mechanism responsible for adaptation of the bacteria to a constantly changed environment.

  11. Response to Comments on "A Bacterium That Can Grow Using Arsenic Instead of Phosphorus"

    Energy Technology Data Exchange (ETDEWEB)

    Wolfe-Simon, F; Blum, J S; Kulp, T R; Gordon, G W; Hoeft, S E; Pett-Ridge, J; Stolz, J F; Webb, S M; Weber, P K; Davies, P W; Anbar, A D; Oremland, R S

    2011-03-07

    Concerns have been raised about our recent study describing a bacterium that can grow using arsenic (As) instead of phosphorus (P). Our data suggested that As could act as a substitute for P in major biomolecules in this organism. Although the issues raised are of investigative interest, we contend that they do not invalidate our conclusions. We argue that while no single line of evidence we presented was sufficient to support our interpretation of the data, taken as an entire dataset we find no plausible alternative to our conclusions. Here we reply to the critiques and provide additional arguments supporting the assessment of the data we reported.

  12. Dynamic detection of a single bacterium: nonlinear rotation rate shifts of driven magnetic microsphere stages

    CERN Document Server

    McNaughton, B H; Kopelman, R; Agayan, Rodney R.; Kopelman, Raoul; Naughton, Brandon H. Mc

    2006-01-01

    We report on a new technique which was used to detect single Escherichia coli that is based on the changes in the nonlinear rotation of a magnetic microsphere driven by an external magnetic field. The presence of one Escherichia Coli bacterium on the surface of a 2.0 micron magnetic microsphere caused an easily measurable change in the drag of the system and, therefore, in the nonlinear rotation rate. The straight-forward measurement uses standard microscopy techniques and the observed average shift in the nonlinear rotation rate changed by a factor of ~3.8.

  13. Isolation and identification of a novel alginate-degrading bacterium, Ochrobactrum sp.

    Directory of Open Access Journals (Sweden)

    Xiao-wei Zhao

    2008-03-01

    Full Text Available An alginate-degrading bacterium, identified as Ochrobactrum sp. on the basis of 16S rDNA gene sequencing, was isolated from brown algal samples collected from the waters in close vicinity to the Dongtou Isles in the East China Sea. The strain, designated WZUH09-1, is a short rod, gram-negative, obligatory aerobic, grows under the following conditions: 5-40oC, pH 3-9, and 0-2 times of the seawater concentration, and is able to depolymerize alginates with higher enzyme activity than that of others reported so far.

  14. FACTORS LIMITING BACTERIAL GROWTH : III. CELL SIZE AND "PHYSIOLOGIC YOUTH" IN BACTERIUM COLI CULTURES.

    Science.gov (United States)

    Hershey, A D; Bronfenbrenner, J

    1938-07-20

    1. Measurements of the rate of oxygen uptake per cell in transplants of Bacterium coli from cultures of this organism in different phases of growth have given results in essential agreement with the observations of others. 2. Correlations of viable count, centrifugable nitrogen, and turbidity, with oxygen consumption, indicate that the increased metabolism during the early portion of the growth period is quantitatively referable to increased average size of cells. 3. Indirect evidence has suggested that the initial rate of growth of transplants is not related to the phase of growth of the parent culture.

  15. Complete genome sequence of Enterobacter cloacae GGT036: a furfural tolerant soil bacterium.

    Science.gov (United States)

    Gong, Gyeongtaek; Um, Youngsoon; Park, Tai Hyun; Woo, Han Min

    2015-01-10

    Enterobacter cloacae is a facultative anaerobic bacterium to be an important cause of nosocomial infection. However, the isolated E. cloacae GGT036 showed higher furfural-tolerant cellular growth, compared to industrial relevant strains such as Escherichia coli and Corynebacterium glutamicum. Here, we report the complete genome sequence of E. cloacae GGT036 isolated from Mt. Gwanak, Seoul, Republic of Korea. The genomic DNA sequence of E. cloacae GGT036 will provide valuable genetic resources for engineering of industrially relevant strains being tolerant to cellular inhibitors present in lignocellulosic hydrolysates.

  16. Aggregation of the rhizospheric bacterium Azospirillum brasilense in response to oxygen

    Science.gov (United States)

    Abdoun, Hamid; McMillan, Mary; Pereg, Lily

    2016-04-01

    Azospirillum brasilense spp. have ecological, scientific and agricultural importance. As model plant growth promoting rhizobacteria they interact with a large variety of plants, including important food and cash crops. Azospirillum strains are known for their production of plant growth hormones that enhance root systems and for their ability to fix nitrogen. Azospirillum cells transform in response to environmental cues. The production of exopolysaccharides and cell aggregation during cellular transformation are important steps in the attachment of Azospirillum to roots. We investigate signals that induce cellular transformation and aggregation in the Azospirillum and report on the importance of oxygen to the process of aggregation in this rhizospheric bacterium.

  17. A bacterium that can grow by using arsenic instead of phosphorus

    Energy Technology Data Exchange (ETDEWEB)

    Wolfe-Simon, F; Blum, J S; Kulp, T R; Gordon, G W; Hoeft, S E; Pett-Ridge, J; Stolz, J F; Webb, S M; Weber, P K; Davies, P W; Anbar, A D; Oremland, R S

    2010-11-01

    Life is mostly composed of the elements carbon, hydrogen, nitrogen, oxygen, sulfur and phosphorus. Although these six elements make up nucleic acids, proteins and lipids and thus the bulk of living matter, it is theoretically possible that some other elements in the periodic table could serve the same functions. Here we describe a bacterium, strain GFAJ-1 of the Halomonadaceae, isolated from Mono Lake, CA, which substitutes arsenic for phosphorus to sustain its growth. Our data show evidence for arsenate in macromolecules that normally contain phosphate, most notably nucleic acids and proteins. Exchange of one of the major bio-elements may have profound evolutionary and geochemical significance.

  18. A bacterium that can grow by using arsenic instead of phosphorus.

    Science.gov (United States)

    Wolfe-Simon, Felisa; Switzer Blum, Jodi; Kulp, Thomas R; Gordon, Gwyneth W; Hoeft, Shelley E; Pett-Ridge, Jennifer; Stolz, John F; Webb, Samuel M; Weber, Peter K; Davies, Paul C W; Anbar, Ariel D; Oremland, Ronald S

    2011-06-03

    Life is mostly composed of the elements carbon, hydrogen, nitrogen, oxygen, sulfur, and phosphorus. Although these six elements make up nucleic acids, proteins, and lipids and thus the bulk of living matter, it is theoretically possible that some other elements in the periodic table could serve the same functions. Here, we describe a bacterium, strain GFAJ-1 of the Halomonadaceae, isolated from Mono Lake, California, that is able to substitute arsenic for phosphorus to sustain its growth. Our data show evidence for arsenate in macromolecules that normally contain phosphate, most notably nucleic acids and proteins. Exchange of one of the major bio-elements may have profound evolutionary and geochemical importance.

  19. Draft Genome Sequence of the Endophytic Strain Rhodococcus kyotonensis KB10, a Potential Biodegrading and Antibacterial Bacterium Isolated from Arabidopsis thaliana

    Science.gov (United States)

    Hong, Chi Eun; Jo, Sung Hee

    2016-01-01

    Rhodococcus kyotonensis KB10 is an endophytic bacterium isolated from Arabidopsis thaliana. The organism showed mild antibacterial activity against the phytopathogen Pseudomonas syringae pv. tomato DC3000. This study reports the genome sequence of R. kyotonensis KB10. This bacterium contains an ectoine biosynthesis gene cluster and has the potential to degrade nitroaromatic compounds. The identified bacterium may be a suitable biocontrol agent and degrader of environmental pollutants. PMID:27389269

  20. Non-specific immune response of bullfrog Rana catesbeiana to intraperitoneal injection of bacterium Aeromonas hydrophila

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    Non-specific immune response of bullfrog Rana catesbeiana to pathogenic Aeromonas hydrophila was studied to 60 individuals in two groups. Each bullfrog in bacterium-injected group was injected intraperitoneally (i.p.) with 0.2 ml bacterial suspension at a density of 5.2 × 106 CFU/ml, while each one in control group injected i.p. with 0.2 ml sterile saline solution (0.85%, w/v). Three bullfrogs in both groups were sampled at 0, 1, 3, 7, 11, 15 and 20 days post-injection (dpi) for the evaluation of non-specific immune parameters. It was observed that intraperitoneal injection of A. hydrophila significantly increased the number of leucocytes and that of NBT-positive cells in peripheral blood. Significant increases in serum bactericidal activity and serum acid phosphatase activity were also observed in the bacterium-injected frogs when compared with those in the control group. However, a significant reduction was detected in vitro in phagocytosis activity of peripheral blood phagocytes. No significant difference in changes in the number of peripheral erythrocytes, serum superoxide dismutase (SOD) activity, and lysozyme activity was detected between the two groups. It is suggested that bullfrogs may produce a series of non-specific immune reactions in response to the A. hydrophila infection.