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Sample records for enterobacterial phytopathogen erwinia

  1. The Phytopathogen Dickeya dadantii (Erwinia chrysanthemi 3937) Is a Pathogen of the Pea Aphid†

    OpenAIRE

    Grenier, Anne-Marie; Duport, Gabrielle; Pagès, Sylvie; Condemine, Guy; Rahbé, Yvan

    2006-01-01

    Dickeya dadantii (Erwinia chrysanthemi) is a phytopathogenic bacterium causing soft rot diseases on many crops. The sequencing of its genome identified four genes encoding homologues of the Cyt family of insecticidal toxins from Bacillus thuringiensis, which are not present in the close relative Pectobacterium carotovorum subsp. atrosepticum. The pathogenicity of D. dadantii was tested on the pea aphid Acyrthosiphon pisum, and the bacterium was shown to be highly virulent for this insect, eit...

  2. The phytopathogen Dickeya dadantii (Erwinia chrysanthemi 3937) is a pathogen of the pea aphid.

    Science.gov (United States)

    Grenier, Anne-Marie; Duport, Gabrielle; Pagès, Sylvie; Condemine, Guy; Rahbé, Yvan

    2006-03-01

    Dickeya dadantii (Erwinia chrysanthemi) is a phytopathogenic bacterium causing soft rot diseases on many crops. The sequencing of its genome identified four genes encoding homologues of the Cyt family of insecticidal toxins from Bacillus thuringiensis, which are not present in the close relative Pectobacterium carotovorum subsp. atrosepticum. The pathogenicity of D. dadantii was tested on the pea aphid Acyrthosiphon pisum, and the bacterium was shown to be highly virulent for this insect, either by septic injury or by oral infection. The lethal inoculum dose was calculated to be as low as 10 ingested bacterial cells. A D. dadantii mutant with the four cytotoxin genes deleted showed a reduced per os virulence for A. pisum, highlighting the potential role of at least one of these genes in pathogenicity. Since only one bacterial pathogen of aphids has been previously described (Erwinia aphidicola), other species from the same bacterial group were tested. The pathogenic trait for aphids was shown to be widespread, albeit variable, within the phytopathogens, with no link to phylogenetic positioning in the Enterobacteriaceae. Previously characterized gut symbionts from thrips (Erwinia/Pantoea group) were also highly pathogenic to the aphid, whereas the potent entomopathogen Photorhabdus luminescens was not. D. dadantii is not a generalist insect pathogen, since it has low pathogenicity for three other insect species (Drosophila melanogaster, Sitophilus oryzae, and Spodoptera littoralis). D. dadantii was one of the most virulent aphid pathogens in our screening, and it was active on most aphid instars, except for the first one, probably due to anatomical filtering. The observed difference in virulence toward apterous and winged aphids may have an ecological impact, and this deserves specific attention in future research.

  3. The structure and significance of enterobacterial common antigen (ECA

    Directory of Open Access Journals (Sweden)

    Tomasz Kasper Goździewicz

    2015-09-01

    Full Text Available The enterobacterial common antigen (ECA is a carbohydrate-derived cell surface antigen present in all Gram-negative bacteria belonging to Enterobacteriaceae family. Biosynthetic pathways shared by ECA and LPS (endotoxin suggest close connections between these antigens. ECA occurs in three different forms: a phosphatidyl-linked linear polysaccharide anchored on the cell surface (ECAPG, a cyclic form built of 4-6 repeating units localized in the periplasm (ECACYC and as a linear polysaccharide covalently linked to LPS core oligosaccharide (ECALPS. Regardless of ECA form, poly- and oligosaccharides of ECA consist of the biological trisaccharide repeating units: →3-α-d-Fucp4NAc-(1→4-β-d-ManpNAcA-(1→4-α-d-GlcpNAc-(1→, where Fucp4NAc refers to 4-acetamido-2,4-dideoxygalactose, ManpNAcA to N-acetyl-mannosaminuronic acid and GlcpNAc to N-acetylglucosamine. ECAPG and ECALPS consisting of one unit with Fucp4NAc as a terminal sugar were also identified. The number of the studies shows its occurrence in all members of enteric bacteria with a few exceptions such as Erwinia chrysanthemi. The presence of ECA was also shown for such genera as Plesiomonas [4] and Yersinia [36], previously belonging to the Vibrionaceae and Pasteurellaceae families, respectively. It was one of the reasons to include these two taxa in the Enterobacteriaceae family. The function of ECA is not fully understood, but it was reported that its occurrence is important in resistance of bacterial cells to environmental conditions, such as bile salts in the human digestive tract. The immunogenicity of ECA seems very interesting in the fact that only sparse rough Gram-negative strains, such as Shigella sonnei phase II, Escherichia coli R1, R2, R4, K-12, and Yersinia enterocolitica O:3 are able to induce the production of specific anti-ECA antibodies. It is the effect of the ECALPS, and the evidence for the existence of such covalent linkage was provided by structural analysis of S

  4. Growth of bacterial phytopathogens in animal manures.

    Science.gov (United States)

    Sledz, Wojciech; Zoledowska, Sabina; Motyka, Agata; Kadziński, Leszek; Banecki, Bogdan

    2017-01-01

    Animal manures are routinely applied to agricultural lands to improve crop yield, but the possibility to spread bacterial phytopathogens through field fertilization has not been considered yet. We monitored 49 cattle, horse, swine, sheep or chicken manure samples collected in 14 Polish voivodeships for the most important plant pathogenic bacteria - Ralstonia solanacearum (Rsol), Xanthomonas campestris pv. campestris (Xcc), Pectobacterium carotovorum subsp. carotovorum (Pcc), Pectobacterium atrosepticum (Pba), Erwinia amylovora (Eam), Clavibacter michiganensis subsp. sepedonicus (Cms) and Dickeya sp. (Dsp). All of the tested animal fertilizers were free of these pathogens. Subsequently, the growth dynamics of Pba, Pcc, Rsol, and Xcc in cattle, horse, swine, sheep and chicken manures sterilized either by autoclaving or filtration was evaluated. The investigated phytopathogens did not exhibit any growth in the poultry manure. However, the manure filtrates originating from other animals were suitable for microbial growth, which resulted in the optical density change of 0.03-0.22 reached within 26 h (48 h Rsol, 120 h Xcc), depending on bacterial species and the manure source. Pcc and Pba multiplied most efficiently in the cattle manure filtrate. These bacteria grew faster than Rsol and Xcc in all the tested manure samples, both the filtrates and the autoclaved semi-solid ones. Though the growth dynamics of investigated strains in different animal fertilizers was unequal, all of the tested bacterial plant pathogens were proven to use cattle, horse, swine and sheep manures as the sources of nutrients. These findings may contribute to further research on the alternative routes of spread of bacterial phytopathogens, especially because of the fact that the control of pectionolytic bacteria is only based on preventive methods.

  5. [Enterobacterial antigen in human peripheral blood lymphocytes].

    Science.gov (United States)

    Faure-Fontenla, M A; García-Tamayo, F

    1989-11-01

    The following study has as prior history the research reports which have shown the existence of an antigenic tissue deposit in gram-negative enterobacteria. The antigens of the enterobacteria have also been found in the lymphocytic membranes and cytoplasm. Since intestinal lymphoid tissue cells can recirculate by means of the thoracic duct to the peripheral venous system, it was proposed that the circulating lymphocytes in healthy people could also contain small amounts of a common enterobacterial antigen. The study was carried out in 15 human venous blood samples, of which the lymphocytic population was separated to later be used in the preparation of 15 alcohol soluble extracts. This material was used for inhibiting the immuno-hemolysis assay in three occasions in order to show the presence of antigens shared by different enterobacterias, using as reference a fraction separated from the LPS of Escherichia coli 08. The results showed that the human lymphocytes also had antigenic determinants common to gram-negative bacteria.

  6. 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.

  7. 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

  8. Enterobacterial involvement in the pathogenesis of secondary ankylosing spondylitis.

    Science.gov (United States)

    van Bohemen, C G; Weterings, E; Goei The, H S; Grumet, F C; Zanen, H C

    1988-01-01

    Ankylosing spondylitis (AS) is closely associated with the histocompatibility antigen HLA-B27. Pathogenesis of AS is thought to involve interactions between B27 and certain enterobacterial antigens. However, this is uncertain and contested by some. The present paper argues that the presence of statistically raised specific serum IgA to a common enterobacterial heat modifiable major outer membrane protein (h-momp; Mr 35,000) in active AS (N = 25; IgA = 1485 +/- 20) in comparison to controls, most notably hospital patients without known arthropathies or gastrointestinal disease (N = 12; IgA = 548 +/- 59), supports an inductive contribution of enterobacterial antigens to the pathogenesis of secondary AS. Serum IgG and IgM did not statistically differ. Raised specific serum IgA to h-momp might indicate enterobacterial antigenic stimulation from the gastrointestinal tract. It does not necessarily imply direct involvement in the pathogenesis of primary AS. H-momp appears to be a convenient tool for serological studies of AS and at present is likely to be more suitable than other bacterial antigens, notably those with B27-like epitopes. Namely, the confirmed presence in AS of enterobacteria with freely accessible B27-like antigenic epitopes on their cell surface might induce unusual tolerance to these organisms in B27 positive hosts, thus causing chronic inflammation, initially sacroiliitis (and spondylitis) due to the proximity of presacral and para-aortic colon draining lymph nodes, later becoming more generalized (for reasons unclear) to include other lesions (e.g. peripheral arthritis, uveitis, enthesopathies). Thus, antibodies to B27-like antigenic epitopes need not be detectable or may be absent. Also, cellular immune responsiveness to these antigens might be involved.

  9. Veredeling brengt inzicht in Erwinia-resistentie

    NARCIS (Netherlands)

    Snijder, R.; Paasen, van M.Q.P.; Tuyl, van J.M.

    2000-01-01

    Plant Research International, het voormalige CPRO-DLO, doet in opdracht van zes bedrijven onderzoek naar resistentiebronnen binnen Zantedeschia. Kort verslag van de resistentietoets voor Erwinia-rot en de eerste resultaten binnen het sortiment

  10. 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.

  11. Soft rot erwiniae: from genes to genomes.

    Science.gov (United States)

    Toth, Ian K; Bell, Kenneth S; Holeva, Maria C; Birch, Paul R J

    2003-01-01

    SUMMARY The soft rot erwiniae, Erwinia carotovora ssp. atroseptica (Eca), E. carotovora ssp. carotovora (Ecc) and E. chrysanthemi (Ech) are major bacterial pathogens of potato and other crops world-wide. We currently understand much about how these bacteria attack plants and protect themselves against plant defences. However, the processes underlying the establishment of infection, differences in host range and their ability to survive when not causing disease, largely remain a mystery. This review will focus on our current knowledge of pathogenesis in these organisms and discuss how modern genomic approaches, including complete genome sequencing of Eca and Ech, may open the door to a new understanding of the potential subtlety and complexity of soft rot erwiniae and their interactions with plants. The soft rot erwiniae are members of the Enterobacteriaceae, along with other plant pathogens such as Erwinia amylovora and human pathogens such as Escherichia coli, Salmonella spp. and Yersinia spp. Although the genus name Erwinia is most often used to describe the group, an alternative genus name Pectobacterium was recently proposed for the soft rot species. Ech mainly affects crops and other plants in tropical and subtropical regions and has a wide host range that includes potato and the important model host African violet (Saintpaulia ionantha). Ecc affects crops and other plants in subtropical and temperate regions and has probably the widest host range, which also includes potato. Eca, on the other hand, has a host range limited almost exclusively to potato in temperate regions only. Disease symptoms: Soft rot erwiniae cause general tissue maceration, termed soft rot disease, through the production of plant cell wall degrading enzymes. Environmental factors such as temperature, low oxygen concentration and free water play an essential role in disease development. On potato, and possibly other plants, disease symptoms may differ, e.g. blackleg disease is associated

  12. Differentiation of Phytopathogenic agrobacterium spp.

    OpenAIRE

    Kuzmanović, Nemanja; Ivanović, Milan; Ćalić, Anđelka; Gašić, Katarina; Obradović, Aleksa

    2011-01-01

    Due to the difficulties in differentiation of phytopathogenic Agrobacterium spp. and lack of a standardized protocol, we carried out selection and evaluation of suitable methods based on the bacterial physiological, genetic and pathogenic properties. Strains of Agrobacterium tumefaciens, A. rhizogenes and A. vitis were differentiated using standard bacteriological and molecular methods. The biochemical and physiological tests confirmed authenticity of the s...

  13. Genetically pyramiding protease-inhibitor genes for dual broad-spectrum resistance against insect and phytopathogens in transgenic tobacco.

    Science.gov (United States)

    Senthilkumar, Rajendran; Cheng, Chiu-Ping; Yeh, Kai-Wun

    2010-01-01

    Protease inhibitors provide a promising means of engineering plant resistance against attack by insects and pathogens. Sporamin (trypsin inhibitor) from sweet potato and CeCPI (phytocystatin) from taro were stacked in a binary vector, using pMSPOA (a modified sporamin promoter) to drive both genes. Transgenic tobacco lines of T0 and T1 generation with varied inhibitory activity against trypsin and papain showed resistance to both insects and phytopathogens. Larvae of Helicoverpa armigera that ingested tobacco leaves either died or showed delayed growth and development relative to control larvae. Transgenic tobacco-overexpressing the stacked genes also exhibited strong resistance against bacterial soft rot disease caused by Erwinia carotovora and damping-off disease caused by Pythium aphanidermatum. Thus, stacking protease-inhibitor genes, driven by the wound and pathogen responsive pMSPOA promoter, is an effective strategy for engineering crops to resistance against insects and phytopathogens.

  14. Biochemical adaptation of phytopathogenic fungi, Sclerotium rolfsii ...

    African Journals Online (AJOL)

    Biochemical adaptation of phytopathogenic fungi, Sclerotium rolfsii, in response to temperature stress. Natthiya Buensanteai, Kanjana Thumanu, Khanistha Kooboran, Dusit Athinuwat, Sutruedee Prathuangwong ...

  15. Clinical strains of Enterobacter agglomerans (synonyms: Erwinia herbicola, Erwinia milletiae) identified by DNA-DNA-hybridization.

    Science.gov (United States)

    Lind, E; Ursing, J

    1986-08-01

    Using DNA-DNA-hybridization it could be shown that 52 of 86 clinical isolates of Enterobacter agglomerans were closely related to each other, to the type strain of the species and also to the type strains of Erwinia herbicola and Erwinia milletiae. Most of the strains investigated were of the biogroups 1 and G1 of Ewing & Fife. All strains of the genetically defined group belonged to these two biogroups; none of these isolates fermented dulcitol, and with few exceptions they were also cellobiose, lactose and sorbitol negative.

  16. Pseudomonas orientalis F9: A Potent Antagonist against Phytopathogens with Phytotoxic Effect in the Apple Flower

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    Veronika Zengerer

    2018-02-01

    Full Text Available In light of public concerns over the use of pesticides and antibiotics in plant protection and the subsequent selection for spread of resistant bacteria in the environment, it is inevitable to broaden our knowledge about viable alternatives, such as natural antagonists and their mode of action. The genus Pseudomonas is known for its metabolic versatility and genetic plasticity, encompassing pathogens as well as antagonists. We characterized strain Pseudomonas orientalis F9, an isolate from apple flowers in a Swiss orchard, and determined its antagonistic activity against several phytopathogenic bacteria, in particular Erwinia amylovora, the causal agent of fire blight. P. orientalis F9 displayed antagonistic activity against a broad suite of phytopathogenic bacteria in the in vitro tests. The promising results from this analysis led to an ex vivo assay with E. amylovora CFBP1430Rif and P. orientalis F9 infected detached apple flowers. F9 diminished the fire blight pathogen in the flowers but also revealed phytotoxic traits. The experimental results were discussed in light of the complete genome sequence of F9, which revealed the strain to carry phenazine genes. Phenazines are known to contribute to antagonistic activity of bacterial strains against soil pathogens. When tested in the cress assay with Pythium ultimum as pathogen, F9 showed results comparable to the known antagonist P. protegens CHA0.

  17. Pseudomonas orientalis F9: A Potent Antagonist against Phytopathogens with Phytotoxic Effect in the Apple Flower.

    Science.gov (United States)

    Zengerer, Veronika; Schmid, Michael; Bieri, Marco; Müller, Denise C; Remus-Emsermann, Mitja N P; Ahrens, Christian H; Pelludat, Cosima

    2018-01-01

    In light of public concerns over the use of pesticides and antibiotics in plant protection and the subsequent selection for spread of resistant bacteria in the environment, it is inevitable to broaden our knowledge about viable alternatives, such as natural antagonists and their mode of action. The genus Pseudomonas is known for its metabolic versatility and genetic plasticity, encompassing pathogens as well as antagonists. We characterized strain Pseudomonas orientalis F9, an isolate from apple flowers in a Swiss orchard, and determined its antagonistic activity against several phytopathogenic bacteria, in particular Erwinia amylovora , the causal agent of fire blight. P. orientalis F9 displayed antagonistic activity against a broad suite of phytopathogenic bacteria in the in vitro tests. The promising results from this analysis led to an ex vivo assay with E. amylovora CFBP1430 Rif and P. orientalis F9 infected detached apple flowers. F9 diminished the fire blight pathogen in the flowers but also revealed phytotoxic traits. The experimental results were discussed in light of the complete genome sequence of F9, which revealed the strain to carry phenazine genes. Phenazines are known to contribute to antagonistic activity of bacterial strains against soil pathogens. When tested in the cress assay with Pythium ultimum as pathogen, F9 showed results comparable to the known antagonist P. protegens CHA0.

  18. Distribution of phytopathogenic bacteria in infested seeds

    Science.gov (United States)

    Populations of phytopathogenic bacteria representing five host-pathogen combinations were assessed to determine if there was a mathematical relationship common across seedborne bacterial diseases. Bacterial populations were estimated from naturally-infested seeds of cowpea (Vigna unguiculata), peppe...

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

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    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.

  20. Erwinia in Zantedeschia : volgen van partijen en toetsen op tolerantie

    NARCIS (Netherlands)

    Leeuwen, van P.J.; Martin, W.S.; Dees, R.H.L.; Trompert, J.P.T.; Doorn, van J.

    2012-01-01

    De bloembollen- en de aardappelsector trekken gezamenlijk op binnen het koepel project Deltaplan Erwinia om het Erwiniaprobleem aan te pakken. Voor Zantedeschia is alleen de Erwiniabacteriesoort Pectobacterium carotovorum subsp. carotovorum (Pcc) een groot probleem. De afgelopen jaren is onderzocht

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

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    Veljko Gavrilović

    2008-01-01

    Full Text Available 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 investigation was conducted to identify the causal agent of this disease. Levan-producing, nonfluorescent bacteria were isolated from diseased samples. The isolated strains produced HR in tobacco leaves and necrosis of artificially inoculated, immature pear fruits, followed by oozing of bacterial exudate, a characterisitic of E. amylovora. Based on the results of pathogenicity tests, biochemical characteristics, ELISA test and PCR analysis, it was confirmed that the investigated strainsbelonged to E. amylovora, causing the root collar necrosis of apple trees as an atypical symptom of this bacterium in Serbia. The explanation of this symptom may be that the vegetative rootstocks were infected with E. amylovora. Therefore, the development of diagnostic protocols for detection of E. amylovora in apple rootstock is very important for health inspections of planting materials.

  2. Differentiation of Phytopathogenic Agrobacterium spp.

    Directory of Open Access Journals (Sweden)

    Nemanja Kuzmanović

    2011-01-01

    Full Text Available Due to the difficulties in differentiation of phytopathogenic Agrobacterium spp. and lack of a standardized protocol, we carried out selection and evaluation of suitable methods based on the bacterial physiological, genetic and pathogenic properties. Strains of Agrobacterium tumefaciens, A. rhizogenes and A. vitis were differentiated using standard bacteriological and molecular methods. The biochemical and physiological tests confirmed authenticity of the strains. Two duplex PCR methods were conducted with four different primer pairs. In all strains, presence of plasmid virD2 and virC pathogenicity genes was detected. Chromosomal pehA gene was determined in A. vitis strain. Pathogenicity was confirmed on carrot slices and young plants of tomato and sunflower. Strains of A. tumefaciens and A. vitis were pathogenic on all test plants, while strain of A. rhizogenes induced characteristic symptoms only on carrot slices. The tests used in this study provided reliable discrimination between the three species and confirmed their identity as tumorigenic (TiAgrobacterium tumefaciens and A. vitis, and rhizogenic (Ri A. rhizogenes.

  3. Genera of phytopathogenic fungi: GOPHY 1

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    Y. Marin-Felix

    2017-03-01

    Full Text Available Genera of Phytopathogenic Fungi (GOPHY is introduced as a new series of publications in order to provide a stable platform for the taxonomy of phytopathogenic fungi. This first paper focuses on 21 genera of phytopathogenic fungi: Bipolaris, Boeremia, Calonectria, Ceratocystis, Cladosporium, Colletotrichum, Coniella, Curvularia, Monilinia, Neofabraea, Neofusicoccum, Pilidium, Pleiochaeta, Plenodomus, Protostegia, Pseudopyricularia, Puccinia, Saccharata, Thyrostroma, Venturia and Wilsonomyces. For each genus, a morphological description and information about its pathology, distribution, hosts and disease symptoms are provided. In addition, this information is linked to primary and secondary DNA barcodes of the presently accepted species, and relevant literature. Moreover, several novelties are introduced, i.e. new genera, species and combinations, and neo-, lecto- and epitypes designated to provide a stable taxonomy. This first paper includes one new genus, 26 new species, ten new combinations, and four typifications of older names.

  4. Biochemical adaptation of phytopathogenic fungi, Sclerotium rolfsii ...

    African Journals Online (AJOL)

    nbuensanteai

    2012-10-18

    Oct 18, 2012 ... Temperature stress plays a critical influence on microbial survival and ecology. It has been reported to be associated with direct effects on microbial metabolisms, but there are very few studies in literature which have reported it in phytopathogenic fungi. In this study, we investigated the impact of two.

  5. Restriction enzyme fingerprinting of enterobacterial plasmids: a simple strategy with wide application.

    OpenAIRE

    Platt, D. J.; Chesham, J. S.; Brown, D. J.; Kraft, C. A.; Taggart, J.

    1986-01-01

    Restriction enzyme fingerprints were generated from purified plasmid DNA from 324 clinical isolates that belonged to 7 enterobacterial genera and 88 single plasmids in Escherichia coli K 12 according to the following strategy. Purified plasmid DNA was digested with PstI. The number of fragments detected in a 0.8 agarose gel was used to determine which 2 of 6 restriction enzymes including PstI was most likely to provide a fingerprint comprising sufficient fragments to ensure specificity but su...

  6. Resistance to Erwinia spp. in potato (Solanum tuberosum L.)

    NARCIS (Netherlands)

    Allefs, S.

    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

  7. Genome Sequences of 19 Novel Erwinia amylovora Bacteriophages

    Science.gov (United States)

    Esplin, Ian N. D.; Berg, Jordan A.; Sharma, Ruchira; Allen, Robert C.; Arens, Daniel K.; Ashcroft, Cody R.; Bairett, Shannon R.; Beatty, Nolan J.; Bickmore, Madeline; Bloomfield, Travis J.; Brady, T. Scott; Bybee, Rachel N.; Carter, John L.; Choi, Minsey C.; Duncan, Steven; Fajardo, Christopher P.; Foy, Brayden B.; Fuhriman, David A.; Gibby, Paul D.; Grossarth, Savannah E.; Harbaugh, Kala; Harris, Natalie; Hilton, Jared A.; Hurst, Emily; Hyde, Jonathan R.; Ingersoll, Kayleigh; Jacobson, Caitlin M.; James, Brady D.; Jarvis, Todd M.; Jaen-Anieves, Daniella; Jensen, Garrett L.; Knabe, Bradley K.; Kruger, Jared L.; Merrill, Bryan D.; Pape, Jenny A.; Payne Anderson, Ashley M.; Payne, David E.; Peck, Malia D.; Pollock, Samuel V.; Putnam, Micah J.; Ransom, Ethan K.; Ririe, Devin B.; Robinson, David M.; Rogers, Spencer L.; Russell, Kerri A.; Schoenhals, Jonathan E.; Shurtleff, Christopher A.; Simister, Austin R.; Smith, Hunter G.; Stephenson, Michael B.; Staley, Lyndsay A.; Stettler, Jason M.; Stratton, Mallorie L.; Tateoka, Olivia B.; Tatlow, P. J.; Taylor, Alexander S.; Thompson, Suzanne E.; Townsend, Michelle H.; Thurgood, Trever L.; Usher, Brittian K.; Whitley, Kiara V.; Ward, Andrew T.; Ward, Megan E. H.; Webb, Charles J.; Wienclaw, Trevor M.; Williamson, Taryn L.; Wells, Michael J.; Wright, Cole K.; Breakwell, Donald P.; Hope, Sandra

    2017-01-01

    ABSTRACT Erwinia amylovora is the causal agent of fire blight, a devastating disease affecting some plants of the Rosaceae family. We isolated bacteriophages from samples collected from infected apple and pear trees along the Wasatch Front in Utah. We announce 19 high-quality complete genome sequences of E. amylovora bacteriophages. PMID:29146842

  8. Genome Sequences of 19 Novel Erwinia amylovora Bacteriophages

    OpenAIRE

    Esplin, Ian N. D.; Berg, Jordan A.; Sharma, Ruchira; Allen, Robert C.; Arens, Daniel K.; Ashcroft, Cody R.; Bairett, Shannon R.; Beatty, Nolan J.; Bickmore, Madeline; Bloomfield, Travis J.; Brady, T. Scott; Bybee, Rachel N.; Carter, John L.; Choi, Minsey C.; Duncan, Steven

    2017-01-01

    ABSTRACT Erwinia amylovora is the causal agent of fire blight, a devastating disease affecting some plants of the Rosaceae family. We isolated bacteriophages from samples collected from infected apple and pear trees along the Wasatch Front in Utah. We announce 19 high-quality complete genome sequences of E. amylovora bacteriophages.

  9. Erwinia chrysanthemi ook bij ploffers in Dahlia boosdoener

    NARCIS (Netherlands)

    Leeuwen, van P.J.; Trompert, J.P.T.

    2006-01-01

    Sinds een aantal jaren komt bij de stekproduktie van Dahlia veel uitval voor door ploffers. Na het verhogen van de kastemperatuur vallen de knollen natrot weg. Bovendien kan verdere besmetting snel om zich heen grijpen. Onderzoek heeft aangetoond dat de bacterie Erwinia chrysanthemi de veroorzaker

  10. Lon protease modulates virulence traits in Erwinia amylovora by direct monitoring of major regulators and indirectly through the Rcs and Gac-Csr regulatory systems.

    Science.gov (United States)

    Lee, Jae Hoon; Ancona, Veronica; Zhao, Youfu

    2018-04-01

    Lon, an ATP-dependent protease in bacteria, influences diverse cellular processes by degrading damaged, misfolded and short-lived regulatory proteins. In this study, we characterized the effects of lon mutation and determined the molecular mechanisms underlying Lon-mediated virulence regulation in Erwinia amylovora, an enterobacterial pathogen of apple. Erwinia amylovora depends on the type III secretion system (T3SS) and the exopolysaccharide (EPS) amylovoran to cause disease. Our results showed that mutation of the lon gene led to the overproduction of amylovoran, increased T3SS gene expression and the non-motile phenotype. Western blot analyses showed that mutation in lon directly affected the accumulation and stability of HrpS/HrpA and RcsA. Mutation in lon also indirectly influenced the expression of flhD, hrpS and csrB through the accumulation of the RcsA/RcsB proteins, which bind to the promoter of these genes. In addition, lon expression is under the control of CsrA, possibly at both the transcriptional and post-transcriptional levels. Although mutation in csrA abolished both T3SS and amylovoran production, deletion of the lon gene in the csrA mutant only rescued amylovoran production, but not T3SS. These results suggest that CsrA might positively control both T3SS and amylovoran production partly by suppressing Lon, whereas CsrA may also play a critical role in T3SS by affecting unknown targets. © 2017 BSPP AND JOHN WILEY & SONS LTD.

  11. Massive production of butanediol during plant infection by phytopathogenic bacteria of the genera Dickeya and Pectobacterium.

    Science.gov (United States)

    Effantin, Géraldine; Rivasseau, Corinne; Gromova, Marina; Bligny, Richard; Hugouvieux-Cotte-Pattat, Nicole

    2011-11-01

    Plant pathogenic bacteria of the genera Dickeya and Pectobacterium are broad-host-range necrotrophs which cause soft-rot diseases in important crops. A metabolomic analysis, based on (13)C-NMR spectroscopy, was used to characterize the plant-bacteria interaction. Metabolic profiles revealed a decline in plant sugars and amino acids during infection and the concomitant appearance of a compound identified as 2,3-butanediol. Butanediol is the major metabolite found in macerated tissues of various host plants. It is accumulated during the symptomatic phase of the disease. Different species of Dickeya or Pectobacterium secrete high levels of butanediol during plant infection. Butanediol has been described as a signalling molecule involved in plant/bacterium interactions and, notably, able to induce plant systemic resistance. The bud genes, involved in butanediol production, are conserved in the phytopathogenic enterobacteria of the genera Dickeya, Pectobacterium, Erwinia, Pantoea and Brenneria. Inactivation of the bud genes of Dickeya dadantii revealed that the virulence of budA, budB and budR mutants was clearly reduced. The genes budA, budB and budC are highly expressed during plant infection. These data highlight the importance of butanediol metabolism in limiting acidification of the plant tissue during the development of the soft-rot disease caused by pectinolytic enterobacteria. © 2011 Blackwell Publishing Ltd.

  12. Raised serum IgA to common cell envelope antigens supports enterobacterial inductive contribution to pathogenesis of secondary ankylosing spondylitis.

    Science.gov (United States)

    van Bohemen, C G; Weterings, E; Nabbe, A J; Mulder, C J; Goei The, H S; Zanen, H C

    1987-04-01

    Ankylosing spondylitis (AS) is closely associated with the histocompatibility antigen HLA-B27. Pathogenesis of AS is thought to involve interactions between B27 and certain enterobacterial antigens. However, enterobacterial involvement is uncertain and contested by some. The present paper demonstrates raised serum IgA to a common enterobacterial heat modifiable major outer membrane protein (h-momp; Mr 35,000) in active AS (N = 25; IgA = 1485 +/- 20) compared with controls, who were hospital patients without known arthropathies or gastro-intestinal disease (N = 12; IgA = 548 +/- 59). Serum IgG and IgM did not differ statistically. Raised serum IgA to h-momp might indicate enterobacterial antigenic stimulation from the gastro-intestinal tract and thus support an inductive contribution of enterobacterial antigens to the pathogenesis of secondary AS. It does not necessarily imply direct involvement in the pathogenesis of primary AS. H-momp appears to be a convenient tool for serological studies of AS and at present is likely to be more suitable than other bacterial antigens.

  13. Application of amplified fragment length polymorphism fingerprinting for taxonomy and identification of the soft rot bacteria Erwinia carotovora and Erwinia chrysanthemi.

    Science.gov (United States)

    Avrova, Anna O; Hyman, Lizbeth J; Toth, Rachel L; Toth, Ian K

    2002-04-01

    The soft rot bacteria Erwinia carotovora and Erwinia chrysanthemi are important pathogens of potato and other crops. However, the taxonomy of these pathogens, particularly at subspecies level, is unclear. An investigation using amplified fragment length polymorphism (AFLP) fingerprinting was undertaken to determine the taxonomic relationships within this group based on their genetic relatedness. Following cluster analysis on the similarity matrices derived from the AFLP gels, four clusters (clusters 1 to 4) resulted. Cluster 1 contained Erwinia carotovora subsp. carotovora (subclusters 1a and 1b) and Erwinia carotovora subsp. odorifera (subcluster 1c) strains, while cluster 2 contained Erwinia carotovora subsp. atroseptica (subcluster 2a) and Erwinia carotovora subsp. betavasculorum (subcluster 2b) strains. Clusters 3 and 4 contained Erwinia carotovora subsp. wasabiae and E. chrysanthemi strains, respectively. While E. carotovora subsp. carotovora and E. chrysanthemi showed a high level of molecular diversity (23 to 38% mean similarity), E. carotovora subsp. odorifera, E. carotovora subsp. betavasculorum, E. carotovora subsp. atroseptica, and E. carotovora subsp. wasabiae showed considerably less (56 to 76% mean similarity), which may reflect their limited geographical distributions and/or host ranges. The species- and subspecies-specific banding profiles generated from the AFLPs allowed rapid identification of unknown isolates and the potential for future development of diagnostics. AFLP fingerprinting was also found to be more differentiating than other techniques for typing the soft rot erwinias and was applicable to all strain types, including different serogroups.

  14. Investigation of the response to the enterobacterial common antigen after typhoid vaccination

    Directory of Open Access Journals (Sweden)

    Arlete M. Milhomem

    1987-03-01

    Full Text Available Antibodies against the Salmonella typhi enterobacterial common antigen (ECA and the O and H antigens were investigated in sera from healthy male subjects who had been previously vaccinated with the typhoid vaccine. No serological response to ECA was observed. Sera from subjects not previously vaccinated presented titers of ECA hemagglutinins which quantitatively were related to the presence ofH titers, but not to O agglutinins but with no statistical significance. The results are discussed in relation to the possible protective immunological mechanisms in typhoid fever.

  15. 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.

  16. Erwinia carotovora extracellular proteases : characterization and role in soft rot

    OpenAIRE

    Kyöstiö, Sirkka R. M.

    1990-01-01

    Erwinia carotovora subsp. carotovora (Ecc) strain EC14, a Gram-negative bacterium, causes soft rot on several crops, including potato. Maceration of potato tuber tissue is caused by secreted pectolytic enzymes. Other cell-degrading enzymes may also have roles in pathogenesis, including cellulases, phospholipases, and protease(s). The objectives of this research were to (1) characterize Ecc extracellular protease (Prt) and (2) elucidate its role in potato soft rot. A gene enc...

  17. FIRE BLIGHT SUSCEPTIBILITY OF SOME PEAR VARIETIES (ERWINIA AMYLOVORA, BURILL

    Directory of Open Access Journals (Sweden)

    Zsolt Jakab- Ilyefalv

    2012-01-01

    Full Text Available At Bistriţa Fruit Research and Development Station, in a pear collection, planted with 44 varieties, there has been studied the susceptibility to fire blight (Erwinia amylovora,Burill. During the vegetation period, phytosanitary treatments to control the disease have been applied using copper based products (copper sulfate, Champion, Funguran, Kocide. Erwinia amylovora infections have been influenced by the rainfall registered in April (70.9 mm and May (104.7 mm and the temperature fluctuations in April-May. The field observations on Erwinia amylvora attack demonstrate that the pear varieties have a different susceptibility to this dangerous bacterium. Evaluation of attack level in the pear collection was done using an evaluation scale with 9 scores using a reference resistance scale for : ‘Highly resistant’ , ‘Moderately resistant’, ‘Susceptible’, ‘Very susceptible’ cultivars . The most sensitive pear varieties in the collection were: ‘De Noiembrie’, ‘Abatele Fetel’, ’Daciana’, ’Triumf’, ’Williams Boway’, ’Margareta Marillat’, ’Beauty Tomme’, ‘Williams rosu’, ’Aromata de Bistrita’, ’Jeanne d`Arc’, ’Aramiu de Somes’, ’Belle des Arbres’, ’Zorka’ representing 13.64% of the total pear varieties. In several cultivars there have been observed increased symptoms, a very high susceptibility of infections leading to complete scorching of trees: Jeanne d`Arc’, Williams rosu’, ‘Triumf’, ‘Aromata de Bistrita’, ‘Zorka’. Strong attack symtoms were observed at the pear cultivars ‘Cure’, ‘Euras, ’Ciuda’ ‘Highland’, ‘Precoce Morettini’, ’Monica’, ’Cadillac’, ’Juliana’, ’Somesan’, ’Beurré Hardenpont’ these cultivars being susceptible to Erwinia amylovora , representing 40.91 % of total genotypes . Cultivars ‘Untoasa Geoagiu’, ‘Beurre Hardy’, ‘Trivale’ manifested a certain tolerance to Erwinia amylovora , no attack symptoms being

  18. B cells exposed to enterobacterial components suppress development of experimental colitis

    DEFF Research Database (Denmark)

    Schmidt, Esben Gjerløff Wedebye; Larsen, Hjalte List; Kristensen, Nanna Ny

    2012-01-01

    ). RESULTS: We demonstrate that splenic B cells exposed to ebx produce large amounts of IL-10 in vitro and express CD1d and CD5 previously known to be associated with regulatory B cells. In SCID mice transplanted with colitogenic CD4(+) CD25(-) T cells, co-transfer of ebx-B cells significantly suppressed...... development of colitis. Suppression was dependent on B cell-derived IL-10, as co-transfer of IL-10 knockout ebx-B cells failed to suppress colitis. Ebx-B cell-mediated suppression of colitis was associated with a decrease in interferon gamma (IFN-¿)-producing T(H) 1 cells and increased frequencies of Foxp3......-expressing T cells. CONCLUSIONS: These data demonstrate that splenic B cells exposed to enterobacterial components acquire immunosuppressive functions by which they can suppress development of experimental T cell-mediated colitis in an IL-10-dependent way. (Inflamm Bowel Dis 2011;)....

  19. Application of Amplified Fragment Length Polymorphism Fingerprinting for Taxonomy and Identification of the Soft Rot Bacteria Erwinia carotovora and Erwinia chrysanthemi

    OpenAIRE

    Avrova, Anna O.; Hyman, Lizbeth J.; Toth, Rachel L.; Toth, Ian K.

    2002-01-01

    The soft rot bacteria Erwinia carotovora and Erwinia chrysanthemi are important pathogens of potato and other crops. However, the taxonomy of these pathogens, particularly at subspecies level, is unclear. An investigation using amplified fragment length polymorphism (AFLP) fingerprinting was undertaken to determine the taxonomic relationships within this group based on their genetic relatedness. Following cluster analysis on the similarity matrices derived from the AFLP gels, four clusters (c...

  20. 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

  1. First Report of a Human Isolate of Erwinia persicinus

    Science.gov (United States)

    O’Hara, Caroline M.; Steigerwalt, Arnold G.; Hill, Bertha C.; Miller, J. Michael; Brenner, Don J.

    1998-01-01

    Erwinia persicinus was first described in 1990 after being isolated from a variety of fruits and vegetables, including bananas, cucumbers, and tomatoes. In 1994, it was shown to be the causative agent of necrosis of bean pods. We now report the first human isolate of E. persicinus. The strain was isolated from the urine of an 88-year-old woman who presented with a urinary tract infection. By the hydroxyapatite method, DNA from this strain was shown to be 94.5% related at 60°C and 86% related at 75°C to the type strain of E. persicinus. The biochemical profile of E. persicinus is most similar to those of Erwinia rhapontici, Pantoea agglomerans, and Enterobacter species. It is negative in tests for lysine, arginine, ornithine, dulcitol, and urea. It is motile and positive in tests for d-sorbitol and sucrose. It is susceptible to the expanded-spectrum cephalosporins, aminoglycosides, and fluoroquinolones, but it is resistant to ampicillin, ticarcillin, and cefazolin. PMID:9431957

  2. 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.

  3. 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.

  4. 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.

  5. Functional Expression of Enterobacterial O-Polysaccharide Biosynthesis Enzymes in Bacillus subtilis

    Science.gov (United States)

    Schäffer, Christina; Wugeditsch, Thomas; Messner, Paul; Whitfield, Chris

    2002-01-01

    The expression of heterologous bacterial glycosyltransferases is of interest for potential application in the emerging field of carbohydrate engineering in gram-positive organisms. To assess the feasibility of using enzymes from gram-negative bacteria, the functional expression of the genes wbaP (formerly rfbP), wecA (formerly rfe), and wbbO (formerly rfbF) from enterobacterial lipopolysaccharide O-polysaccharide biosynthesis pathways was examined in Bacillus subtilis. WbaP and WecA are initiation enzymes for O-polysaccharide formation, catalyzing the transfer of galactosyl 1-phosphate from UDP-galactose and N-acetylglucosaminyl 1-phosphate from UDP-N-acetylglucosamine, respectively, to undecaprenylphosphate. The WecA product (undecaprenylpyrophosphoryl GlcNAc) is used as an acceptor to which the bifunctional wbbO gene product sequentially adds a galactopyranose and a galactofuranose residue from the corresponding UDP sugars to form a lipid-linked trisaccharide. Genes were cloned into the shuttle vectors pRB374 and pAW10. In B. subtilis hosts, the genes were effectively transcribed under the vegII promoter control of pRB374, but the plasmids were susceptible to rearrangements and deletion. In contrast, pAW10-based constructs, in which genes were cloned downstream of the tet resistance cassette, were stable but yielded lower levels of enzyme activity. In vitro glycosyltransferase assays were performed in Escherichia coli and B. subtilis, using membrane preparations as sources of enzymes and endogenous undecaprenylphosphate as an acceptor. Incorporation of radioactivity from UDP-α-d-14C-sugar into reaction products verified the functionality of WbaP, WecA, and WbbO in either host. Enzyme activities in B. subtilis varied between 20 and 75% of those measured in E. coli. PMID:12324313

  6. Structure of the ribosomal interacting GTPase YjeQ from the enterobacterial species Salmonella typhimurium

    Energy Technology Data Exchange (ETDEWEB)

    Nichols, C. E. [Division of Structural Biology, The Wellcome Trust Centre for Human Genetics, University of Oxford, Roosevelt Drive, Oxford OX3 7BN (United Kingdom); Johnson, C.; Lamb, H. K. [Institute of Cell and Molecular Biosciences, Catherine Cookson Building, Medical School, Framlington Place, Newcastle University, Newcastle-upon-Tyne NE2 4HH (United Kingdom); Lockyer, M. [Arrow Therapeutics Ltd, Britannia House, Trinity Street, Borough, London SE1 1DA (United Kingdom); Charles, I. G. [The Wolfson Institute for Biomedical Research, The Cruciform Building, University College London, Gower Street, London WC1E 6BT (United Kingdom); Hawkins, A. R. [Institute of Cell and Molecular Biosciences, Catherine Cookson Building, Medical School, Framlington Place, Newcastle University, Newcastle-upon-Tyne NE2 4HH (United Kingdom); Stammers, D. K., E-mail: daves@strubi.ox.ac.uk [Division of Structural Biology, The Wellcome Trust Centre for Human Genetics, University of Oxford, Roosevelt Drive, Oxford OX3 7BN (United Kingdom)

    2007-11-01

    The X-ray crystal structure of the GTPase YjeQ from S. typhimurium is presented and compared with those of orthologues from T. maritima and B. subtilis. The YjeQ class of P-loop GTPases assist in ribosome biogenesis and also bind to the 30S subunit of mature ribosomes. YjeQ ribosomal binding is GTP-dependent and thought to specifically direct protein synthesis, although the nature of the upstream signal causing this event in vivo is as yet unknown. The attenuating effect of YjeQ mutants on bacterial growth in Escherichia coli makes it a potential target for novel antimicrobial agents. In order to further explore the structure and function of YjeQ, the isolation, crystallization and structure determination of YjeQ from the enterobacterial species Salmonella typhimurium (StYjeQ) is reported. Whilst the overall StYjeQ fold is similar to those of the previously reported Thematoga maritima and Bacillus subtilis orthologues, particularly the GTPase domain, there are larger differences in the three OB folds. Although the zinc-finger secondary structure is conserved, significant sequence differences alter the nature of the external surface in each case and may reflect varying signalling pathways. Therefore, it may be easier to develop YjeQ-specific inhibitors that target the N- and C-terminal regions, disrupting the metabolic connectivity rather than the GTPase activity. The availability of coordinates for StYjeQ will provide a significantly improved basis for threading Gram-negative orthologue sequences and in silico compound-screening studies, with the potential for the development of species-selective drugs.

  7. In vitro synthesis of a lipid-linked trisaccharide involved in synthesis of enterobacterial common antigen

    International Nuclear Information System (INIS)

    Barr, K.; Nunes-Edwards, P.; Rick, P.D.

    1989-01-01

    The heteropolysaccharide chains of enterobacterial common antigen (ECA) are made up of linear trisaccharide repeat units with the structure----3)-alpha-D-Fuc4NAc-(1----4)- beta-D-ManNAcA-(1----4)-alpha-D-GlcNAc-(1----, where Fuc4NAc is 4-acetamido-4,6-dideoxy-D-galactose, ManNAcA is N-acetyl-D-mannosaminuronic acid, and GlcNAc is N-acetyl-D-glucosamine. The assembly of these chains involves lipid-linked intermediates, and both GlcNAc-pyrophosphorylundecaprenol (lipid I) and ManNAcA-GlcNAc-pyrophosphorylundecaprenol (lipid II) are intermediates in ECA biosynthesis. In this study we demonstrated that lipid II serves as the acceptor of Fuc4NAc residues in the assembly of the trisaccharide repeat unit of ECA chains. Incubation of Escherichia coli membranes with UDP-GlcNAc, UDP-[14C]ManNAcA, and TDP-[3H]Fuc4NAc resulted in the synthesis of a radioactive glycolipid (lipid III) that contained both [14C]ManNAcA and [3H]Fuc4NAc. The oligosaccharide moiety of lipid III was identified as a trisaccharide by gel-permeation chromatography, and the in vitro synthesis of lipid III was dependent on prior synthesis of lipids I and II. Accordingly, the incorporation of [3H]Fuc4NAc into lipid III from the donor TDP-[3H]Fuc4NAc was dependent on the presence of both UDP-GlcNAc and UDP-ManNAcA in the reaction mixtures. In addition, the in vitro synthesis of lipid III was abolished by tunicamycin. Direct conversion of lipid II to lipid III was demonstrated in two-stage reactions in which membranes were initially incubated with UDP-GlcNAc and UDP-[14C]ManNAcA to allow the synthesis of radioactive lipid II

  8. 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 agglomerans 299R should not be classified into the P. agglomerans species. In addition, another strain (Pantoea sp. At_9b) was identified that may represent a novel Pantoea species. We also evaluated the performance of six commonly used housekeeping genes (atpD, carA, gyrB, infB, recA, and rpoB) in phylogenetic inference. A single gene was not enough to obtain a reliable species tree, and it was necessary to use the multilocus sequence analysis of the six marker genes to recover the Erwinia/Pantoea phylogeny.

  9. Combined effects of arthropod herbivores and phytopathogens on plant performance

    DEFF Research Database (Denmark)

    Hauser, Thure Pavlo; Christensen, Stina; Heimes, Christine

    2013-01-01

    1. Many plants are simultaneously attacked by arthropod herbivores and phytopathogens. These may affect each other directly and indirectly, enhancing or reducing the amount of plant resources they each consume. Ultimately, this may reduce or enhance plant performance relative to what should be ex....... However, as interactive impacts also differed among environments and parasite manipulation methods, this suggests that the ability of plants to compensate such losses may depend on environmental conditions and probably also overall infection load.......1. Many plants are simultaneously attacked by arthropod herbivores and phytopathogens. These may affect each other directly and indirectly, enhancing or reducing the amount of plant resources they each consume. Ultimately, this may reduce or enhance plant performance relative to what should...... be expected from the added impacts of herbivore and pathogen when they attack alone. 2. Previous studies have suggested synergistic and antagonistic impacts on plant performance from certain combinations of arthropods and pathogens, for example, synergistic impacts from necrotrophic pathogens together...

  10. [Chitinolytic activity of Bacillus Cohn.--phytopathogenic fungus antagonist].

    Science.gov (United States)

    Aktuganov, G E; Melent'ev, A I; Kuz'mina, L Iu; Galimzianova, N F; Shirokov, A V

    2003-01-01

    Among the 70 tested Bacillus spp. strains antagonistic to phytopathogenic fungi, 19 were found to possess chitinolytic activity when grown on solid media with 0.5% colloidal chitin. The chitinolytic activity of almost all of these 19 strains grown in liquid cultures ranged from 0.1 to 0.3 U/ml. One of the 19 strains exhibited exochitinase activity. In addition to chitinase, two strains also produced chitosanase and one strain, beta-1,3-glucanase. No correlation was found between the antifungal activity of the bacillar strains studied and their ability to synthesize extracellular chitinase. Among the 19 chitinolytic strains, the correlation between these parameters was also low (r x,y = 0.45), although the enzymatic preparations of most of these strains inhibited the growth of the phytopathogenic fungus Helminthosporium sativum.

  11. Cytotoxin production in phytopathogenic and entomopathogenic Serratia marcescens.

    Science.gov (United States)

    Escobar, M M; Carbonell, G V; Beriam, L O; Siqueira, W J; Yano, T

    2001-01-01

    In this work, culture filtrates of entomopathogenic and phytopathogenic Serratia marcescens strains induced cytotoxic effects on CHO, Vero and HEp-2 cell lines. Morphological changes on sensitive cells were characterized by cell rounding and detachment as soon as 30 min of incubation, culminating in cell death after 24 h. The cytotoxic effect was completely neutralized by specific antiserum indicating that occur antigenic similarity among cytotoxins produced by these strains. The toxicity assays on plants showed that the culture supernatants did not provoke any visible morphological change and did not affect their growth. By contrast, the plants treated with bacterial suspension showed disease symptom, such as shriveling and decay of stores bulbus in onion and lettuce plantlets. In conclusion, this study show that phytopathogenic and entomopathogenic S. marcescens may produce a cytototoxin similar to that produced by clinical isolates and it is toxic to different mammalian cell lines. These results are especially important for studies involving this bacterium as biological control agent.

  12. 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

    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....

  13. Bioprospection of yeasts as biocontrol agents against phytopathogenic molds

    Directory of Open Access Journals (Sweden)

    Márcia Maria Rosa-Magri

    2011-02-01

    Full Text Available Yeasts isolated from sugar cane and maize rhizosphere, leaves and stalks were screened against the phytopathogenic molds Colletotrichum sublineolum and Colletotrichum graminicola, both causal agents of the anthracnose disease in sorghum and maize, respectively. Strains identified as Torulaspora globosa and Candida intermedia were able to inhibit the mold growth, with the first species also exhibiting killer activity. No previous report on the application and potentiality of these yeasts as biocontrol agents were found neither the killer phenotype in Torulaspora globosa.

  14. 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...

  15. Cutaneous infection and bactaeremia caused by Erwinia billingiae: a case report

    Directory of Open Access Journals (Sweden)

    M. Prod'homme

    2017-09-01

    Full Text Available Cellulitis and erysipelas are common skin infections usually caused by Staphylococcus aureus and streptococci. Gram-negative rods are rarely implicated. We report here a case of dermohypodermitis and bactaeremia caused by Erwinia billingiae, a Gram-negative bacteria usually pathogenic and epiphytic to pome fruit tree.

  16. 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

  17. Effectieve kolonisatie van aardappelplanten door Dickeya-soorten (Erwinia chrysanthemi) : themanummer fytobacteriologie

    NARCIS (Netherlands)

    Wolf, van der J.M.; Czajkowski, R.L.; Velvis, H.

    2009-01-01

    De bacterieziekten zwartbenigheid en stengelnatrot, veroorzaakt door Pectobacterium en Dickeya (Erwinia)- soorten, berokkenen grote schade aan de pootaardappelteelt. Bij PRI en HZPC wordt onderzoek verricht naar de verspreiding van deze pathogenen tijdens teelt- en (na)oogst. Het was al bekend dat

  18. Besmetting en erwinia-vrij pootgoed uit diverse bronnen : een literatuuroverzicht

    NARCIS (Netherlands)

    Roozen, N.

    1990-01-01

    In dit verslag wordt het risico besproken van diverse potentiële besmettingsbronnen van erwinia-vrij pootgoed. Het doel hiervan is te achterhalen waar de kennis over het risico van de diverse besmettingsbronnen gebreken vertoont en aangevuld dient te worden middels onderzoek. Er zijn criteria

  19. Comparison of specificity and sensitivity of immunochemical and molecular techniques for reliable detection of Erwinia amylovora

    Czech Academy of Sciences Publication Activity Database

    Kokošková, B.; Mráz, Ivan; Hýblová, Jana

    2007-01-01

    Roč. 52, č. 2 (2007), s. 175-182 ISSN 0015-5632 R&D Projects: GA AV ČR(CZ) 1QS500510558 Institutional research plan: CEZ:AV0Z50510513 Keywords : Erwinia amylovora * detection Subject RIV: EE - Microbiology, Virology Impact factor: 0.989, year: 2007

  20. 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

  1. 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...

  2. Beheersing van Erwinia in bolgewassen : agressief snot en witsnot in hyacint, Zantedeschia, Dahlia en andere bloembolgewassen

    NARCIS (Netherlands)

    Doorn, van J.; Vreeburg, P.J.M.; Leeuwen, van P.J.

    2008-01-01

    De afgelopen acht jaar is in toenemende mate bacterierot (zachtrot, agressief rot) opgetreden in bloembolgewassen zoals hyacint, Muscari, Dahlia en iris. Ook in Freesia en Amaryllis zijn problemen; vooral de pootaardappelsector heeft grote schade door met name Erwinia chrysanthemi (agressief snot).

  3. Regulation of pelD and pelE, Encoding Major Alkaline Pectate Lyases in Erwinia chrysanthemi: Involvement of the Main Transcriptional Factors

    Science.gov (United States)

    Rouanet, Carine; Nomura, Kinya; Tsuyumu, Shinji; Nasser, William

    1999-01-01

    The main virulence factors of the phytopathogenic bacterium Erwinia chrysanthemi are pectinases which attack pectin, the major constituent of the plant cell wall. Of these enzymes, the alkaline isoenzyme named PelD in strain 3937 and PelE in strain EC16 has been described as being particularly important, based on virulence studies of plants. Expression of the pelD and pelE genes is tightly modulated by various regulators, including the KdgR repressor and the cyclic AMP-cyclic AMP receptor protein (CRP) activator complex. The use of a lacZ reporter gene allowed us to quantify the repression of E. chrysanthemi 3937 pelD expression exerted by PecS, another repressor of pectinase synthesis. In vitro DNA-protein interaction experiments, centered on the pelD and pelE wild-type or pelE mutated promoter regions, allowed us to define precisely the sequences involved in the binding of these three regulators and of RNA polymerase (RNAP). These studies revealed an unusual binding of the KdgR repressor and suggested the presence of a UP (upstream) element in the pelD and pelE genes. Investigation of the simultaneous binding of CRP, KdgR, PecS, and the RNAP to the regulatory region of the pelD and pelE genes showed that (i) CRP and RNAP bind cooperatively, (ii) PecS partially inhibits binding of the CRP activator and of the CRP-RNAP complex, and (iii) KdgR stabilizes the binding of PecS and prevents transcriptional initiation by RNAP. Taken together, our data suggest that PecS attenuates pelD and pelE expression rather than acting as a true repressor like KdgR. Overall, control of the pelD and pelE genes of E. chrysanthemi appears to be both complex and novel. PMID:10498706

  4. Functional implications of the beta-helical protein fold: differences in chemical and thermal stabilities of Erwinia chrysanthemi EC16 pectate lyases B, C, and E.

    Science.gov (United States)

    Hurlbert, J C; Preston, J F

    2000-09-15

    Colonization of plant tissue by the phytopathogen Erwinia chrysanthemi EC16 is aided by the activities of the pectate lyase isozymes (PLs), which depolymerize the polygalacturonic acid component (PGA) of plant cell walls. The bacterium secretes four pectate lyases (PLa, PLb, PLc, and PLe), two of which, PLc and PLe, have been shown to fold into a similar domain motif, the beta-helix. To understand the rationale behind the evolution and retention of these isoforms, the susceptibilities of pectate lyases B, C, and E to chemical and thermal denaturation and the resulting enzymatic inactivation were examined. With guanidine hydrochloride used as a denaturant, all three pectate lyases denatured with transition midpoint guanidine hydrochloride concentrations (Cm) of 1.3, 1.1, and 1.8 M for PLb, PLc, and PLe, respectively. Lyase activity decreased in direct response to loss of secondary structure in all enzymes. Pectate lyases B and C demonstrated increased enzymatic activity at temperatures above 30 degrees C, with maximal activity observed at 40 degrees C for PLb and 35 degrees C for PLc. Transition midpoints (Tm) as measured by circular dichroism were at 46.9 degrees C for PLb and 44.3 degrees C for PLc, indicating detectable conformational changes accompanying thermal inactivation. Decreased enzymatic activity of PLe was observed at all temperatures above 30 degrees C, and the enzyme was found to possess a Tm at 38.9 degrees C. The data demonstrate structural differences among these enzymes that may be the basis for different enzymatic efficiencies under the potential array of environmental conditions experienced by the bacterium. These differences, in turn, may play a part in the retention of these isozymes as virulence factors, allowing the successful colonization of susceptible plant hosts.

  5. Role of the PhoP-PhoQ system in the virulence of Erwinia chrysanthemi strain 3937: involvement in sensitivity to plant antimicrobial peptides, survival at acid Hh, and regulation of pectolytic enzymes.

    Science.gov (United States)

    Llama-Palacios, Arancha; López-Solanilla, Emilia; Rodríguez-Palenzuela, Pablo

    2005-03-01

    Erwinia chrysanthemi is a phytopathogenic bacterium that causes soft-rot diseases in a broad number of crops. The PhoP-PhoQ system is a key factor in pathogenicity of several bacteria and is involved in the bacterial resistance to different factors, including acid stress. Since E. chrysanthemi is confronted by acid pH during pathogenesis, we have studied the role of this system in the virulence of this bacterium. In this work, we have isolated and characterized the phoP and phoQ mutants of E. chrysanthemi strain 3937. It was found that: (i) they were not altered in their growth at acid pH; (ii) the phoQ mutant showed diminished ability to survive at acid pH; (iii) susceptibility to the antimicrobial peptide thionin was increased; (iv) the virulence of the phoQ mutant was diminished at low and high magnesium concentrations, whereas the virulence of the phoP was diminished only at low magnesium concentrations; (v) in planta Pel activity of both mutant strains was drastically reduced; and (vi) both mutants lagged behind the wild type in their capacity to change the apoplastic pH. These results suggest that the PhoP-PhoQ system plays a role in the virulence of this bacterium in plant tissues, although it does not contribute to bacterial growth at acid pH.

  6. Extracellular Membrane Vesicles and Phytopathogenicity of Acholeplasma laidlawii PG8

    Directory of Open Access Journals (Sweden)

    Vladislav M. Chernov

    2012-01-01

    Full Text Available For the first time, the phytopathogenicity of extracellular vesicles of Acholeplasma laidlawii PG8 (a ubiquitous mycoplasma that is one of the five common species of cell culture contaminants and is a causative agent for phytomycoplasmoses in Oryza sativa L. plants was studied. Data on the ability of extracellular vesicles of Acholeplasma laidlawii PG8 to penetrate from the nutrient medium into overground parts of Oryza sativa L. through the root system and to cause alterations in ultrastructural organization of the plants were presented. As a result of the analysis of ultrathin leaf sections of plants grown in medium with A. laidlawii PG8 vesicles, we detected significant changes in tissue ultrastructure characteristic to oxidative stress in plants as well as their cultivation along with bacterial cells. The presence of nucleotide sequences of some mycoplasma genes within extracellular vesicles of Acholeplasma laidlawii PG8 allowed a possibility to use PCR (with the following sequencing to perform differential detection of cells and bacterial vesicles in samples under study. The obtained data may suggest the ability of extracellular vesicles of the mycoplasma to display in plants the features of infection from the viewpoint of virulence criteria—invasivity, infectivity—and toxigenicity—and to favor to bacterial phytopathogenicity.

  7. The Cultivation of Antagonistic Bacteria in Irradiated Sludge for Biological Control of Soft Rot Erwinias : Screening of Antagonistic Bacteria for biological Control of Soft Rot Erwinias

    International Nuclear Information System (INIS)

    Sermkiattipong, Ng.; Sangsuk, L; Rattanapiriyakul, P; Dejsirilert, S.; Thaveechai, N.

    1998-01-01

    Pure cultures of 57 bacterial isolates for antagonistic activity screening were isolated from three areas of soft rot infested vegetable soil and 58 isolates were obtained from commercial seed compost and seed compost product of Division of Soil and Water Conservation, Department of Land Development. A total of 115 bacterial isolates were evaluated for antagonizing activity against Erwinia carotovora subsp. atroceptica in vitro. Out of them, 18 isolates were antagonists by showing zone of inhibition ranging from 1 to 17 mm by diameter. Most of antagonistic bacteria were identified as Bacillus spp. whereas only one isolate was Pseudomonas vesicularis

  8. Genetic Diversity in Salmonella Isolates from Ducks and their Environments in Penang, Malaysia using Enterobacterial Repetitive Intergenic Consensus

    Directory of Open Access Journals (Sweden)

    Frederick Adzitey 1, Gulam Rusul Rahmat Ali2*, Nurul Huda2 and Rosma Ahmad3

    2013-07-01

    Full Text Available A total of 107 Salmonella isolates (37 S. typhimurium, 26 S. hadar, 15 S. enteritidis, 15 S. braenderup, and 14 S. albany isolated from ducks and their environments in Penang, Malaysia were typed using enterobacterial repetitive intergenic consensus (ERIC to determine their genetic diversity. Analysis of the Salmonella strains by ERIC produced DNA fingerprints of different sizes for differentiation purposes. The DNA fingerprints or band sizes ranged from 14-8300bp for S. Typhimurium, 146-6593bp for S. hadar, 15-4929bp for S. enteritidis, 14-5142bp for S. braenderup and 7-5712bp for S. albany. Cluster analysis at a coefficient of 0.85 grouped the Salmonella strains into various clusters and singletons. S. typhimurium were grouped into 10 clusters and 6 singletons, S. Hadar were grouped into 3 clusters and 18 singletons, S. enteritidis were grouped into 3 clusters and 7 singletons, S. braenderup were grouped into 4 clusters and 7 singletons, and S. albany were grouped into 3 clusters and 7 singletons with discriminatory index (D ranging from 0.92-0.98. ERIC proved to be a useful typing tool for determining the genetic diversity of the duck Salmonella strains.

  9. Isolamento de espécies enterobacterianas em Stomoxys calcitrans Isolation of enterobacterial species in Stomoxys calcitrans

    Directory of Open Access Journals (Sweden)

    Bruno Gomes de Castro

    2008-12-01

    Full Text Available Este estudo teve por objetivo relatar o isolamento de espécies enterobacterianas em segmentos de Stomoxys calcitrans coletadas, em propriedades rurais de dois municípios do Estado do Rio de Janeiro, Brasil. Nove das 28 espécies isoladas e identificadas ainda não haviam sido descritas na mosca dos estábulos. Algumas dessas espécies possuem destacado potencial patogênico ao homem e aos animais ou são oportunistas e outras não foram descritas como causadoras de enfermidades de importância médica ou veterinária.This study aimed to report enterobacterial species recovered from body segments of Stomoxys calcitrans collected in dairy farms from counties of Rio de Janeiro State, Brazil. Nine of the isolated and identified species were not yet described in stable flies, according to the worldwide literature. Some of them are known to present pathogenical potential to man and other animals, while others were not described causing diseases of medical or veterinary importance.

  10. Enterobacterial Small Mobile Sequences Carry Open Reading Frames and are Found Intragenically—Evolutionary Implications for Formation of New Peptides

    Directory of Open Access Journals (Sweden)

    Nicholas Delihas

    2007-01-01

    Full Text Available Intergenic repeat units of 127-bp (RU-1 and 168-bp (RU-2, as well as a newly-found class of 103-bp (RU-3, represent small mobile sequences in enterobacterial genomes present in multiple intergenic regions. These repeat sequences display similarities to eukaryotic miniature inverted-repeat transposable elements (MITE. The RU mobile elements have not been reported to encode amino acid sequences. An in silico approach was used to scan genomes for location of repeat units. RU sequences are found to have open reading frames, which are present in annotated gene loci whereby the RU amino acid sequence is maintained. Gene loci that display repeat units include those that encode large proteins which are part of super families that carry conserved domains and those that carry predicted motifs such as signal peptide sequences and transmembrane domains. A putative exported protein in Y. pestis and a phylogenetically conserved putative inner membrane protein in Salmonella species represent some of the more interesting constructs. We hypothesize that a major outcome of RU open reading frame fusions is the evolutionary emergence of new proteins.

  11. Comparison of pulsed-field gel electrophoresis and enterobacterial repetitive intergenic consensus PCR and biochemical tests to characterize Lactococcus garvieae.

    Science.gov (United States)

    Ture, M; Altinok, I; Capkin, E

    2015-01-01

    Biochemical test, pulsed-field gel electrophoresis (PFGE) and enterobacterial repetitive intergenic consensus sequence PCR (ERIC-PCR) were used to compare 42 strains of Lactococcus garvieae isolated from different regions of Turkey, Italy, France and Spain. Twenty biotypes of L. garvieae were formed based on 54 biochemical tests. ERIC-PCR of genomic DNA from different L. garvieae strains resulted in amplification of multiple fragments of DNA in sizes ranging between 200 and 5000 bp with various band intensities. After cutting DNA with ApaI restriction enzyme and running on the PFGE, 11–22 resolvable bands ranging from 2 to 194 kb were observed. Turkish isolates were grouped into two clusters, and only A58 (Italy) strain was connected with Turkish isolates. Similarities between Turkish, Spanish, Italian and French isolates were Turkey, first lactococcosis occurred in Mugla, and then, it has been spread all over the country. Based on ERIC-PCR, Spanish and Italian strains of L. garvieae were related to Mugla strains. Therefore, after comparing PFGE profiles, ERIC-PCR profiles and phenotypic characteristics of 42 strains of L. garvieae, there were no relationships found between these three typing methods. PFGE method was more discriminative than the other methods. © 2014 John Wiley & Sons Ltd.

  12. 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.

  13. 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.

  14. PENGARUH SUMBER MINERAL TERHADAP PENEKANAN Erwinia carotovora OLEH PSEUDOMONAS PENDAR-FLUOR SECARA IN VITRO

    Directory of Open Access Journals (Sweden)

    Hardian Susilo Addy .

    2012-02-01

    Full Text Available Antimicrobial Stimulation of Fluorescent Pseudomonad to Inhibit Soft-rot Pathogen Caused by Erwinia carotovora subsp. Carotovora. This research was conducted to study effect of mineral sources on inhibition Erwinia carotovora by fluorescent pseudomonad. We used several mineral sources to stimulate antimicrobial substances from fluorescent pseudomonad that responsible to inhibit E. carotovora subsp. carotovora in vitro. The results showed that zinc 0,5 mM were the best to increase antagonistics of fluorescent psudomonad againts E. carotovora. Zinc were increased antimicrobial substances twohold compared with control without stimulant agent. Detection of antimicrobial substance using TLC showed that only one antimicrobial was detected with retention factor (Rf of 0,68 – 0,72. However, identification and characterization of that substance is still needed.

  15. Karakterisasi Erwinia Chrysanthemi Penyebab Penyakit Busuk Bakteri Pada Daun Lidah Buaya (Aloe Vera)

    OpenAIRE

    SUPRIADI,; IBRAHIM, NILDAR; TARYONO,

    2002-01-01

    Characteristics o/Envinia chysanthemi causing bacterial soft rot ofAloe (Aloe VeraJThe bacterial sot rot of aloe, caused by Erwinia chrysanthemi, was first identified in Caibbean Island in 1992. In early 2001, similar symptoms were found on the aloe plants grown in Semplak, Bogor, West Java. Based on its symptom and progressively spread, especially on the leaf and basal stem, it appeared that the disease was serious and therefore threatened the current development of die plants. This study wa...

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

    OpenAIRE

    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 (Pectobacterium carotovorum spp. carotovorum).To identify and detect these soft rot bacterial species in several bulbous ornamentals, standard PCR methods were used. During the last four years, research was dire...

  17. 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. © 2015 The Authors. Microbial Biotechnology published by John Wiley & Sons Ltd and Society for Applied Microbiology.

  18. The phytopathogenic mollicute-insect vector interface: a closer look.

    Science.gov (United States)

    Fletcher, J; Wayadande, A; Melcher, U; Ye, F

    1998-12-01

    ABSTRACT Spiroplasma citri, transmitted by phloem-feeding leafhoppers, moves from the gut lumen through the gut wall, hemolymph, and salivary glands and multiplies in insect tissues. Nontransmissible lines were deficient in their ability to cross these barriers. Molecular analysis revealed extensive chromosomal rearrangements between the transmissible and nontransmissible spiroplasma lines including a large chromosomal inversion and deletions of about 10 kb at each inversion border. One open reading frame of the deleted region, cloned from the transmissible strain BR3-3X, encodes an integral membrane protein of 58 kDa that shares limited sequence similarity with major adhesin proteins of two zoopathogenic mycoplasmas. Adhesion of spiroplasmas to cultured leafhopper cells was inhibited by proteases, suggesting that adherence to host cells is mediated by spiroplasma membrane protein(s). A hypothetical model for insect transmission of phytopathogenic mollicutes is presented.

  19. Evolutionary transitions between beneficial and phytopathogenic Rhodococcus challenge disease management

    Science.gov (United States)

    Thomas, William J; Gordon, Michael I; Stevens, Danielle M; Creason, Allison L; Belcher, Michael S; Serdani, Maryna; Wiseman, Michele S; Grünwald, Niklaus J; Putnam, Melodie L

    2017-01-01

    Understanding how bacteria affect plant health is crucial for developing sustainable crop production systems. We coupled ecological sampling and genome sequencing to characterize the population genetic history of Rhodococcus and the distribution patterns of virulence plasmids in isolates from nurseries. Analysis of chromosome sequences shows that plants host multiple lineages of Rhodococcus, and suggested that these bacteria are transmitted due to independent introductions, reservoir populations, and point source outbreaks. We demonstrate that isolates lacking virulence genes promote beneficial plant growth, and that the acquisition of a virulence plasmid is sufficient to transition beneficial symbionts to phytopathogens. This evolutionary transition, along with the distribution patterns of plasmids, reveals the impact of horizontal gene transfer in rapidly generating new pathogenic lineages and provides an alternative explanation for pathogen transmission patterns. Results also uncovered a misdiagnosed epidemic that implicated beneficial Rhodococcus bacteria as pathogens of pistachio. The misdiagnosis perpetuated the unnecessary removal of trees and exacerbated economic losses. PMID:29231813

  20. Closed Genome Sequence of Phytopathogen Biocontrol Agent Bacillus velezensis Strain AGVL-005, Isolated from Soybean.

    Science.gov (United States)

    Pylro, Victor Satler; Dias, Armando Cavalcante Franco; Andreote, Fernando Dini; Morais, Daniel Kumazawa; Varani, Alessandro de Mello; Andreote, Cristiane Cipolla Fasanella; Bernardo, Eduardo Roberto de Almeida; Zucchi, Tiago

    2018-02-15

    We report here the closed and near-complete genome sequence and annotation of Bacillus velezensis strain AGVL-005, a bacterium isolated from soybean seeds in Brazil and used for phytopathogen biocontrol. Copyright © 2018 Pylro et al.

  1. 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%.

  2. Temperature modulates the secretome of the phytopathogenic fungus Lasiodiplodia theobromae

    Directory of Open Access Journals (Sweden)

    Carina Félix

    2016-08-01

    Full Text Available Environmental alterations modulate host-microorganism interactions. Little is known about how climate changes can trigger pathogenic features on symbiont or mutualistic microorganisms. Current climate models predict increased environmental temperatures.The exposing of phytopathogens to these changing conditions can have particularly relevant consequences for economically important species and for humans. The impact on pathogen/host interaction and the shift on their biogeographical range can induce different levels of virulence in new hosts, allowing massive losses in agricultural and health fields.Lasiodiplodia theobromae is a phytopathogenic fungus responsible for a number of diseases in various plants. It has also been described as an opportunist pathogen in humans, causing infections with different levels of severity. Lasiodiplodia theobromae has a high capacity of adaptation to different environments, such as woody plants, moist argillaceous soils or even humans, being able to grow and infect hosts in a wide range of temperatures (9ºC-39ºC. Nonetheless, the effect of an increase of temperature, as predicted in climate change models, on L. theobromae is unknown.Here we explore the effect of temperature on two strains of L. theobromae – an environmental strain CAA019, and a clinical strain, CBS339.90.We show that both strains are cytotoxic to mammalian cells but while the environmental strain is cytotoxic mainly at 25ºC, the clinical strain is cytotoxic mainly at 30ºC and 37ºC.Extracellular gelatinolytic, xylanolytic, amylolytic and cellulolytic activities at 25ºC and 37ºC were characterized by zymography and the secretome of both strains grown at 25ºC, 30ºC and 37ºC were characterized by electrophoresis and by Orbitrap LC-MS/MS. More than 75% of the proteins were identified, mostly enzymes (glycosyl hydrolases and proteases. The strains showed different protein profiles, which were affected by growth temperature. Also, strain

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

    OpenAIRE

    Ramachandran Kogeethavani; Manaf Uyub Abdul; Zakaria Latiffah

    2015-01-01

    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...

  4. 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...

  5. Characteristics of bacillus strains with antifungal activity against phytopathogens

    Energy Technology Data Exchange (ETDEWEB)

    Lee, Young Keun; Senthilkumar, M. [Korea Atomic Energy Research Institute, Jeongeup (Korea, Republic of)

    2009-12-15

    Four bacterial isolates that showed antifungal activity against Alternaria alternata and other phytopathogens were isolates from bean rhizosphere. 16S rDNA analysis and phylogenetic relationship indicated that these isolates belong to Genus Bacillus. Isolate A1 clustered with Bacillus licheniformis while other isolates A2, A3 and A4 clustered together with B.pumilus. n-Butanol extract of these isolates strongly inhibited the growth of A. alternata while, chloroform extract of isolate A2 and ethyl acetate extract of A1,A3, and A4 inhibited the test fungus partially. All the isolates except A4 produced chitinase enzyme. None of the isolates solubilized mineral phosphate. Radiation sensitivity of isolates A1, A2, A3 and A4 were assessed and the LD{sub 99} values are determined as 0.50, 6.69, 11,60, 1.53 kGy, respectively. Mutant libraries of each isolate were prepared by exposing them to gamma radiation at their respective LD{sub 99} dose. Crude metabolite caused drastic changes on A. alternata hyphal morphology. Appearance of shrunken and collapsed hyphae could be due to the leak of cell wall or changes in membrane permeability.

  6. Characteristics of bacillus strains with antifungal activity against phytopathogens

    International Nuclear Information System (INIS)

    Lee, Young Keun; Senthilkumar, M.

    2009-01-01

    Four bacterial isolates that showed antifungal activity against Alternaria alternata and other phytopathogens were isolates from bean rhizosphere. 16S rDNA analysis and phylogenetic relationship indicated that these isolates belong to Genus Bacillus. Isolate A1 clustered with Bacillus licheniformis while other isolates A2, A3 and A4 clustered together with B.pumilus. n-Butanol extract of these isolates strongly inhibited the growth of A. alternata while, chloroform extract of isolate A2 and ethyl acetate extract of A1,A3, and A4 inhibited the test fungus partially. All the isolates except A4 produced chitinase enzyme. None of the isolates solubilized mineral phosphate. Radiation sensitivity of isolates A1, A2, A3 and A4 were assessed and the LD 99 values are determined as 0.50, 6.69, 11,60, 1.53 kGy, respectively. Mutant libraries of each isolate were prepared by exposing them to gamma radiation at their respective LD 99 dose. Crude metabolite caused drastic changes on A. alternata hyphal morphology. Appearance of shrunken and collapsed hyphae could be due to the leak of cell wall or changes in membrane permeability

  7. Streptomycetes and micromycetes as perspective antagonists of fungal phytopathogens.

    Science.gov (United States)

    Postolaky, O; Syrbu, T; Poiras, N; Baltsat, K; Maslobrod, S; Boortseva, S

    2012-01-01

    Among natural factors that permanently influence on the plants, the soil microorganisms play a special role for the growing of plants as habitants of their rhizosphere. Mainly they are the representatives of actinomycetes genus Streptomyces and fungal genus Penicillium and their metabolic products stimulate plant growth and inhibit the growth of pathogenic fungi and bacteria. The aim of our study was to determine the antagonism of actinomycetes and micromycetes isolated from soils of R. Moldova against the fungal pathogens of agricultural plants. The strains were isolated from 5 types of chernozem (black soil) from central zone of R. Moldova, with different concentration of humus. Most of micromycetes and streptomycetes were isolated from soil sample 1 (monoculture of maize) and soil sample 2 (Poltava road border) with similar humus content (2.4-2.6%). The antifungal activity of micromycetes strains was occurring mostly against Fusarium solani and Thelaviopsis basicola, at streptomycetes against Alternaria alternata and Botrytis cinerea. It was revealed the strains completely inhibit the growth of Alt. alternata (streptomycetes strains 23, 33, 37), B. cinerea (Streptomyces sp. 17), and F. solani (Penicillium sp. 104). Our results allow to consider the actinomycetes Streptomyces sp.9, Streptomyces sp. 12, Streptomyces sp. 17, Streptomyces sp. 37 Streptomyces sp. 66 and micromycetes Penicillium sp. 5, Penicillium sp. 65, Penicillium sp. 104 isolated from soils of R. Moldova, as prospective strains-antagonists against the phytopathogenic fungus, the causative agents of agricultural plants deseasis.

  8. The biosynthesis of brassicicolin A in the phytopathogen Alternaria brassicicola.

    Science.gov (United States)

    Pedras, M Soledade C; Park, Myung Ryeol

    2016-12-01

    Alternaria brassicicola (Schwein.) Wiltshire is a phytopathogenic fungus that together with A. brassicae causes Alternaria black spot disease in Brassica species. Brassicicolin A is the major host-selective phytotoxin produced in cultures of A. brassicicola. Biosynthetic studies to establish the metabolic precursors of brassicicolin A were carried out with isotopically labeled compounds. Incorporation of D-[ 13 C 6 ]glucose, L-[ 15 N]valine, or L-[ 2 H 8 ]valine into brassicicolin A was established using 1 H, 13 C, 15 N NMR and INADEQUATE spectroscopy and HPLC-ESI-MS spectrometry. Based on analyses of the spectroscopic data, the labeling patterns of brassicicolin A isolated from cultures incubated with the labeled precursors are found to be consistent with both the glycolytic and the valine pathways. That is, the carbons of mannitol and acetyl units and the isocyanide carbon atoms are derived from D-[ 13 C 6 ]glucose whereas the hydroxyisopentanoyl and isocyanoisopentanoyl units are derived from L-valine, including the nitrogen atoms of both isocyanide groups. Copyright © 2016 Elsevier Ltd. All rights reserved.

  9. Enterobacteriáceas en partes externas del estadío adulto de Periplaneta americana “cucaracha” capturadas en el mercado Modelo, Iquitos, Perú

    Directory of Open Access Journals (Sweden)

    Xandrix Augusto Ruiz Cabezas

    2015-06-01

    Full Text Available Este estudio, determinó la presencia de Enterobacteriáceas en partes externas del estadío adulto de Periplaneta americana “cucaracha” capturadas en el mercado Modelo de la ciudad de Iquitos. Fueron colectados un total de 68 “cucarachas”, y luego cada una fue depositada en un frasco estéril y trasladadas vivas para el análisis respectivo al laboratorio de Investigación de la Facultad de Ciencias Biológicas; después del aislamiento, las colonias observadas fueron sometidas a pruebas de identificación bioquímica tales como: Prueba de oxidasa, utilización de glucosa, lactosa y sacarosa (agar TSI, pruebas VP-RM, indol, hidrólisis de la úrea. La prevalencia de Enterobacteriáceas se determinó utilizando el método estadístico de distribución de frecuencias y la prueba no paramétrica de Chi cuadrado para prueba de hipótesis. De las 68 cucarachas analizadas se aislaron 118 colonias caracterizadas como lactosa positiva y lactosa negativa, representando el 71 % y 29 % respectivamente de las muestras analizadas.  Finalmente, se aisló e identificó 5 cepas bacterianas, categorizadas en 5 géneros: Providencia (25%, Edwardsiella (9%, Citrobacter (14%, Enterobacter (31% y Klebsiella (21%.

  10. 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.

  11. Thermodependence of growth and enzymatic activities implicated in pathogenicity of two Erwinia carotovora subspecies (Pectobacterium spp.).

    Science.gov (United States)

    Smadja, Bruno; Latour, Xavier; Trigui, Sameh; Burini, Jean François; Chevalier, Sylvie; Orange, Nicole

    2004-01-01

    Erwinia carotovora subsp. atroseptica and Erwinia carotovora subsp. carotovora can cause substantial damage to economically important plant crops and stored products. The occurrence of the disease and the scale of the damage are temperature dependent. Disease development consists first of active multiplication of the bacteria in the infection area and then production of numerous extracellular enzymes. We investigated the effects of various temperatures on these two steps. We assayed the specific growth rate and the pectate lyase and protease activities for eight strains belonging to E. carotovora subsp. atroseptica and E. carotovora subsp. carotovora in vitro. The temperature effect on growth rate and on pectate lyase activity is different for the two subspecies, but protease activity appears to be similarly thermoregulated. Our results are in agreement with ecological data implicating E. carotovora subsp. atroseptica in disease when the temperature is below 20 degrees C. The optimal temperature for pathogenicity appears to be different from the optimal growth temperature but seems to be a compromise between this temperature and temperatures at which lytic activities are maximal.

  12. Two Genomic Regions Involved in Catechol Siderophore Production by Erwinia carotovora

    Science.gov (United States)

    Bull, Carolee T.; Ishimaru, Carol A.; Loper, Joyce E.

    1994-01-01

    Two regions involved in catechol biosynthesis (cbs) of Erwinia carotovora W3C105 were cloned by functional complementation of Escherichia coli mutants that were deficient in the biosynthesis of the catechol siderophore enterobactin (ent). A 4.3-kb region of genomic DNA of E. carotovora complemented the entB402 mutation of E. coli. A second genomic region of 12.8 kb complemented entD, entC147, entE405, and entA403 mutations of E. coli. Although functions encoded by catechol biosynthesis genes (cbsA, cbsB, cbsC, cbsD, and cbsE) of E. carotovora were interchangeable with those encoded by corresponding enterobactin biosynthesis genes (entA, entB, entC, entD, and entE), only cbsE hybridized to its functional counterpart (entE) in E. coli. The cbsEA region of E. carotovora W3C105 hybridized to genomic DNA of 21 diverse strains of E. carotovora but did not hybridize to that of a chrysobactin-producing strain of Erwinia chrysanthemi. Strains of E. carotovora fell into nine groups on the basis of sizes of restriction fragments that hybridized to the cbsEA region, indicating that catechol biosynthesis genes were highly polymorphic among strains of E. carotovora. PMID:16349193

  13. Pantoea agglomerans Strain EH318 Produces Two Antibiotics That Inhibit Erwinia amylovora In Vitro

    Science.gov (United States)

    Wright, Sandra A. I.; Zumoff, Cathy H.; Schneider, Lois; Beer, Steven V.

    2001-01-01

    Pantoea agglomerans (synonym: Erwinia herbicola) strain Eh318 produces through antibiosis a complex zone of inhibited growth in an overlay seeded with Erwinia amylovora, the causal agent of fire blight. This zone is caused by two antibiotics, named pantocin A and B. Using a genomic library of Eh318, two cosmids, pCPP702 and pCPP704, were identified that conferred on Escherichia coli the ability to inhibit growth of E. amylovora. The two cosmids conferred different antibiotic activities on E. coli DH5α and had distinct restriction enzyme profiles. A smaller, antibiotic-conferring DNA segment from each cosmid was cloned. Each subclone was characterized and mutagenized with transposons to generate clones that were deficient in conferring pantocin A and B production, respectively. Mutated subclones were introduced into Eh318 to create three antibiotic-defective marker exchange mutants: strain Eh421 (pantocin A deficient); strain Eh439 (pantocin B deficient), and Eh440 (deficient in both pantocins). Cross-hybridization results, restriction maps, and spectrum-of-activity data using the subclones and marker exchange mutants, supported the presence of two distinct antibiotics, pantocin A and pantocin B, whose biosynthetic genes were present in pCPP702 and pCPP704, respectively. The structure of pantocin A is unknown, whereas that of pantocin B has been determined as (R)-N-[((S)-2-amino-propanoylamino)-methyl]-2-methanesulfonyl-succinamic acid. The two pantocins mainly affect other enteric bacteria, based on limited testing. PMID:11133457

  14. Signaling requirements for Erwinia amylovora-induced disease resistance, callose deposition, and cell growth in the nonhost Arabidopsis thaliana

    Science.gov (United States)

    Erwinia amylovora is the causal agent of the fire blight disease in some plants of the Rosaceae family. The nonhost plant Arabidopsis serves as a powerful system to dissect mechanisms of resistance to E. amylovora. Although not yet known to mount gene-for-gene resistance to E. amylovora, we found ...

  15. Selective inhibition of Erwinia amylovora by the herbicidally active germination-arrest factor (GAF) produced by Pseudomonas bacteria.

    Science.gov (United States)

    Halgren, A; Azevedo, M; Mills, D; Armstrong, D; Thimmaiah, M; McPhail, K; Banowetz, G

    2011-10-01

     The germination-arrest factor (GAF) produced by Pseudomonas fluorescens WH6, and identified as 4-formylaminooxyvinylglycine, specifically inhibits the germination of a wide range of grassy weeds. This study was undertaken to determine whether GAF has antimicrobial activity in addition to its inhibitory effects on grass seed germination. Culture filtrate from Ps. fluorescens WH6 had little or no effect on 17 species of bacteria grown in Petri dish lawns, but the in vitro growth of Erwinia amylovora, the causal agent of the disease of orchard crops known as fire blight, was strongly inhibited by the filtrate. The anti-Erwinia activity of WH6 culture filtrate was shown to be due to its GAF content, and a commercially available oxyvinylglycine, 4-aminoethoxyvinylglycine (AVG), exhibited anti-Erwinia activity similar to that of GAF. The effects of GAF on Erwinia were reversed by particular amino acids. The biological properties of GAF include a rather specific antimicrobial activity against Erw. amylovora. This may be a general property of oxyvinylglycines as AVG exhibited similar activity. The ability of particular amino acids to reverse GAF inhibition is consistent with a potential effect of this compound on the activity of aminotransferases. The results presented here demonstrate a novel antimicrobial activity of oxyvinylglycines and suggest that GAF and/or GAF-producing bacteria may have potential for the control of fire blight. Journal of Applied Microbiology © 2011 The Society for Applied Microbiology. No claim to US Government works.

  16. Symbiotic bacteria (Erwinia sp.) in the gut of Frankliniella occidentalis (Thysanoptera: Thripidae) do not affect its ability to transmit tospovirus

    NARCIS (Netherlands)

    de Vries, E.J.; van de Wetering, F.; van der Hoek, M.M.; Jacobs, G.; Breeuwer, J.A.J.

    2012-01-01

    Tomato spotted wilt virus (TSWV) is one of the most harmful plant viruses and one of its most important vectors is the western flower thrips [Frankliniella occidentalis Pergande (Thysanoptera: Thripidae)]. Recently, we reported the close association of Erwinia sp. gut bacteria with this species of

  17. Characterization of Erwinia amylovora strains from different host plants using repetitive-sequences PCR analysis, and restriction fragment length polymorphism and short-sequence DNA repeats of plasmid pEA29.

    Science.gov (United States)

    Barionovi, D; Giorgi, S; Stoeger, A R; Ruppitsch, W; Scortichini, M

    2006-05-01

    The three main aims of the study were the assessment of the genetic relationship between a deviating Erwinia amylovora strain isolated from Amelanchier sp. (Maloideae) grown in Canada and other strains from Maloideae and Rosoideae, the investigation of the variability of the PstI fragment of the pEA29 plasmid using restriction fragment length polymorphism (RFLP) analysis and the determination of the number of short-sequence DNA repeats (SSR) by DNA sequence analysis in representative strains. Ninety-three strains obtained from 12 plant genera and different geographical locations were examined by repetitive-sequences PCR using Enterobacterial Repetitive Intergenic Consensus, BOX and Repetitive Extragenic Palindromic primer sets. Upon the unweighted pair group method with arithmetic mean analysis, a deviating strain from Amelanchier sp. was analysed using amplified ribosomal DNA restriction analysis (ARDRA) analysis and the sequencing of the 16S rDNA gene. This strain showed 99% similarity to other E. amylovora strains in the 16S gene and the same banding pattern with ARDRA. The RFLP analysis of pEA29 plasmid using MspI and Sau3A restriction enzymes showed a higher variability than that previously observed and no clear-cut grouping of the strains was possible. The number of SSR units reiterated two to 12 times. The strains obtained from pear orchards showing for the first time symptoms of fire blight had a low number of SSR units. The strains from Maloideae exhibit a wider genetic variability than previously thought. The RFLP analysis of a fragment of the pEA29 plasmid would not seem a reliable method for typing E. amylovora strains. A low number of SSR units was observed with first epidemics of fire blight. The current detection techniques are mainly based on the genetic similarities observed within the strains from the cultivated tree-fruit crops. For a more reliable detection of the fire blight pathogen also in wild and ornamentals Rosaceous plants the genetic

  18. Demonstration of strong enterobacterial reactivity of CD4+CD25- T cells from conventional and germ-free mice which is counter-regulated by CD4+CD25+ T cells

    DEFF Research Database (Denmark)

    Gad, Monika; Pedersen, Anders Elm; Kristensen, Nanna N

    2004-01-01

    Unfractionated CD4+ T cells from the gut-associated lymphoid tissue (GALT) and peripheral lymph nodes are unresponsive when exposed to enterobacterial antigens in vitro. Under similar conditions, CD4+ T cells depleted in vivo or in vitro of CD4+CD25+ T cells proliferate extensively. The CD4+CD25- T...

  19. The modality of enterobacterial common antigen polysaccharide chain lengths is regulated by o349 of the wec gene cluster of Escherichia coli K-12.

    Science.gov (United States)

    Barr, K; Klena, J; Rick, P D

    1999-10-01

    The assembly of the phosphoglyceride-linked form of enterobacterial common antigen (ECA(PG)) occurs by a mechanism that involves modulation of polysaccharide chain length. However, the genetic determinant of this modulation has not been identified. Site-directed mutagenesis of o349 of the Escherichia coli K-12 wec gene cluster revealed that this locus encodes a Wzz protein that specifically modulates the chain length of ECA(PG) polysaccharides, and we have designated this locus wzz(ECA). The Wzz(ECA)-mediated modulation of ECA(PG) polysaccharide chains is the first demonstrated example of Wzz regulation involving a polysaccharide that is not linked to the core-lipid A structure of lipopolysaccharide.

  20. The suitability of Finnish climate for fire blight (Erwinia amylovora epidemics on apple

    Directory of Open Access Journals (Sweden)

    Mariela Marinova-Todorova

    2015-03-01

    Full Text Available Fire blight, which is an important disease of apples and pears, has never been detected in continental Finland. In this study the suitability of the Finnish climate for apple blossom blight infections by Erwinia amylovora was evaluated with the epidemiological model MaryblytTM. This was done in fourteen locations, and for two apple cultivars differing in flowering times. Climatic conditions were predicted to be suitable for blossom infections in 18 - 51% of the years, and the annual period of suitable conditions was predicted to last up to two to five days, depending on the location and apple cultivar. The suitable period was predicted to be longer in some locations in central Finland than in those in the southernmost parts of the country. Based on these results the official surveys that are carried out to confirm the absence of fire blight in Finland cannot be targeted only to some parts of the country.

  1. Genetic Characterization Of Syrian Erwinia Amylovora Strains By Amplified Fragment Length Polymorphism Technique

    International Nuclear Information System (INIS)

    Ammouneh, H.; Arabi, M.; Shoaib, A.

    2011-01-01

    Thirty Erwinia amylovora strains, collected from the main rosaceous crop-growing regions in Syria, were chosen as representatives of all major pathogenicity groups and were genetically studied by AFLP. Eight primer combinations were utilized and approximately 300 scorable bands in total were generated. Based on similarity coefficient, E. amylovora strains were placed into a main cluster containing two sub clusters, indicating very low genetic variations among the studied pathogen. The existence of two plasmids, pEA29 (present in nearly all E. amylovora isolates) and pEL60 (present mainly in Lebanese strains), was confirmed using multiplex PCR in all tested Syrian E. amylovora strains, indicating that Lebanese and Syrian isolates may share a common origin.(author)

  2. Expression of a mineral phosphate solubilizing gene from Erwinia herbicola in two rhizobacterial strains.

    Science.gov (United States)

    Rodríguez, H; Gonzalez, T; Selman, G

    2001-11-30

    A genetic construction was carried out using the broad host range vector pKT230 and plasmid pMCG898, which encodes the Erwinia herbicola pyrroloquinoline quinone (PQQ) synthase, a gene involved in mineral phosphate solubilization (mps). The final construction was transformed and expressed in Escherichia coli MC1061, and the recombinant plasmids were transferred to Burkholderia cepacia IS-16 and Pseudomonas sp. PSS recipient cells by conjugation. Clones containing recombinant plasmids produced higher clearing halos in plates with insoluble phosphate as the unique (P) source, in comparison with those of strains without plasmids, demonstrating the heterologous expression of the E. herbicola gene in the recipient strains. This genetic manipulation allowed the increase in mps ability of both strains, enhancing their potentialities as growth promoters of agricultural crops. These results represent the first report on the application of the recombinant DNA methodology for the obtaining of improved phosphate solubilizing ability from rhizobacterial strains for biofertilization purposes.

  3. Detection of bacteriocins produced by plant pathogenic bacteria from the general Erwinia, Pseudomonas and Xanthomonas

    International Nuclear Information System (INIS)

    Biagi, C.M.R. de

    1992-01-01

    Detection of bacteriocin production was studied under distinct conditions using strains of plant pathogenic bacteria from the genera Erwinia, Pseudomonas and Xanthomonas. 58.06%, 79.31% and 40.00% of producing strains were found respectively in the three groups of bacteria using the 523 medium which was the best for the detection of bacteriocin production. Increasing agar concentrations added to the medium up to 1,5% improved the detection. The amount of medium added to the Petri dishes did not affect bacteriocin production. The longest incubation time (72 h.) improved the detection of haloes production. Ultra-violet irradiation in low dosages seems to improve the visualization of haloes production but this is dependent on the tested strains. (author)

  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. 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. Inhibition of Erwinia chrysanthemi growth to different concentrations of folic acid: possible use of folic acid as bacteriostatic agent and fortifying of Solanum tuberosum potato

    Directory of Open Access Journals (Sweden)

    Andrea Marcelo Correa

    2017-05-01

    Full Text Available Introduction:The enterobacteria of the Erwinia spp genus produce disease in potatoes, which is a tuber of mass consumption. The regulation of DNA methylation can regulate the proliferation of Erwinia in such a way that the concentrations of folic acid may have an effect on the microorganism pathogenic ability. On the other hand, the folic acid prevents the appearance of neural tube defects in humans. Objective: To evaluate folic acid as a bacteriostatic agent of Erwinia and, at the same time, as part of the fortification of mass consumption food such as the potatoes. Materials and methods: The biochemical characterization of the Erwinia chrysanthemi was carried out and its growth compared to different concentrations of folic acid was studied. Results: When increasing the concentrations of the vitamin from 0.3 µg/L up to 6.8 µg/L, the bacterial growth of Erwinia chrysanthemi is inhibited. The vitamin inhibits the growth in cultivation of Erwinia chrysanthemi and acts as a bacteriostatic agent. This aspect is of great importance given that, theoretically, if potatoes were fortified with micro-nutrient, this would act against the infectious agent and, at the same time, contribute to the adequate intake of the vitamin in the general population.

  7. Antibacterial activity of some plants from family Apiaceae in relation to selected phytopathogenic bacteria

    OpenAIRE

    Brković, Duško L.; Čomić, Ljiljana; Solujić-Sukdolak, Slavica

    2006-01-01

    Water, ethanol and ethyl acetate extracts of 12 plants from the family Apiaceae were screened for antibacterial activities against selected phytopathogenic bacteria. Following plants were tested: Aegopodium podagraria, Angelica silvestris, Chaerophyllum bulbosum, Daucus carota subsp. carota, Foeniculum vulgare, Heracleum sphondylium, Pastinaca sativa Peucedanum cerevaria, Peucedanum oreoselinum, Pimpinella saxifraga, Sanicula europea, Torilis anthriscus. The antibacterial activities were test...

  8. Role of Proteolytic Enzymes in the Interaction of Phytopathogenic Microorganisms with Plants.

    Science.gov (United States)

    Valueva, T A; Zaichik, B Ts; Kudryavtseva, N N

    2016-12-01

    Various forms of participation of proteolytic enzymes in pathogenesis and defense in plants are reviewed. Along with extracellular proteinases, phytopathogenic microorganisms produce specific effectors having proteolytic activity and capable of acting on proteins inside plant cells. In turn, for defense against pathogens, plants use both extracellular and intracellular proteinases.

  9. Complete genome sequence of phytopathogenic Pectobacterium carotovorum subsp. carotovorum bacteriophage PP1.

    Science.gov (United States)

    Lee, Ju-Hoon; Shin, Hakdong; Ji, Samnyu; Malhotra, Shweta; Kumar, Mukesh; Ryu, Sangryeol; Heu, Sunggi

    2012-08-01

    Pectobacterium carotovorum subsp. carotovorum is a phytopathogen causing soft rot disease on diverse plant species. To control this plant pathogen, P. carotovorum subsp. carotovorum-targeting bacteriophage PP1 was isolated and its genome was completely sequenced to develop a novel biocontrol agent. Interestingly, the 44,400-bp genome sequence does not encode any gene involved in the formation of lysogen, suggesting that this phage may be very useful as a biocontrol agent because it does not make lysogen after host infection. This is the first report on the complete genome sequence of the P. carotovorum subsp. carotovorum-targeting bacteriophage, and it will enhance our understanding of the interaction between phytopathogens and their targeting bacteriophages.

  10. Partitioning of the variance in the growth parameters of Erwinia carotovora on vegetable products.

    Science.gov (United States)

    Shorten, P R; Membré, J-M; Pleasants, A B; Kubaczka, M; Soboleva, T K

    2004-06-01

    The objective of this paper was to estimate and partition the variability in the microbial growth model parameters describing the growth of Erwinia carotovora on pasteurised and non-pasteurised vegetable juice from laboratory experiments performed under different temperature-varying conditions. We partitioned the model parameter variance and covariance components into effects due to temperature profile and replicate using a maximum likelihood technique. Temperature profile and replicate were treated as random effects and the food substrate was treated as a fixed effect. The replicate variance component was small indicating a high level of control in this experiment. Our analysis of the combined E. carotovora growth data sets used the Baranyi primary microbial growth model along with the Ratkowsky secondary growth model. The variability in the microbial growth parameters estimated from these microbial growth experiments is essential for predicting the mean and variance through time of the E. carotovora population size in a product supply chain and is the basis for microbiological risk assessment and food product shelf-life estimation. The variance partitioning made here also assists in the management of optimal product distribution networks by identifying elements of the supply chain contributing most to product variability. Copyright 2003 Elsevier B.V.

  11. Cloning and study of the pectate lyase gene of Erwinia carotovora

    International Nuclear Information System (INIS)

    Bukanov, N.O.; Fonshtein, M.Yu.; Evtushenkov, A.N.; Syarinskii, M.A.; Strel'chenko, P.P.; Yankovski, N.K.; Alikhanyan, S.I.; Fomichev, Yu.K.; Debabov, V.G.

    1986-01-01

    The cloning of the gene of a secretable protein of Erwinia carotovora, pectate lyase, in Escherichia coli was described. Primary cloning was conducted using the phage vector λ 47.1. In the gene library of E. carotovora obtained, eight phages carrying the gene sought were identified according to the appearance of enzymatic activity of the gene product, pectate lyase, in situ. The BamHI fragment of DNA, common to all these phages, was recloned on the plasmid pUC19. It was shown that the cloned pectate lyase gene is represented on the E. carotovora chromosome in one copy. Methods of production of representative gene libraries on phage vectors from no less than 1 μg of cloned DNA even for the genomes of eukaryotes have now been developed. Vectors have been created, for example, λ 47.1, permitting the selection only of hybrid molecules. A number of methods have been developed for the search for a required gene in the library, depending on whether the cloned gene can be expressed or not, and if it can, what properties it will impart to the hybrid clone containing it

  12. 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.

  13. 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

    2015-01-01

    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. PMID:23234799

  14. Antibiosis activity of Pantoea agglomerans biocontrol strain E325 against Erwinia amylovora on apple flower stigmas.

    Science.gov (United States)

    Pusey, P L; Stockwell, V O; Reardon, C L; Smits, T H M; Duffy, B

    2011-10-01

    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 flower stigmas using two antibiosis-deficient mutants. On King's medium B, mutants E325ad1 and E325ad2 have stable smooth-butyrous or hypermucoid colony morphologies, respectively, and the parental strain E325 exhibits phenotypic plasticity with predominantly hypermucoid colonies accompanied by slower-growing, smooth-butyrous colonies. Mutants were tested against E. amylovora on stigmas of detached flowers of crab apple (Malus mandshurica) in growth chambers and apple (Malus domestica) in the orchard. Epiphytic fitness of the antibiosis-negative mutants was similar or greater than the parental strain as determined by relative area under the population curve (RAUPC). In laboratory and orchard trials, both mutants had significantly lower inhibitory activity against the pathogen (i.e., less reduction of E. amylovora RAUPC) compared with the parental strain. E325 and the mutants caused similar decreases in pH in a broth medium, indicating that acidification, which was previously reported as a possible mechanism of pathogen inhibition on stigmas, is not directly related to antibiosis. In this study we provide the first evidence for E325 antibiosis involved in E. amylovora growth suppression on apple flower stigmas.

  15. Cloning, expression, purification and characterisation of Erwinia carotovora L-asparaginase in Escherichia coli

    Directory of Open Access Journals (Sweden)

    Meraj Pourhossein

    2014-01-01

    Full Text Available Background: For the past 30 years, bacterial L-asparaginases have been used as therapeutic agents in the treatment of acute childhood lymphoblastic leukemia. It is found in a variety of organisms such as microbes, plants and mammals. Their intrinsic low-rate glutaminase activity, however, causes serious side-effects, including neurotoxicity, hepatitis, coagulopathy and other dysfunctions. Erwinia carotovora asparaginase shows decreased glutaminase activity, so it is believed to have fewer side-effects in leukemia therapy. Our aim was to clone, express, purify and characterize E. carotovora asparaginase. Materials and Methods: L-asparaginase from E. carotovora NCYC 1526 (ErA was cloned and expressed in Escherichia coli strain BL21 (DE3. The enzyme was purified to homogeneity by affinity chromatography. Various conditions were tested to maximize the production of recombinant asparaginase in E. coli. Results: A new L. asparaginase from E. carotovora NCYC 1526 (ErA was successfully cloned, expressed and purified in E. coli BL21 (DE3. The specific activity of the enzyme was 430 IU/mg. Conclusion: The results of the present work form the basis for a new engineered form of ErA for future therapeutic use, which could be extended with crystallographic studies.

  16. Production of endo-pectate lyase by two stage cultivation of Erwinia carotovora

    Energy Technology Data Exchange (ETDEWEB)

    Fukuoka, Satoshi; Kobayashi, Yoshiaki

    1987-02-26

    The productivity of endo-pectate lyase from Erwinia carotovora GIR 1044 was found to be greatly improved by two stage cultivation: in the first stage the bacterium was grown with an inducing carbon source, e.g., pectin, and in the second stage it was cultivated with glycerol, xylose, or fructose with the addition of monosodium L-glutamate as nitrogen source. In the two stage cultivation using pectin or glycerol as the carbon source the enzyme activity reached 400 units/ml, almost 3 times as much as that of one stage cultivation in a 10 liter fermentor. Using two stage cultivation in the 200 liter fermentor improved enzyme productivity over that in the 10 liter fermentor, with 500 units/ml of activity. Compared with the cultivation in Erlenmeyer flasks, fermentor cultivation improved enzyme productivity. The optimum cultivating conditions were agitation of 480 rpm with aeration of 0.5 vvm at 28 /sup 0/C. (4 figs, 4 tabs, 14 refs)

  17. Complete Genome Sequence of Phytopathogenic Pectobacterium carotovorum subsp. carotovorum Bacteriophage PP1

    OpenAIRE

    Lee, Ju-Hoon; Shin, Hakdong; Ji, Samnyu; Malhotra, Shweta; Kumar, Mukesh; Ryu, Sangryeol; Heu, Sunggi

    2012-01-01

    Pectobacterium carotovorum subsp. carotovorum is a phytopathogen causing soft rot disease on diverse plant species. To control this plant pathogen, P. carotovorum subsp. carotovorum-targeting bacteriophage PP1 was isolated and its genome was completely sequenced to develop a novel biocontrol agent. Interestingly, the 44,400-bp genome sequence does not encode any gene involved in the formation of lysogen, suggesting that this phage may be very useful as a biocontrol agent because it does not m...

  18. Endophytic fungi harbored in the root of Sophora tonkinensis Gapnep: Diversity and biocontrol potential against phytopathogens.

    Science.gov (United States)

    Yao, Yu Qun; Lan, Fang; Qiao, Yun Ming; Wei, Ji Guang; Huang, Rong Shao; Li, Liang Bo

    2017-06-01

    This work, for the first time, investigated the diversity of endophytic fungi harbored in the xylem and phloem of the root of Sophora tonkinensis Gapnep from three geographic localities with emphasis on the influence of the tissue type and geographic locality on endophytic fungal communities and their potential as biocontrol agents against phytopathogens of Panax notoginseng. A total of 655 fungal strains representing 47 taxa were isolated. Forty-two taxa (89.4%) were identified but not five taxa (10.6%) according to morphology and molecular phylogenetics. Out of identifiable taxa, the majority of endophyte taxa were Ascomycota (76.6%), followed by Basidiomycota (8.5%) and Zygomycota (4.3%). The alpha-diversity indices indicated that the species diversity of endophytic fungal community harbored in the root of S. tonkinensis was very high. The colonization and species diversity of endophytic fungal communities were significantly influenced by the geographic locality but not tissue type. The geographic locality and tissue type had great effects on the species composition of endophytic fungal communities. Forty-seven respective strains were challenged by three fungal phytopathogens of P. notoginseng and six strains exhibited significant inhibitory activity. It was noteworthy that endophytic Rhexocercosporidium sp. and F. solani strongly inhibited pathogenic F. solani and other fungal phytopathogens of P. notoginseng. © 2017 The Authors. MicrobiologyOpen published by John Wiley & Sons Ltd.

  19. Biocontrol of Burkholderia cepacia complex bacteria and bacterial phytopathogens by Bdellovibrio bacteriovorus.

    Science.gov (United States)

    McNeely, Damian; Chanyi, Ryan M; Dooley, James S; Moore, John E; Koval, Susan F

    2017-04-01

    Bdellovibrio and like organisms are predatory bacteria that have the unusual property of using the cytoplasmic constituents of other Gram-negative bacteria as nutrients. These predators may thus provide an alternative approach to the biocontrol of human and plant pathogens. Predators were isolated on Burkholderia cenocepacia K56-2 and J2315 as prey cells, in enrichment cultures with soil and sewage. Three isolates (DM7C, DM8A, and DM11A) were identified as Bdellovibrio bacteriovorus on the basis of morphology, a periplasmic life cycle, and 16S rRNA gene sequencing. The prey range of these isolates was tested on Burkholderia cepacia complex bacteria and several phytopathogenic bacteria of agricultural importance. Of 31 strains of the Burkholderia cepacia complex tested, only 4 were resistant to predation by strain DM7C. A subset of 9 of the prey tested were also susceptible to strains DM8A and DM11A. Of 12 phytopathogens tested, 4 were resistant to strains DM7C and DM8A, and only 2 were resistant to strain DM11A. Thus, Bdellovibrio bacteriovorus strains retrieved from environmental samples on 2 Burkholderia cenocepacia isolates from cystic fibrosis patients did not distinguish in their prey range between other isolates of that pathogen or phytopathogens. Such strains hold promise as potential wide-spectrum biocontrol agents.

  20. Antibacterial activity of Artemisia nilagirica leaf extracts against clinical and phytopathogenic bacteria.

    Science.gov (United States)

    Ahameethunisa, Abdul R; Hopper, Waheeta

    2010-01-29

    The six organic solvent extracts of Artemisia nilagirica were screened for the potential antimicrobial activity against phytopathogens and clinically important standard reference bacterial strains. The agar disk diffusion method was used to study the antibacterial activity of A. nilagirica extracts against 15 bacterial strains. The Minimum Inhibitory Concentration (MIC) of the plant extracts were tested using two fold agar dilution method at concentrations ranging from 32 to 512 microg/ml. The phytochemical screening of extracts was carried out for major phytochemical derivatives in A. nilagirica. All the extracts showed inhibitory activity for gram-positive and gram-negative bacteria except for Klebsiella pneumoniae, Enterococcus faecalis and Staphylococcus aureus. The hexane extract was found to be effective against all phytopathogens with low MIC of 32 microg/ml and the methanol extract exhibited a higher inhibition activity against Escherichia coli, Yersinia enterocolitica, Salmonella typhi, Enterobacter aerogenes, Proteus vulgaris, Pseudomonas aeruginosa (32 microg/ml), Bacillus subtilis (64 microg/ml) and Shigella flaxneri (128 microg/ml). The phytochemical screening of extracts answered for the major derivative of alkaloids, amino acids, flavonoids, phenol, quinines, tannins and terpenoids. All the extracts showed antibacterial activity against the tested strains. Of all, methanol and hexane extracts showed high inhibition against clinical and phytopathogens, respectively. The results also indicate the presence of major phytochemical derivatives in the A. nilagirica extracts. Hence, the isolation and purification of therapeutic potential compounds from A. nilagirica could be used as an effective source against bacterial diseases in human and plants.

  1. Enterobacter asburiae sp. nov., a new species found in clinical specimens, and reassignment of Erwinia dissolvens and Erwinia nimipressuralis to the genus Enterobacter as Enterobacter dissolvens comb. nov. and Enterobacter nimipressuralis comb. nov.

    Science.gov (United States)

    Brenner, D J; McWhorter, A C; Kai, A; Steigerwalt, A G; Farmer, J J

    1986-01-01

    Enterobacter asburiae sp. nov. is a new species that was formerly referred to as Enteric Group 17 and that consists of 71 strains, 70 of which were isolated from humans. Enterobacter asburiae sp. nov. strains gave positive reactions in tests for methyl red, citrate utilization (Simmons and Christensen's), urea hydrolysis, L-ornithine decarboxylase, growth in KCN, acid and gas production from D-glucose, and acid production from L-arabinose, cellobiose, glycerol (negative in 1 to 2 days, positive in 3 to 7 days), lactose, D-mannitol, alpha-methyl-D-glucoside, salicin, D-sorbitol, sucrose, trehalose, and D-xylose. They gave negative reactions in the Voges-Proskauer test and in tests for indole, H2S production, phenylalanine, L-lysine decarboxylase, motility, gelatin, utilization of malonate, lipase, DNase, tyrosine clearing, acid production from adonitol, D-arabitol, dulcitol, erythritol, i(myo)-inositol, melibiose, and L-rhamnose. They gave variable reactions in tests for L-arginine dihydrolase (25% positive after 2 days) and acid production from raffinose (69% positive after 2 days). Thirty-four Enterobacter asburiae sp. nov. strains were tested for DNA relatedness by the hydroxyapatite method with 32PO4-labeled DNA from the designated type strain (1497-78, ATCC 35953). The strains were 69 to 100% related in 60 degrees C reactions and 63 to 100% related in 75 degrees C reactions. Divergence within related sequences was 0 to 2.5%. Relatedness of Enterobacter asburiae sp. nov. to 84 strains of members of the Enterobacteriaceae was 5 to 63%, with closest relatedness to strains of Enterobacter cloacae, Erwinia dissolvens, Enterobacter taylorae, Enterobacter agglomerans, Erwinia nimipressuralis, and Enterobacter gergoviae. All strains tested were susceptible to gentamicin and sulfdiazine, and most were susceptible to chloramphenicol, colistin, kanamycin, nalidixic acid, carbenicillin and streptomycin. All strains were resistant to ampicillan, cephalothin, and penicillin

  2. 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

  3. Caracterização de bactérias do género Erwinia causando podridões em plantas ornamentais no Brasil.

    Directory of Open Access Journals (Sweden)

    Irene M.G. Almeida

    1996-05-01

    Full Text Available São descritas as ocorrências de fito-bacterioses em plantios comerciais, ocasionando podridão mole em seis espécies de plantas ornamentais. Testes bioquímicos, fisiológicos, culturais e de patogenicidade permitiram cons-tatar a ocorrência de Erwinia carotovora subsp. carotovora em plantas de comigo-ninguém-pode e orquídea (Phalaenopsis, e de Erwinia chrysanthemi em amarilis, comigo-ninguém-pode, filodendro, orquídeas (Miltonia e Phalaenopsis e violeta africana.

  4. 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 ...

  5. CONTROl.. DE ERWINIA UREDOVORA CON MICROORGANISMOS ANTAGÓNICOS EN CUI..TIVOS DE PYRUS SP~ (PERO)

    OpenAIRE

    De Rico, Myriam E.; Departamento de Microbiología Facultad de Ciencias Pontificia Universidad Javeriana, Bogotá; Parra, Diana P.; Departamento de Microbiología Facultad de Ciencias Pontificia Universidad Javeriana, Bogotá; Baquero, Sandra L.; Departamento de Microbiología Facultad de Ciencias Pontificia Universidad Javeriana, Bogotá

    2013-01-01

    In order to dirninish the use of high doses of the chemical substances employed for the control of fire blight (Erwinia uredovora) in pear crops (Pyrus sp.), antagonistic microorganisms were isolated from the soil near healthy trees, from the rhizosphere, leaves, and stems of the sarne trees. To determine the antagonism in vitro, of these microorganisms against Erwinza uredovora, methods were used: channel, disk and strip. The best results were obtained with the disk method. The microorganism...

  6. Role of motility and chemotaxis in the pathogenesis of Dickeya dadantii 3937 (ex Erwinia chrysanthemi 3937).

    Science.gov (United States)

    Antúnez-Lamas, María; Cabrera-Ordóñez, Ezequiel; López-Solanilla, Emilia; Raposo, Rosa; Trelles-Salazar, Oswaldo; Rodríguez-Moreno, Andrés; Rodríguez-Palenzuela, Pablo

    2009-02-01

    Dickeya dadantii 3937 (ex Erwinia chrysanthemi), a member of the Enterobacteriaceae, causes soft rot in many economically important crops. A successful pathogen has to reach the interior of the plant in order to cause disease. To study the role of motility and chemotaxis in the pathogenicity of D. dadantii 3937, genes involved in the chemotactic signal transduction system (cheW, cheB, cheY and cheZ) and in the structure of the flagellar motor (motA) were mutagenized. All the mutant strains grew like the wild-type in culture media, and the production and secretion of pectolytic enzymes was not affected. As expected, the swimming ability of the mutant strains was reduced with respect to the wild-type: motA (94%), cheY (80%), cheW (74%), cheB (54%) and cheZ (48%). The virulence of the mutant strains was analysed in chicory, Saintpaulia and potato. The mutant strains were also tested for their capability to enter into Arabidopsis leaves. All the mutants showed a significant decrease of virulence in certain hosts; however, the degree of virulence reduction varied depending on the virulence assay. The ability to penetrate Arabidopsis leaves was impaired in all the mutants, whereas the capacity to colonize potato tubers after artificial inoculation was affected in only two mutant strains. In general, the virulence of the mutants could be ranked as motA

  7. 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.

  8. 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.

  9. Floral transmission of Erwinia tracheiphila by cucumber beetles in a wild Cucurbita pepo.

    Science.gov (United States)

    Sasu, M A; Seidl-Adams, I; Wall, K; Winsor, J A; Stephenson, A G

    2010-02-01

    Cucumber beetles, Acalymma vittatum (F.) and Diabrotica undecipunctata howardi (Barber), are specialist herbivores of cucurbits and the vector of Erwinia tracheiphila (E.F. Smith) Holland, the causative agent of wilt disease. Cucumber beetles transmit E. tracheiphila when infected frass falls onto leaf wounds at the site of beetle feeding. We show that E. tracheiphila also can be transmitted via the floral nectaries of Cucurbita pepo ssp. texana L. Andres (Texas gourd). Under field conditions, we found that beetles aggregate in flowers in the late morning, that these beetles chew the anther filaments that cover the nectaries in male flowers thereby exposing the nectary, and that beetle frass accumulates on the nectary. We use real-time polymerase chain reaction to show that most of the flowers produced during the late summer possess beetle frass containing E. tracheiphila. Greenhouse experiments, in which cultures of E. tracheiphila are deposited onto floral nectaries, show that Texas gourds can contract wilt disease through the floral nectaries. Finally, we use green fluorescent protein-transformed E. tracheiphila to document the movement of E. tracheiphila through the nectary into the xylem of the pedicel before the abscission of the flower. Together, these data show that E. tracheiphila can be transmitted through infected frass that falls on or near the floral nectaries. We hypothesize that the concentration of frass from many beetles in the flowers increases both exposure to and the concentration of E. tracheiphila and plays a major role in the dynamics of wilt disease in both wild populations and cultivated squash fields.

  10. Genotypic characterization of Escherichia coli O157:H7 isolates from different sources in the North-West Province, South Africa, using enterobacterial repetitive intergenic consensus PCR analysis.

    Science.gov (United States)

    Ateba, Collins Njie; Mbewe, Moses

    2014-05-30

    In many developing countries, proper hygiene is not strictly implemented when animals are slaughtered and meat products become contaminated. Contaminated meat may contain Escherichia coli (E. coli) O157:H7 that could cause diseases in humans if these food products are consumed undercooked. In the present study, a total of 94 confirmed E. coli O157:H7 isolates were subjected to the enterobacterial repetitive intergenic consensus (ERIC) polymerase chain reaction (PCR) typing to generate genetic fingerprints. The ERIC fragments were resolved by electrophoresis on 2% (w/v) agarose gels. The presence, absence and intensity of band data were obtained, exported to Microsoft Excel (Microsoft Office 2003) and used to generate a data matrix. The unweighted pair group method with arithmetic mean (UPGMA) and complete linkage algorithms were used to analyze the percentage of similarity and matrix data. Relationships between the various profiles and/or lanes were expressed as dendrograms. Data from groups of related lanes were compiled and reported on cluster tables. ERIC fragments ranged from one to 15 per isolate, and their sizes varied from 0.25 to 0.771 kb. A large proportion of the isolates produced an ERIC banding pattern with three duplets ranging in sizes from 0.408 to 0.628 kb. Eight major clusters (I-VIII) were identified. Overall, the remarkable similarities (72% to 91%) between the ERIC profiles for the isolate from animal species and their corresponding food products indicated some form of contamination, which may not exclude those at the level of the abattoirs. These results reveal that ERIC PCR analysis can be reliable in comparing the genetic profiles of E. coli O157:H7 from different sources in the North-West Province of South Africa.

  11. Genotypic Characterization of Escherichia coli O157:H7 Isolates from Different Sources in the North-West Province, South Africa, Using Enterobacterial Repetitive Intergenic Consensus PCR Analysis

    Directory of Open Access Journals (Sweden)

    Collins Njie Ateba

    2014-05-01

    Full Text Available In many developing countries, proper hygiene is not strictly implemented when animals are slaughtered and meat products become contaminated. Contaminated meat may contain Escherichia coli (E. coli O157:H7 that could cause diseases in humans if these food products are consumed undercooked. In the present study, a total of 94 confirmed E. coli O157:H7 isolates were subjected to the enterobacterial repetitive intergenic consensus (ERIC polymerase chain reaction (PCR typing to generate genetic fingerprints. The ERIC fragments were resolved by electrophoresis on 2% (w/v agarose gels. The presence, absence and intensity of band data were obtained, exported to Microsoft Excel (Microsoft Office 2003 and used to generate a data matrix. The unweighted pair group method with arithmetic mean (UPGMA and complete linkage algorithms were used to analyze the percentage of similarity and matrix data. Relationships between the various profiles and/or lanes were expressed as dendrograms. Data from groups of related lanes were compiled and reported on cluster tables. ERIC fragments ranged from one to 15 per isolate, and their sizes varied from 0.25 to 0.771 kb. A large proportion of the isolates produced an ERIC banding pattern with three duplets ranging in sizes from 0.408 to 0.628 kb. Eight major clusters (I–VIII were identified. Overall, the remarkable similarities (72% to 91% between the ERIC profiles for the isolate from animal species and their corresponding food products indicated some form of contamination, which may not exclude those at the level of the abattoirs. These results reveal that ERIC PCR analysis can be reliable in comparing the genetic profiles of E. coli O157:H7 from different sources in the North-West Province of South Africa.

  12. Horizontal Gene Acquisitions, Mobile Element Proliferation, and Genome Decay in the Host-Restricted Plant Pathogen Erwinia Tracheiphila.

    Science.gov (United States)

    Shapiro, Lori R; Scully, Erin D; Straub, Timothy J; Park, Jihye; Stephenson, Andrew G; Beattie, Gwyn A; Gleason, Mark L; Kolter, Roberto; Coelho, Miguel C; De Moraes, Consuelo M; Mescher, Mark C; Zhaxybayeva, Olga

    2016-03-18

    Modern industrial agriculture depends on high-density cultivation of genetically similar crop plants, creating favorable conditions for the emergence of novel pathogens with increased fitness in managed compared with ecologically intact settings. Here, we present the genome sequence of six strains of the cucurbit bacterial wilt pathogen Erwinia tracheiphila (Enterobacteriaceae) isolated from infected squash plants in New York, Pennsylvania, Kentucky, and Michigan. These genomes exhibit a high proportion of recent horizontal gene acquisitions, invasion and remarkable amplification of mobile genetic elements, and pseudogenization of approximately 20% of the coding sequences. These genome attributes indicate that E. tracheiphila recently emerged as a host-restricted pathogen. Furthermore, chromosomal rearrangements associated with phage and transposable element proliferation contribute to substantial differences in gene content and genetic architecture between the six E. tracheiphila strains and other Erwinia species. Together, these data lead us to hypothesize that E. tracheiphila has undergone recent evolution through both genome decay (pseudogenization) and genome expansion (horizontal gene transfer and mobile element amplification). Despite evidence of dramatic genomic changes, the six strains are genetically monomorphic, suggesting a recent population bottleneck and emergence into E. tracheiphila's current ecological niche. © The Author 2016. Published by Oxford University Press on behalf of the Society for Molecular Biology and Evolution.

  13. Horizontal Gene Acquisitions, Mobile Element Proliferation, and Genome Decay in the Host-Restricted Plant Pathogen Erwinia Tracheiphila

    Science.gov (United States)

    Shapiro, Lori R.; Scully, Erin D.; Straub, Timothy J.; Park, Jihye; Stephenson, Andrew G.; Beattie, Gwyn A.; Gleason, Mark L.; Kolter, Roberto; Coelho, Miguel C.; De Moraes, Consuelo M.; Mescher, Mark C.; Zhaxybayeva, Olga

    2016-01-01

    Modern industrial agriculture depends on high-density cultivation of genetically similar crop plants, creating favorable conditions for the emergence of novel pathogens with increased fitness in managed compared with ecologically intact settings. Here, we present the genome sequence of six strains of the cucurbit bacterial wilt pathogen Erwinia tracheiphila (Enterobacteriaceae) isolated from infected squash plants in New York, Pennsylvania, Kentucky, and Michigan. These genomes exhibit a high proportion of recent horizontal gene acquisitions, invasion and remarkable amplification of mobile genetic elements, and pseudogenization of approximately 20% of the coding sequences. These genome attributes indicate that E. tracheiphila recently emerged as a host-restricted pathogen. Furthermore, chromosomal rearrangements associated with phage and transposable element proliferation contribute to substantial differences in gene content and genetic architecture between the six E. tracheiphila strains and other Erwinia species. Together, these data lead us to hypothesize that E. tracheiphila has undergone recent evolution through both genome decay (pseudogenization) and genome expansion (horizontal gene transfer and mobile element amplification). Despite evidence of dramatic genomic changes, the six strains are genetically monomorphic, suggesting a recent population bottleneck and emergence into E. tracheiphila’s current ecological niche. PMID:26992913

  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. Antibacterial activity of Artemisia nilagirica leaf extracts against clinical and phytopathogenic bacteria

    Science.gov (United States)

    2010-01-01

    Background The six organic solvent extracts of Artemisia nilagirica were screened for the potential antimicrobial activity against phytopathogens and clinically important standard reference bacterial strains. Methods The agar disk diffusion method was used to study the antibacterial activity of A. nilagirica extracts against 15 bacterial strains. The Minimum Inhibitory Concentration (MIC) of the plant extracts were tested using two fold agar dilution method at concentrations ranging from 32 to 512 μg/ml. The phytochemical screening of extracts was carried out for major phytochemical derivatives in A. nilagirica. Results All the extracts showed inhibitory activity for gram-positive and gram-negative bacteria except for Klebsiella pneumoniae, Enterococcus faecalis and Staphylococcus aureus. The hexane extract was found to be effective against all phytopathogens with low MIC of 32 μg/ml and the methanol extract exhibited a higher inhibition activity against Escherichia coli, Yersinia enterocolitica, Salmonella typhi, Enterobacter aerogenes, Proteus vulgaris, Pseudomonas aeruginosa (32 μg/ml), Bacillus subtilis (64 μg/ml) and Shigella flaxneri (128 μg/ml). The phytochemical screening of extracts answered for the major derivative of alkaloids, amino acids, flavonoids, phenol, quinines, tannins and terpenoids. Conclusion All the extracts showed antibacterial activity against the tested strains. Of all, methanol and hexane extracts showed high inhibition against clinical and phytopathogens, respectively. The results also indicate the presence of major phytochemical derivatives in the A. nilagirica extracts. Hence, the isolation and purification of therapeutic potential compounds from A. nilagirica could be used as an effective source against bacterial diseases in human and plants. PMID:20109237

  16. Antibacterial activity of Artemisia nilagirica leaf extracts against clinical and phytopathogenic bacteria

    Directory of Open Access Journals (Sweden)

    Hopper Waheeta

    2010-01-01

    Full Text Available Abstract Background The six organic solvent extracts of Artemisia nilagirica were screened for the potential antimicrobial activity against phytopathogens and clinically important standard reference bacterial strains. Methods The agar disk diffusion method was used to study the antibacterial activity of A. nilagirica extracts against 15 bacterial strains. The Minimum Inhibitory Concentration (MIC of the plant extracts were tested using two fold agar dilution method at concentrations ranging from 32 to 512 μg/ml. The phytochemical screening of extracts was carried out for major phytochemical derivatives in A. nilagirica. Results All the extracts showed inhibitory activity for gram-positive and gram-negative bacteria except for Klebsiella pneumoniae, Enterococcus faecalis and Staphylococcus aureus. The hexane extract was found to be effective against all phytopathogens with low MIC of 32 μg/ml and the methanol extract exhibited a higher inhibition activity against Escherichia coli, Yersinia enterocolitica, Salmonella typhi, Enterobacter aerogenes, Proteus vulgaris, Pseudomonas aeruginosa (32 μg/ml, Bacillus subtilis (64 μg/ml and Shigella flaxneri (128 μg/ml. The phytochemical screening of extracts answered for the major derivative of alkaloids, amino acids, flavonoids, phenol, quinines, tannins and terpenoids. Conclusion All the extracts showed antibacterial activity against the tested strains. Of all, methanol and hexane extracts showed high inhibition against clinical and phytopathogens, respectively. The results also indicate the presence of major phytochemical derivatives in the A. nilagirica extracts. Hence, the isolation and purification of therapeutic potential compounds from A. nilagirica could be used as an effective source against bacterial diseases in human and plants.

  17. Complete Genome Sequence of Bacillus subtilis BSn5, an Endophytic Bacterium of Amorphophallus konjac with Antimicrobial Activity for the Plant Pathogen Erwinia carotovora subsp. carotovora ▿

    OpenAIRE

    Deng, Yun; Zhu, Yiguang; Wang, Pengxia; Zhu, Lei; Zheng, Jinshui; Li, Rong; Ruan, Lifang; Peng, Donghai; Sun, Ming

    2011-01-01

    Here, we present the complete genome sequence of Bacillus subtilis strain BSn5, isolated from Amorphophallus konjac calli tissue and showing strong inhibitory activity to Erwinia carotovora subsp. carotovora, which causes Amorphophallus soft rot disease and affects the industry development of this organism.

  18. Complete Genome Sequence of Bacillus subtilis BSn5, an Endophytic Bacterium of Amorphophallus konjac with Antimicrobial Activity for the Plant Pathogen Erwinia carotovora subsp. carotovora ▿

    Science.gov (United States)

    Deng, Yun; Zhu, Yiguang; Wang, Pengxia; Zhu, Lei; Zheng, Jinshui; Li, Rong; Ruan, Lifang; Peng, Donghai; Sun, Ming

    2011-01-01

    Here, we present the complete genome sequence of Bacillus subtilis strain BSn5, isolated from Amorphophallus konjac calli tissue and showing strong inhibitory activity to Erwinia carotovora subsp. carotovora, which causes Amorphophallus soft rot disease and affects the industry development of this organism. PMID:21317323

  19. Complete genome sequence of Bacillus subtilis BSn5, an endophytic bacterium of Amorphophallus konjac with antimicrobial activity for the plant pathogen Erwinia carotovora subsp. carotovora.

    Science.gov (United States)

    Deng, Yun; Zhu, Yiguang; Wang, Pengxia; Zhu, Lei; Zheng, Jinshui; Li, Rong; Ruan, Lifang; Peng, Donghai; Sun, Ming

    2011-04-01

    Here, we present the complete genome sequence of Bacillus subtilis strain BSn5, isolated from Amorphophallus konjac calli tissue and showing strong inhibitory activity to Erwinia carotovora subsp. carotovora, which causes Amorphophallus soft rot disease and affects the industry development of this organism.

  20. [Phytopathogenic bacteria of couch-grass in the crops of wheat].

    Science.gov (United States)

    Iakovleva, L M; Patyka, V F; Gvozdiak, R I; Shcherbina, T N

    2009-01-01

    Bacterialdiseases of weeds in the crops of wheat on the fields of Kyiv and Vinnytsya regions of Ukraine Elytrigia repens (L.) Nevski Agropyrum repens L. were revealed. The following symptoms of bacterial affections: the leaves wither, oval or hatched necrotic spots on green leaves, necroses on the stalks, empty-ears, partial blackening of the ear axes, awns, caryopsises, scales, water-soaked or dark brown with violet shade spots on the rhizomes were found. During the vegetation period bacteria were isolated from the affected plants which caused pathological process in the couch-grass and wheat. The pathogenic bacteria were identified as Pseudomonas syringae, P. viridiflava, Pseudomonas sp., Erwinia carotovora pv. carotovora, Pantoea agglomerans, the part of yellow-pigmentary isolates were not identified. Some Psyringae were isolated from the rhizomes during winterthawing. The paper is presented in Ukrainian.

  1. Characterization of indigoidine biosynthetic genes in Erwinia chrysanthemi and role of this blue pigment in pathogenicity.

    Science.gov (United States)

    Reverchon, Sylvie; Rouanet, Carine; Expert, Dominique; Nasser, William

    2002-02-01

    In the plant-pathogenic bacterium Erwinia chrysanthemi production of pectate lyases, the main virulence determinant, is modulated by a complex network involving several regulatory proteins. One of these regulators, PecS, also controls the synthesis of a blue pigment identified as indigoidine. Since production of this pigment is cryptic in the wild-type strain, E. chrysanthemi ind mutants deficient in indigoidine synthesis were isolated by screening a library of Tn5-B21 insertions in a pecS mutant. These ind mutations were localized close to the regulatory pecS-pecM locus, immediately downstream of pecM. Sequence analysis of this DNA region revealed three open reading frames, indA, indB, and indC, involved in indigoidine biosynthesis. No specific function could be assigned to IndA. In contrast, IndB displays similarity to various phosphatases involved in antibiotic synthesis and IndC reveals significant homology with many nonribosomal peptide synthetases (NRPS). The IndC product contains an adenylation domain showing the signature sequence DAWCFGLI for glutamine recognition and an oxidation domain similar to that found in various thiazole-forming NRPS. These data suggest that glutamine is the precursor of indigoidine. We assume that indigoidine results from the condensation of two glutamine molecules that have been previously cyclized by intramolecular amide bond formation and then dehydrogenated. Expression of ind genes is strongly derepressed in the pecS background, indicating that PecS is the main regulator of this secondary metabolite synthesis. DNA band shift assays support a model whereby the PecS protein represses indA and indC expression by binding to indA and indC promoter regions. The regulatory link, via pecS, between indigoidine and virulence factor production led us to explore a potential role of indigoidine in E. chrysanthemi pathogenicity. Mutants impaired in indigoidine production were unable to cause systemic invasion of potted Saintpaulia ionantha

  2. 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.

  3. Nests of Marsh harrier (Circus aeruginosus L.) as refuges of potentially phytopathogenic and zoopathogenic fungi.

    Science.gov (United States)

    Kornillowicz-Kowalska, Teresa; Kitowski, Ignacy

    2018-01-01

    Birds' nests may be refuges for various species of fungi including that which are potentially phytopathogenic and zoopathogenic. Among the 2449 isolates of fungi obtained from nests of Marsh harriers 96.8% belonged to filamentous fungi. In total, 37 genera were identified from 63 fungi species. Within the mycobiotas of the examined nests populations of fungi which are potentially pathogenic for humans, homoiothermous animals and plants dominated. Among 63 species, 46 (72%) were potentially pathogenic fungi of which 18 species were potentially phytopathogenic and 32 species were pathogenic for homoiothermous animals. Inter alia species of fungi were found in the Marsh harriers nests: Aspergillus fumigatus , Aspergillus flavus , Scopulariopsis brevicaulis , Chrysosporium keratinophilum and Fusarium poae , Fusarium sporotrichioides . In terms of numbers, dominant in Marsh harrier nests were fungi pathogenic to birds, other homoiothermous animals and humans. On that basis it was concluded that Marsh harrier nests are both a source of fungal infections for that species and one of the links in the epidemiological cycle of opportunistic fungi for humans.

  4. Antifungal potential of Bacillus vallismortis R2 against different phytopathogenic fungi

    Directory of Open Access Journals (Sweden)

    Preet K. Kaur

    2015-06-01

    Full Text Available The cash crops grown in an agro-climatic region are prone to infection by various fungal pathogens. The use of chemical fungicides over the years has resulted in emergence of resistant fungal strains, thereby necessitating the development of effective and environmental friendly alternatives. The natural antagonistic interactions among different microbial populations have been exploited as an eco-friendly approach for controlling fungal pathogens resistant to synthetic chemicals. Morphologically distinct bacterial cultures (150, isolated from rhizospheric soils of wheat, rice, onion and tomato plants were screened for their antifungal potential against seven phytopathogenic fungi prevalent in the State of Punjab (India. The bacterial isolate R2, identified as Bacillus vallismortis, supported more than 50% inhibition of different phytopathogenic fungi (Alternaria alternata, Rhizoctonia oryzae, Fusarium oxysporum, Fusarium moniliforme, Colletotrichum sp, Helminthosporium sp and Magnaporthe griseain dual culture plate assay. The thin layer chromatography based bio-autography of acid-precipitated biomolecules (APB indicated the presence of more than one type of antifungal molecule, as evidenced from zones of inhibition against the respective fungal pathogen. The initial analytical studies indicated the presence of surfactin, iturin A and fengycin-like compounds in APB. The antifungal activity of whole cells and APB of isolate R2 was evaluated by light and scanning electron microscopy. The wheat grains treated with APB and exposed to spores of A. alternata showed resistance to the development of black point disease, thereby indicating the potential application of R2 and its biomolecules at field scale level.

  5. Antifungal potential of Bacillus vallismortis R2 against different phytopathogenic fungi

    Energy Technology Data Exchange (ETDEWEB)

    Kaur, P.K.; Kaur, J.; Saini, H.S.

    2015-07-01

    The cash crops grown in an agro-climatic region are prone to infection by various fungal pathogens. The use of chemical fungicides over the years has resulted in emergence of resistant fungal strains, thereby necessitating the development of effective and environmental friendly alternatives. The natural antagonistic interactions among different microbial populations have been exploited as an eco-friendly approach for controlling fungal pathogens resistant to synthetic chemicals. Morphologically distinct bacterial cultures (150), isolated from rhizospheric soils of wheat, rice, onion and tomato plants were screened for their antifungal potential against seven phytopathogenic fungi prevalent in the State of Punjab (India). The bacterial isolate R2, identified as Bacillus vallismortis, supported more than 50% inhibition of different phytopathogenic fungi (Alternaria alternata, Rhizoctonia oryzae, Fusarium oxysporum, Fusarium moniliforme, Colletotrichum sp, Helminthosporium sp and Magnaporthe grisea) in dual culture plate assay. The thin layer chromatography based bio-autography of acid-precipitated biomolecules (APB) indicated the presence of more than one type of antifungal molecule, as evidenced from zones of inhibition against the respective fungal pathogen. The initial analytical studies indicated the presence of surfactin, iturin A and fengycin-like compounds in APB. The antifungal activity of whole cells and APB of isolate R2 was evaluated by light and scanning electron microscopy. The wheat grains treated with APB and exposed to spores of A. alternata showed resistance to the development of black point disease, thereby indicating the potential application of R2 and its biomolecules at field scale level. (Author)

  6. Biological preparation of chitosan nanoparticles and its in vitro antifungal efficacy against some phytopathogenic fungi.

    Science.gov (United States)

    Sathiyabama, M; Parthasarathy, R

    2016-10-20

    The aim of the present study was to prepare Chitosan nanoparticles through biological method with high antifungal activities. Chitosan nanoparticles were prepared by the addition of anionic proteins isolated from Penicillium oxalicum culture to chitosan solutions. The formation of chitosan nanoparticles was preliminary confirmed by UV-vis spectrophotometric analysis. The physico-chemical properties of the chitosan nanoparticles were determined by size and zeta potential analysis, FTIR analysis, HRTEM and XRD pattern. The chitosan nanoparticles were evaluated for its potential to inhibit the growth of phytopathogens viz., Pyricularia grisea, Alternaria solani, Fusarium oxysporum. It is evident from our results that chitosan nanoparticles inhibit the growth of phytopathogens tested. Chitosan nanoparticle treated chickpea seeds showed positive morphological effects such as enhanced germination%, seed vigor index and vegetative biomass of seedlings. All these results indicate that chitosan nanoparticle can be used further under field condition to protect various crops from the devastating fungal pathogens as well as growth promoters. Copyright © 2016 Elsevier Ltd. All rights reserved.

  7. Lipopeptides as main ingredients for inhibition of fungal phytopathogens by Bacillus subtilis/amyloliquefaciens

    Science.gov (United States)

    Cawoy, Hélène; Debois, Delphine; Franzil, Laurent; De Pauw, Edwin; Thonart, Philippe; Ongena, Marc

    2015-01-01

    Some isolates of the Bacillus subtilis/amyloliquefaciens species are known for their plant protective activity against fungal phytopathogens. It is notably due to their genetic potential to form an impressive array of antibiotics including non-ribosomal lipopeptides (LPs). In the work presented here, we wanted to gain further insights into the relative role of these LPs in the global antifungal activity of B. subtilis/amyloliquefaciens. To that end, a comparative study was conducted involving multiple strains that were tested against four different phytopathogens. We combined various approaches to further exemplify that secretion of those LPs is a crucial trait in direct pathogen ward off and this can actually be generalized to all members of these species. Our data illustrate that for each LP family, the fungitoxic activity varies in function of the target species and that the production of iturins and fengycins is modulated by the presence of pathogens. Our data on the relative involvement of these LPs in the biocontrol activity and modulation of their production are discussed in the context of natural conditions in the rhizosphere. PMID:25529983

  8. The Pseudomonas putida T6SS is a plant warden against phytopathogens.

    Science.gov (United States)

    Bernal, Patricia; Allsopp, Luke P; Filloux, Alain; Llamas, María A

    2017-04-01

    Bacterial type VI secretion systems (T6SSs) are molecular weapons designed to deliver toxic effectors into prey cells. These nanomachines have an important role in inter-bacterial competition and provide advantages to T6SS active strains in polymicrobial environments. Here we analyze the genome of the biocontrol agent Pseudomonas putida KT2440 and identify three T6SS gene clusters (K1-, K2- and K3-T6SS). Besides, 10 T6SS effector-immunity pairs were found, including putative nucleases and pore-forming colicins. We show that the K1-T6SS is a potent antibacterial device, which secretes a toxic Rhs-type effector Tke2. Remarkably, P. putida eradicates a broad range of bacteria in a K1-T6SS-dependent manner, including resilient phytopathogens, which demonstrates that the T6SS is instrumental to empower P. putida to fight against competitors. Furthermore, we observed a drastically reduced necrosis on the leaves of Nicotiana benthamiana during co-infection with P. putida and Xanthomonas campestris. Such protection is dependent on the activity of the P. putida T6SS. Many routes have been explored to develop biocontrol agents capable of manipulating the microbial composition of the rhizosphere and phyllosphere. Here we unveil a novel mechanism for plant biocontrol, which needs to be considered for the selection of plant wardens whose mission is to prevent phytopathogen infections.

  9. 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.

  10. Antimicrobial activity of essential oils of Thymus vulgaris and Origanum vulgare on phytopathogenic strains isolated from soybean.

    Science.gov (United States)

    Oliva, M de las M; Carezzano, M E; Giuliano, M; Daghero, J; Zygadlo, J; Bogino, P; Giordano, W; Demo, M

    2015-05-01

    The aim of this work was to study the antimicrobial activity of essential oils obtained from Thymus vulgaris (thyme) and Origanum vulgare (oregano) on phytopathogenic Pseudomonas species isolated from soybean. Strains with characteristics of P. syringae were isolated from leaves of soybean plants with blight symptoms. Ten of these could be identified in Group Ia of LOPAT as P. syringae. Six of these were confirmed as P. syringae using 16S rRNA, indicating the presence of these phytopathogenic bacteria in east and central Argentina. All the phytopathogenic bacteria were re-isolated and identified from the infected plants. MIC values for thyme were 11.5 and 5.7 mg·ml(-1) on P. syringae strains, while oregano showed variability in the inhibitory activity. Both essential oils inhibited all P. syringae strains, with better inhibitory activity than the antibiotic streptomycin. The oils were not bactericidal for all pseudomonads. Both oils contained high carvacrol (29.5% and 19.7%, respectively) and low thymol (1.5%). Natural products obtained from aromatic plants represent potential sources of molecules with biological activity that could be used as new alternatives for the treatment of phytopathogenic bacteria infections. © 2014 German Botanical Society and The Royal Botanical Society of the Netherlands.

  11. Soil metagenomics to identify novel mechanisms of antagonism and antifungal activity for the improved control of phytopathogens

    NARCIS (Netherlands)

    Overbeek, van L.S.; Smalla, K.; Bailey, M.; Jansson, J.; Sjöling, S.; Vogel, T.; Nalin, R.; Elsas, van J.D.

    2008-01-01

    Objective: This project will explore & exploit the anti-phytopathogen diversity present in selected European suppressive soils representing a wide geographical spread. The untapped functional potential present in the unculturable part of the microbiota of these soils will be accessed using

  12. Diffusible and Volatile Antifungal Compounds Produced by an Antagonistic Bacillus velezensis G341 against Various Phytopathogenic Fungi

    Directory of Open Access Journals (Sweden)

    Seong Mi Lim

    2017-10-01

    Full Text Available The aim of this study was to identify volatile and agar-diffusible antifungal metabolites produced by Bacillus sp. G341 with strong antifungal activity against various phytopathogenic fungi. Strain G341 isolated from four-year-old roots of Korean ginseng with rot symptoms was identified as Bacillus velezensis based on 16S rDNA and gyrA sequences. Strain G341 inhibited mycelial growth of all phytopathogenic fungi tested. In vivo experiment results revealed that n-butanol extract of fermentation broth effectively controlled the development of rice sheath blight, tomato gray mold, tomato late blight, wheat leaf rust, barley powdery mildew, and red pepper anthracnose. Two antifungal compounds were isolated from strain G341 and identified as bacillomycin L and fengycin A by MS/MS analysis. Moreover, volatile compounds emitted from strain G341 were found to be able to inhibit mycelial growth of various phytopathogenic fungi. Based on volatile compound profiles of strain G341 obtained through headspace collection and analysis on GC-MS, dimethylsulfoxide, 1-butanol, and 3-hydroxy-2-butanone (acetoin were identified. Taken together, these results suggest that B. valezensis G341 can be used as a biocontrol agent for various plant diseases caused by phytopathogenic fungi.

  13. Diffusible and Volatile Antifungal Compounds Produced by an Antagonistic Bacillus velezensis G341 against Various Phytopathogenic Fungi.

    Science.gov (United States)

    Lim, Seong Mi; Yoon, Mi-Young; Choi, Gyung Ja; Choi, Yong Ho; Jang, Kyoung Soo; Shin, Teak Soo; Park, Hae Woong; Yu, Nan Hee; Kim, Young Ho; Kim, Jin-Cheol

    2017-10-01

    The aim of this study was to identify volatile and agar-diffusible antifungal metabolites produced by Bacillus sp. G341 with strong antifungal activity against various phytopathogenic fungi. Strain G341 isolated from four-year-old roots of Korean ginseng with rot symptoms was identified as Bacillus velezensis based on 16S rDNA and gyrA sequences. Strain G341 inhibited mycelial growth of all phytopathogenic fungi tested. In vivo experiment results revealed that n -butanol extract of fermentation broth effectively controlled the development of rice sheath blight, tomato gray mold, tomato late blight, wheat leaf rust, barley powdery mildew, and red pepper anthracnose. Two antifungal compounds were isolated from strain G341 and identified as bacillomycin L and fengycin A by MS/MS analysis. Moreover, volatile compounds emitted from strain G341 were found to be able to inhibit mycelial growth of various phytopathogenic fungi. Based on volatile compound profiles of strain G341 obtained through headspace collection and analysis on GC-MS, dimethylsulfoxide, 1-butanol, and 3-hydroxy-2-butanone (acetoin) were identified. Taken together, these results suggest that B. valezensis G341 can be used as a biocontrol agent for various plant diseases caused by phytopathogenic fungi.

  14. 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.

  15. Phytopathogenic Dothideomycetes

    NARCIS (Netherlands)

    Crous, P.W.; Verkley, G.J.M.; Groenewald, J.Z.

    2013-01-01

    This volume of Studies in Mycology is dedicated to the plant health officers of the world, who are constantly confronted by a range of plant pathogenic fungi that cause devastating diseases of agricultural and forestry crops. Five main groups of fungi are dealt with, namely Alternaria, Cercospora,

  16. Phytopathogenic Bacteria

    NARCIS (Netherlands)

    Wolf, van der J.M.; Boer, de S.H.

    2015-01-01

    A few hundred bacterial species, belonging to the Proteobacteria, Mollecutes and Actinomycetes cause a large number of different plant diseases, some of which are devastating for agricultural crops. Symptoms of bacterial plant diseases are diverse and include necrosis, tissue maceration, wilting,

  17. Phytopathogenic Nematodes

    NARCIS (Netherlands)

    Helder, J.; Vervoort, M.T.W.; Megen, van H.H.B.; Rybarczyk-Mydlowska, K.; Quist, C.W.; Smant, G.; Bakker, J.

    2015-01-01

    Soil is teeming with life, and rhizosphere soil is even more densely in habited than bulk soil. In terms of biomass, bacteria and fungi are dominant groups, whereas nematodes (roundworms) are the most abundant Metazoans. Bulk soil, soil not directly affected by living plant roots, typically harbours

  18. Antifungal activity of macrofungi extracts on phytopathogenic fungal strains of genera Fusarium sp. and Alternaria sp.

    Directory of Open Access Journals (Sweden)

    Spremo Nemanja R.

    2017-01-01

    Full Text Available During the last decades, intensive application of synthetic fungicides in the agricultural crop protection practice caused growing concern for the existence of toxic chemical residues in food as well as in the whole environment. Instead of using synthetic fungicides, it is suggested that crop protection be carried out by using preparations based on compounds of natural origin (secondary metabolites of plants or microorganisms, including macrofungi from Basidiomycota as biological control agents. The potential of macrofungal species as biocontrol agents was analyzed in this investigation of eight autochthonous species from different locations in Serbia. Both the terricolous species: Coprinus comatus, Coprinellus truncorum, Amanita strobiliformis, Hydnum repandum and the lignicolous species: Flammulina velutipes, Stereum subtomentosum, Trametes versicolor and Bjerkandera adusta were examined, with an aim to detect some novel sources of antifungal agents. This study surveyed antifungal activity of selected macrofungal extracts (MeOH, EtOH and CHCl3 against phytopathogenic Fusarium and Alternaria strains isolated from garlic, soybean and rice: F. proliferatum, F. verticillioides, F. proliferatum, F. graminearum and A. padwickii. Microdilution method in 96 well microplates was applied for the estimation of antifungal effects of macrofungi extracts in the range from 24.75 to 198.00 mg/ml and determination of minimal inhibitory (MIC and minimal fungicidal concentration (MFC. EtOH extract of mychorhizal species H. repandum showed antifungal activity against all analyzed phytopathogenic strains, with the strongest effect on Fusarium strains (MIC 24.75 mg/ml; MFC 24.75 mg/ml. Among others, MeOH extracts of S. subtomentosum and C. micaceus showed similar effects while only B. adusta showed slight effect on Fusarium strains (MIC 24.75-99.00 mg/ml; MFC 24.75-99.00 mg/ml and none effect on A. padwickii. The obtained results indicate the possibility of using

  19. Synthesis and fungistatic activity of new groups of 2,4-dihydroxythiobenzoyl derivatives against phytopathogenic fungi.

    Science.gov (United States)

    Legocki, Jan; Matysiak, Joanna; Niewiadomy, Andrzej; Kostecka, Małgorzata

    2003-01-15

    Twenty-six compounds, derivatives of amides, hydrazines, hydrazides, hydrazones, and semicarbazides, with a 2,4-dihydroxythiobenzoyl moiety, were synthesized from sulfinyl-bis(2,4-dihydroxythiobenzoyl). The compositions and chemical structures of these compounds were confirmed by IR, (1)H NMR, EI-MS, and elemental analysis. Antifungal properties of chemicals under in vitro conditions against five phytopathogenic fungi were estimated. In vivo studies against Erisiphe graminis were also carried out. The compounds N-substituted with an 2,4-dihydroxythiobenzamide group proved to be the most active. N-2-(1-Cinnamylbenzene ester)-2,4-dihydroxythiobenzamide, under in vitro conditions, showed activity at the level of 80-100% development of most pathogens at a concentration of 20 microg/mL and partially at a concentration of 200 microg/mL. For compounds with -HN-NH- or -NH-N= moiety, weak or no fungistatic properties were found at the concentrations studied.

  20. BIOTRANSFORMATION OF FERULIC ACID BY THE PHYTOPATHOGENIC FUNGI Colletotrichum acutatum AND Lasiodiplodia theobromae

    Directory of Open Access Journals (Sweden)

    Manuel Alejandro Numpaque

    2016-01-01

    Full Text Available The microbial transformation of ferulic acid (FA offers a cleaner, more economical alternative for the natural production of flavorings and fragrances. In the present study, the biotransformation of FA using the filamentous phytopathogenic fungi Colletotrichum acutatum and Lasiodiplodia theobromae was researched. Initially, the toxicity of FA against both fungi was evaluated; the FA displayed a moderate toxicity (total inhibition at concentrations ≥ 2000 mg L-1 and apparently a detoxification mechanism was present. Afterwards, the microorganisms were incubated with the substrate at room conditions using a Czapek-Dox culture medium. The results demonstrated that the FA was mainly converted to 4-vinylguaiacol, reaching the highest abundance within the first 48 hours. To a lesser extent, acetovanillone, ethylguaiacol, and vanillin, among others, were produced. Interestingly, the compounds generated in the biotransformation of FA with C. acutatum and L. theobromae have been used as flavorings. Based on the identified metabolites, a possible metabolic pathway was proposed.

  1. The functions of PEX genes in peroxisome biogenesis and pathogenicity in phytopathogenic fungi.

    Science.gov (United States)

    Gao, Fei-Yan; Li, Ling; Wang, Jiao-Yu; Wang, Yan-Li; Sun, Guo-Chang

    2017-10-20

    Peroxisomes are cellular organelles present ubiquitously in eukaryotic cells and are involved in β-oxidation, glyoxylate cycle and a variety of biochemical metabolisms. Recently peroxisomes have been demonstrated to play vital roles in the host infection processes by plant fungal pathogens. The biogenesis of peroxisomes requires a category of proteins named peroxins, which are encoded by the PEX genes. So far, more than 10 PEX genes were isolated in phytopathogenic fungi, and significant research efforts are focused on the mechanism of peroxisome formation and the roles of peroxisome in the development and pathogenicity of fungal pathogens. In this review, we summarize the latest advances in peroxisome biogenesis and functions in pathogenic fungi, including the roles of PEXs in life cycle of peroxisome, peroxisome related metabolisms, and fungal development, infection and pathogenicity, in order to provide references for future studies in plant pathogenic fungi and the control of disease.

  2. A Survey of Phytopathogenic Fungi and Oomycetes in Riyadh, Saudi Arabia

    Directory of Open Access Journals (Sweden)

    Arya Widyawan

    2015-06-01

    Full Text Available A survey of phytopathogenic fungi and Oomyceteswas conducted in Riyadh, Kingdom of Saudi Arabia duringOctober 2008 – May 2009. Total of 223 samples were collectedfrom four regions; Al-Kharj, Oyaynah, Old Diriyah, and Al Amariyah. Isolation was done using Potato Dextrose Agar(PDA. Infected parts were cut then sterilized in chlorox(10%, then were put in petridish that contain PDA andincubated at 25-27 °C. A total twelve genera of fungi andsingle genera of Oomycetes were isolated from the infectedplants and identifi ed as Fusarium spp., Alternaria spp.,Helmintosphorium (Bipolaris spp., Sclerotium spp., Rhizoctoniaspp., Cladosporium spp., Mauginiella scattae, Erysiphe spp.,Leveillula spp., Macrophomina phaseolina, Ustilago spp.,Ulocladium spp., and Phytium spp.

  3. Antifungal Activity of Narceine Methyl Ester and Narceine Isolated from Corydalis longipes Against Some Phytopathogenic Fungi.

    Science.gov (United States)

    Chowdhury, Dibyendu; Maurya, S; Pandey, M B; Pandey, V B; Sarma, B K; Singh, U P

    2005-12-01

    Narceine methyl ester and narceine are potent alkaloids which were isolated from Corydalis longipes were found effective in vitro at very low concentration, i.e., 100~500 ppm against spore germination of some test plant pathogenic fungi (Alternaria solani, A. tagetica, Cercospora abelmoschi, Curvularia maculans, Erysiphe cichoracearum, E. pisi, Fusarium udum, Helminthosporium oryzae, H. penniseti, Ustilago cynodontis). Among the test, phytopathogens the spores of F. udum, C. maculans and H. penniseti were highly sensitive at 200 ppm. However, spores of E. pisi, A. solani and A. tagetica were less sensitive at low concentration followed by other test fungi. Most of the fungi showed zero or nearly zero percent spore germination at 400 and 500 ppm.

  4. Interaction of the psychrotroph Pseudomonas fluorescens In5 with phytopathogens in cold soils

    DEFF Research Database (Denmark)

    Hennessy, Rosanna Catherine; Olsson, Stefan; Stougaard, Peter

    Aim: Potato cultivation in southwest Greenland, at Inneruulalik, omits the use of pesticides while relying on limited crop rotations and despite the presence of plant pathogens in the soil does not suffer from major disease outbreaks. Previously, we have shown that the soil at Inneruulalik...... is disease suppressive owing to the presence of diverse antimicrobial microorganisms. Using culture-based approaches, the psychrotroph Pseudomonas fluorescens In5 was previously isolated from the soil microbiome. The aim of this study is to unravel key biocontrol traits underpinning the antagonistic activity...... of this cold-active bacterium against phytopathogens. Method: A combination of different technologies including genomics, transcriptomics and metabolomics are being used to explore the interaction of the psychrotroph P. fluorescens In5 in dual-culture with diverse plant pathogens. To date, molecular genetics...

  5. Anthelmintic efficacy of gold nanoparticles derived from a phytopathogenic fungus, Nigrospora oryzae.

    Directory of Open Access Journals (Sweden)

    Pradip Kumar Kar

    Full Text Available Exploring a green chemistry approach, this study brings to the fore, the anthelmintic efficacy of gold nanoparticles, highlighting the plausible usage of myconanotechnology. Gold nanoparticles of ∼6 to ∼18 nm diameter were synthesized by treating the mycelia-free culture filtrate of the phytopathogenic fungus with gold chloride. Their size and morphology were confirmed by UV-Vis spectroscopy, DLS data, AFM and TEM images. The XRD studies reveal a crystalline nature of the nanoparticles, which are in cubic phase. The FTIR spectroscopic studies before and after the formation of nanoparticles show the presence of possible functional groups responsible for the bio-reduction and capping of the synthesized gold nanoparticles. The latter were tested as vermifugal agents against a model cestode Raillietina sp., an intestinal parasite of domestic fowl. Further, ultrastructural and biochemical parameters were used to corroborate the efficacy study.

  6. Synthesis and Biological Evaluation of Benzimidazole Phenylhydrazone Derivatives as Antifungal Agents against Phytopathogenic Fungi

    Directory of Open Access Journals (Sweden)

    Xing Wang

    2016-11-01

    Full Text Available A series of benzimidazole phenylhydrazone derivatives (6a–6ai were synthesized and characterized by 1H-NMR, ESI-MS, and elemental analysis. The structure of 6b was further confirmed by single crystal X-ray diffraction as (E-configuration. All the compounds were screened for antifungal activity against Rhizoctonia solani and Magnaporthe oryzae employing a mycelium growth rate method. Compound 6f exhibited significant inhibitory activity against R. solani and M. oryzae with the EC50 values of 1.20 and 1.85 μg/mL, respectively. In vivo testing demonstrated that 6f could effectively control the development of rice sheath blight (RSB and rice blast (RB caused by the above two phytopathogens. This work indicated that the compound 6f with a benzimidazole phenylhydrazone scaffold could be considered as a leading structure for the development of novel fungicides.

  7. Effects of acivicin on growth, mycotoxin production and virulence of phytopathogenic fungi.

    Science.gov (United States)

    Maeda, K; Nakajima, Y; Motoyama, T; Kitou, Y; Kosaki, T; Saito, T; Nishiuchi, T; Kanamaru, K; Osada, H; Kobayashi, T; Kimura, M

    2014-10-01

    Acivicin is an inhibitor of γ-glutamyl transpeptidase and glutamine amidotransferase. When grown on a synthetic minimal agar medium, acivicin strongly inhibited the growth of Magnaporthe oryzae and Alternaria brassicicola, and to a lesser extent, Botrytis cinerea. However, only partial or marginal growth inhibition was observed with regard to Fusarium sporotrichioides and Fusarium graminearum. The growth retardation caused by acivicin was significantly alleviated by cultivating the fungus on a nutrient-rich medium. The inhibition of M. oryzae growth caused by 1 μmol l(-1) of acivicin on minimal agar medium was subdued by the addition of specific single amino acids, including His, a branched-chain amino acid (Leu, Ile or Val), an aromatic amino acid (Trp, Tyr or Phe), Met or Gln, at a concentration of 0·4 mmol l(-1). Trichothecene production by F. graminearum in trichothecene-inducing liquid medium was reduced significantly in the presence of acivicin despite its inability to inhibit growth in the trichothecene-inducing liquid medium. Foliar application of conidia in the presence of acivicin reduced the severity of rice blast disease caused by M. oryzae. These results suggest the usefulness of this modified amino acid natural product to mitigate agricultural problems caused by some phytopathogenic fungi. Significance and impact of the study: Fusarium head blight or scab disease and rice blast, caused by Fusarium graminearum and Magnaporthe oryzae, respectively, are major diseases of cereal crops that cause a significant loss of yield and deterioration in the quality of the grain. The present study investigated the effects of acivicin, a glutamine amino acid analog, on the physiology of various phytopathogenic fungi. Application of acivicin to a fungal culture and conidial suspension reduced mycotoxin production by the wheat scab fungus and the severity of rice blast, respectively. These results suggest the possibility that acivicin may serve as a lead compound to

  8. Laticifer proteins play a defensive role against hemibiotrophic and necrotrophic phytopathogens.

    Science.gov (United States)

    Souza, Diego P; Freitas, Cleverson D T; Pereira, Danielle A; Nogueira, Fábio C; Silva, Fredy D A; Salas, Carlos E; Ramos, Márcio V

    2011-07-01

    Proteins from latex of Calotropis procera (CpLP), Plumeria rubra (PrLP), Carica candamarcensis (P1G10) and Euphorbia tirucalli (EtLP) were tested for antifungal activity against phytopathogens. CpLP and P1G10 inhibited each fungi analyzed. PrLP and EtLP did not exert inhibition. CpLP and P1G10 exhibited preferential inhibitory activity towards R. solani (IC₅₀ = 20.7 and 25.3 µg/ml, respectively). The inhibitory activity was lost after heat treatment or proteolysis, providing evidence for the involvement of proteins in the inhibitory effect. Treatment of CpLP or P1G10 with Dithiothreitol improved both, the endogenous proteolytic activity and the antifungal properties. Conversely, pre-treatment of CpLP or P1G10 with iodoacetamide drastically reduced endogenous proteolytic activities and partially abrogated antifungal activity. Similar results were observed when spores were challenged to germinate in the presence of laticifer proteins. The purified cysteine proteinase CMS2MS2 from Carica candamarcensis latex or papain (E.C. 3.4.22.2), a cysteine proteinase from latex of Carica papaya L., but not trypsin (EC 3.4.21.4) or chymotrypsin (EC 3.4.21.1), two serine proteases, replicated the results obtained with CpLP or P1G10, thus restricting the antifungal property to latex plant cysteine proteinases. CpLP, CMS2MS2 and papain induced production of reactive oxygen species in spores of F. solani, suggesting that inhibition could be linked to oxidative stress. Proteome analysis of CpLP by 2-D electrophoresis and MALDI-TOF-TOF confirmed the existence of various pathogenic-related proteins such as chitinases, peroxidases and osmotins. The results support that laticifer proteins are part of plant defense repertoire against phytopathogenic fungi.

  9. Discovery of Plant Phenolic Compounds That Act as Type III Secretion System Inhibitors or Inducers of the Fire Blight Pathogen, Erwinia amylovora

    OpenAIRE

    Khokhani, Devanshi; Zhang, Chengfang; Li, Yan; Wang, Qi; Zeng, Quan; Yamazaki, Akihiro; Hutchins, William; Zhou, Shan-Shan; Chen, Xin; Yang, Ching-Hong

    2013-01-01

    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...

  10. Identification and Pathogenicity of Phytopathogenic Bacteria Associated with Soft Rot Disease of Girasole Tuber

    Directory of Open Access Journals (Sweden)

    Mamdoh Ewis ISMAIL

    2012-02-01

    Full Text Available During 2010-2011 growing seasons six bacterial isolates were separated from naturally infected girasole plants tubers (Helianthus tuberosus L. cv. Balady, showing soft rot, collected from experimental Farm of the Faculty of Agriculture, in El-Minia University, Egypt. Pathogenicity tests showed various virulence for the bacteria isolated from girasole tubers, found pathogenic. These organisms were characterized as rod-shaped, Gram negative, ?-methyl-d-glucoside medium, reducing substances from sucrose, phos, phatase activity and deep cavities on pectate medium. Otherwise, diagnostic tests suggested that the pathogen was Erwinia carotovora ssp. carotovora. The isolated bacteria caused soft rot of wounded tubers when inoculated into tissues. The bacterial isolates were compared for their degree of pathogenicity as well as for differences in specific symptoms, induced in different hosts. The tested isolates could infect several host ranges, such as fruits of apricot, apple, olive, lemon, squash, eggplant and potato tubers, bulbs and garlic and onion cloves, roots radish, carrot, sweet potato and rape. On the other hand, no symptoms were exhibited on pods of bean and cowpea, faba bean, fruits of pepper and tomato. The extracts of experimentally diseased girasole tubers were active in pectinase and also in caboxymethyl cellulose at pH 6 compared to enzyme activities in healthy tissues. Also, the isolated bacteria increased the total and reducing sugars in infected tissues.

  11. Identification and Pathogenicity of Phytopathogenic Bacteria Associated with Soft Rot Disease of Girasole Tuber

    Directory of Open Access Journals (Sweden)

    Mamdoh Ewis ISMAIL

    2012-02-01

    Full Text Available During 2010-2011 growing seasons six bacterial isolates were separated from naturally infected girasole plants tubers (Helianthus tuberosus L. cv. �Balady�, showing soft rot, collected from experimental Farm of the Faculty of Agriculture, in El-Minia University, Egypt. Pathogenicity tests showed various virulence for the bacteria isolated from girasole tubers, found pathogenic. These organisms were characterized as rod-shaped, Gram negative, ?-methyl-d-glucoside medium, reducing substances from sucrose, phos, phatase activity and deep cavities on pectate medium. Otherwise, diagnostic tests suggested that the pathogen was Erwinia carotovora ssp. carotovora. The isolated bacteria caused soft rot of wounded tubers when inoculated into tissues. The bacterial isolates were compared for their degree of pathogenicity as well as for differences in specific symptoms, induced in different hosts. The tested isolates could infect several host ranges, such as fruits of apricot, apple, olive, lemon, squash, eggplant and potato tubers, bulbs and garlic and onion cloves, roots radish, carrot, sweet potato and rape. On the other hand, no symptoms were exhibited on pods of bean and cowpea, faba bean, fruits of pepper and tomato. The extracts of experimentally diseased girasole tubers were active in pectinase and also in caboxymethyl cellulose at pH 6 compared to enzyme activities in healthy tissues. Also, the isolated bacteria increased the total and reducing sugars in infected tissues.

  12. [Effect of surface-active substances of Acinetobacter calcoaceticus IMV B-7241, Rhodococcus erythropolis IMV Ac-5017, and Nocardia vaccinii K-8 on phytopathogenic bacteria].

    Science.gov (United States)

    Pirog, T P; Konon, A D; Sofilkanich, A P; Iutinskaia, G A

    2013-01-01

    The effect of surface-active substances (SAS's) of Acinetobacter calcoaceticus IMV B-7241, Rhodococcus erythropolis IMV Ac-5017, and Nocardia vaccinii K-8 on phytopathogenic bacteria has been studied. It was shown that the survival of cells (10(5)-10(7) in a milliliter) of the Pseudomonas and Xanthomonas phytopathogenic bacteria was found to be 0-33% after treatment with SAS preparations of the IMV Ac-5017 and IMV B-7241 strains for 2 h (0.15-0.4 mg/mL). In the presence of N. vaccinii K-8 SAS preparations (0.085-0.85 mg/mL), the number of cells of the majority of the studied phytopathogenic bacteria decreased by 95-100%. These data show prospects for using microbial SAS's for the construction of ecologically friendly drugs for regulating the number of phytopathogenic bacteria.

  13. Major phytopathogens and strains from cocoa (Theobroma cacao L.) are differentiated by MALDI-MS lipid and/or peptide/protein profiles.

    Science.gov (United States)

    Dos Santos, Fábio Neves; Tata, Alessandra; Belaz, Kátia Roberta Anacleto; Magalhães, Dilze Maria Argôlo; Luz, Edna Dora Martins Newman; Eberlin, Marcos Nogueira

    2017-03-01

    Phytopathogens are the main disease agents that promote attack of cocoa plantations in all tropical countries. The similarity of the symptoms caused by different phytopathogens makes the reliable identification of the diverse species a challenge. Correct identification is important in the monitoring and management of these pests. Here we show that matrix-assisted laser desorption ionization mass spectrometry (MALDI-MS) in combination with multivariate data analysis is able to rapidly and reliably differentiate cocoa phytopathogens, namely Moniliophthora perniciosa, Phytophthora palmivora, P. capsici, P. citrophthora, P. heveae, Ceratocystis cacaofunesta, C. paradoxa, and C. fimbriata. MALDI-MS reveals unique peptide/protein and lipid profiles which differentiate these phytopathogens at the level of genus, species, and single strain coming from different hosts or cocoa tissues collected in several plantations/places. This fast methodology based on molecular biomarkers is also shown to be sufficiently reproducible and selective and therefore seems to offer a suitable tool to guide the correct application of sanitary defense approaches for infected cocoa plantations. International trading of cocoa plants and products could also be efficiently monitored by MALDI-MS. It could, for instance, prevent the entry of new phytopathogens into a country, e.g., as in the case of Moniliophthora roreri fungus that is present in all cocoa plantations of countries bordering Brazil, but that has not yet attacked Brazilian plantations. Graphical Abstract Secure identification of phytopathogens attacking cocoa plantations has been demonstrated via typical chemical profiles provided by mass spectrometric screening.

  14. [A trial of cultivating Rickettsiella phytoseiuli on the SM IMV-72 medium used for growing phytopathogenic mycoplasmas].

    Science.gov (United States)

    Su'taková, G; Sekejová, Z; Rechacek, J; Skripal', I G; Malinovskaia, L P

    1991-01-01

    The work presents data on growing Rickettsiella phytoseiuli in the SM IMV-72 medium which is used for cultivation of phytopathogenic mycoplasmas. Rickettsiella was observed till the 66th day in the primary cultures and till the 16th day after the first passage. Binary division of the cells has been found only in the primary cultures; a complex cycle of the Rickettsiella development ceased at the stage of formation of the crystal-forming cells. Attempts to passivate Rickettsiella were failure.

  15. Biological relevance of volatile organic compounds emitted during the pathogenic interactions between apple plants and Erwinia amylovora.

    Science.gov (United States)

    Cellini, Antonio; Buriani, Giampaolo; Rocchi, Lorenzo; Rondelli, Elena; Savioli, Stefano; Rodriguez Estrada, Maria T; Cristescu, Simona M; Costa, Guglielmo; Spinelli, Francesco

    2018-01-01

    Volatile organic compounds emitted during the infection of apple (Malus pumila var. domestica) plants by Erwinia amylovora or Pseudomonas syringae pv. syringae were studied by gas chromatography-mass spectrometry and proton transfer reaction-mass spectrometry, and used to treat uninfected plants. Infected plants showed a disease-specific emission of volatile organic compounds, including several bio-active compounds, such as hexenal isomers and 2,3-butanediol. Leaf growth promotion and a higher resistance to the pathogen, expressed as a lower bacterial growth and migration in plant tissues, were detected in plants exposed to volatile compounds from E. amylovora-infected plants. Transcriptional analysis revealed the activation of salicylic acid synthesis and signal transduction in healthy plants exposed to volatiles produced by E. amylovora-infected neighbour plants. In contrast, in the same plants, salicylic acid-dependent responses were repressed after infection, whereas oxylipin metabolism was activated. These results clarify some metabolic and ecological aspects of the pathogenic adaptation of E. amylovora to its host. © 2016 BSPP AND JOHN WILEY & SONS LTD.

  16. 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.

  17. Evidence for the Involvement of an Oxidative Stress in the Initiation of Infection of Pear by Erwinia amylovora1

    Science.gov (United States)

    Venisse, Jean-Stéphane; Gullner, Gabor; Brisset, Marie-Noëlle

    2001-01-01

    Involvement of an oxidative burst, usually related to incompatible plant/pathogen interactions leading to hypersensitive reactions, was investigated with Erwinia amylovora, the causal agent of fire blight of Maloideae subfamily of Rosaceae, in interaction with pear (Pyrus communis; compatible situation) and tobacco (Nicotiana tabacum; incompatible situation). As expected, this necrogenic bacterium induced in tobacco a sustained production of superoxide anion, lipid peroxidation, electrolyte leakage, and concomitant increases of several antioxidative enzymes (ascorbate peroxidases, glutathion reductases, glutathion-S-transferases, and peroxidases), in contrast to the compatible pathogen Pseudomonas syringae pv tabaci, which did not cause such reactions. In pear leaves, however, inoculations with both the disease- and the hypersensitive reaction-inducing bacteria (E. amylovora and P. syringae pv tabaci, respectively) resulted in superoxide accumulation, lipid peroxidation, electrolyte leakage, and enzyme induction at similar rates and according to equivalent time courses. The unexpected ability of E. amylovora to generate an oxidative stress even in compatible situation was linked to its functional hrp (for hypersensitive reaction and pathogenicity) cluster because an Hrp secretion mutant of the bacteria did not induce any plant response. It is suggested that E. amylovora uses the production of reactive oxygen species as a tool to provoke host cell death during pathogenesis to invade plant tissues. The bacterial exopolysaccharide could protect this pathogen against the toxic effects of oxygen species since a non-capsular mutant of E. amylovora induced locally the same responses than the wild type but was unable to further colonize the plant. PMID:11299395

  18. 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.

  19. 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.

  20. Bioactivity of essential oils in phytopathogenic and post-harvest fungi control.

    Science.gov (United States)

    Santamarina, M P; Ibáñez, M D; Marqués, M; Roselló, J; Giménez, S; Blázquez, M A

    2017-11-01

    Commercial thyme and lavender essential oils were analysed by GC/MS. Sixty-six compounds accounting for 98.6-99.6% of total essential oil were identified. Thymol (52.14 ± 0.21%), followed by p-cymene (32.24 ± 0.16%), carvacrol (3.71 ± 0.01%) and γ-terpinene (3.34 ± 0.02%), were the main compounds in thyme essential oil, while large amounts of oxygenated monoterpenes linalool acetate (37.07 ± 0.24%) and linalool (30.16 ± 0.06%) were found in lavender one. In vitro antifungal activity of the essential oils was evaluated at 200 and 300 μg/mL against 10 phytopathogenic and post-harvest fungi, which significantly affect agriculture. Micelial growth inhibition was calculated for each tested fungus and dose. Thyme essential oil showed satisfactory results with 90-100% growth inhibition in almost all the assayed fungi at 300 μg/mL, while lavender essential oil showed no noteworthy inhibition data at either dose, and its growth was even enhanced. Thyme essential oil represents a natural alternative to control harvest and post-harvest fungi, and to extend the shelf-life of agriculture products.

  1. In Vitro Antifungal Activity of Burkholderia gladioli pv. agaricicola against Some Phytopathogenic Fungi

    Directory of Open Access Journals (Sweden)

    Hazem S. Elshafie

    2012-12-01

    Full Text Available The trend to search novel microbial natural biocides has recently been increasing in order to avoid the environmental pollution from use of synthetic pesticides. Among these novel natural biocides are the bioactive secondary metabolites of Burkholderia gladioli pv. agaricicola (Bga. The aim of this study is to determine antifungal activity of Bga strains against some phytopathogenic fungi. The fungicidal tests were carried out using cultures and cell-free culture filtrates against Botrytis cinerea, Aspergillus flavus, Aspergillus niger, Penicillium digitatum, Penicillium expansum, Sclerotinia sclerotiorum and Phytophthora cactorum. Results demonstrated that all tested strains exert antifungal activity against all studied fungi by producing diffusible metabolites which are correlated with their ability to produce extracellular hydrolytic enzymes. All strains significantly reduced the growth of studied fungi and the bacterial cells were more bioactive than bacterial filtrates. All tested Bulkholderia strains produced volatile organic compounds (VOCs, which inhibited the fungal growth and reduced the growth rate of Fusarium oxysporum and Rhizoctonia solani. GC/MS analysis of VOCs emitted by strain Bga 11096 indicated the presence of a compound that was identified as 1-methyl-4-(1-methylethenyl-cyclohexene, a liquid hydrocarbon classified as cyclic terpene. This compound could be responsible for the antifungal activity, which is also in agreement with the work of other authors.

  2. Screening of endophytic fungi with anti-phytopathogen activities from Heptacodium miconioides.

    Science.gov (United States)

    Wang, Lijun; Chen, Tingting; Zou, Yuanyuan; Zhu, Manli; Yu, Wenhua; Zhang, Yinglao

    2015-01-04

    To find anti-phytopathogen compounds from endophytic fungi associated with the endangered species Heptacodium miconioides. Fungi from H. miconioides with antifungal activities were isolated according to the plate growth inhibition method. The fungus with preferable antifungal activities was identified by morphological identification and 5. 8S rRNA sequence analysis. The bioactive metabolites were isolated and purified by chromatographic methods; the structures were determined by spectroscopic analysis. Alternaria solani QZH 10 showed better antifungal activity against Rhizoctorzia solani and Valsa mali with the inhibition rates of 89.1% and 67.9%, respectively. The ethyl acetate crude extract of QZH 10 had strong antifungal activity against Magnaporthe oryzae with the rate of 100. 0% under the concentration of 100 μg/mL. Two antifungal metabolites altersolanol A and 6-O-methylalaternin were isolated and determined from QZH 10. Altersolanol A possessed strong activity against M. oryzae with the inhibition rate of more than 85%, 6-O-methylalaternin had the mightily activity against V. mali with the inhibition rate of 100.0% under the concentration of 100 μg/mL. Altersolanol A and 6-O-methylalaternin are potential fungicides originated from microorganisms.

  3. Volatile compounds emitted by diverse phytopathogenic microorganisms promote plant growth and flowering through cytokinin action.

    Science.gov (United States)

    Sánchez-López, Ángela María; Baslam, Marouane; De Diego, Nuria; Muñoz, Francisco José; Bahaji, Abdellatif; Almagro, Goizeder; Ricarte-Bermejo, Adriana; García-Gómez, Pablo; Li, Jun; Humplík, Jan F; Novák, Ondřej; Spíchal, Lukáš; Doležal, Karel; Baroja-Fernández, Edurne; Pozueta-Romero, Javier

    2016-12-01

    It is known that volatile emissions from some beneficial rhizosphere microorganisms promote plant growth. Here we show that volatile compounds (VCs) emitted by phylogenetically diverse rhizosphere and non-rhizhosphere bacteria and fungi (including plant pathogens and microbes that do not normally interact mutualistically with plants) promote growth and flowering of various plant species, including crops. In Arabidopsis plants exposed to VCs emitted by the phytopathogen Alternaria alternata, changes included enhancement of photosynthesis and accumulation of high levels of cytokinins (CKs) and sugars. Evidence obtained using transgenic Arabidopsis plants with altered CK status show that CKs play essential roles in this phenomenon, because growth and flowering responses to the VCs were reduced in mutants with CK-deficiency (35S:AtCKX1) or low receptor sensitivity (ahk2/3). Further, we demonstrate that the plant responses to fungal VCs are light-dependent. Transcriptomic analyses of Arabidopsis leaves exposed to A. alternata VCs revealed changes in the expression of light- and CK-responsive genes involved in photosynthesis, growth and flowering. Notably, many genes differentially expressed in plants treated with fungal VCs were also differentially expressed in plants exposed to VCs emitted by the plant growth promoting rhizobacterium Bacillus subtilis GB03, suggesting that plants react to microbial VCs through highly conserved regulatory mechanisms. © 2016 John Wiley & Sons Ltd.

  4. Synthesis and characterization of silver nanoparticles: effect on phytopathogen Colletotrichum gloesporioides

    Energy Technology Data Exchange (ETDEWEB)

    Aguilar-Mendez, Miguel A., E-mail: maguilarme@ipn.mx; San Martin-Martinez, Eduardo; Ortega-Arroyo, Lesli [Instituto Politecnico Nacional, Centro de Investigacion en Ciencia Aplicada y Tecnologia Avanzada (Mexico); Cobian-Portillo, Georgina [Instituto Politecnico Nacional, Centro Interdisciplinario de Investigacion para el Desarrollo Integral Regional (Mexico); Sanchez-Espindola, Esther [Instituto Politecnico Nacional, Escuela Nacional de Ciencias Biologicas, Prolongacion Manuel M. Carpio s/n, esq. Plan de Ayala (Mexico)

    2011-06-15

    Colloidal silver nanoparticles were synthesized by reducing silver nitrate solutions with glucose, in the presence of gelatin as capping agent. The obtained nanoparticles were characterized by means of UV-Vis spectroscopy, transmission electron microscopy (TEM), and Fourier transform infrared (FTIR) spectroscopy. The response surface methodology (RSM) was also used to determine the influence of the variables on the size of the nanoparticles. The antifungal activity of the silver nanoparticles was evaluated on the phytopathogen Colletotrichum gloesporioides, which causes anthracnose in a wide range of fruits. The UV-Vis spectra indicated the formation of silver nanoparticles preferably spherical and of relatively small size (<20 nm). The above-mentioned was confirmed by TEM, observing a size distribution of 5-24 nm. According to RSM the synthesis variables influenced on the size of the silver nanoparticles. By means of FTIR spectroscopy it was determined that gelatin, through their amide and hydroxyl groups, interacts with nanoparticles preventing their agglomeration. The growth of C. gloesporioides in the presence of silver nanoparticles was significantly delayed in a dose dependent manner.

  5. OPTIMIZATION AND EVALUATION OF MICROBE FORTIFIED COMPOSTS AS BIOCONTROL AGENTS AGAINST PHYTOPATHOGENIC FUNGI

    Directory of Open Access Journals (Sweden)

    Ajinath S. Dukare

    2013-04-01

    Full Text Available A set of bacterial (B1-10 and cyanobacterial strains (C1-C14 were evaluated for their fungicidal activity against selected phytopathogenic fungi - Fusarium solani, Fusarium oxysporum, Fusarium oxysporum lycopersici, Fusarium moniliforme, Pythium debaryanum and Rhizoctonia solani. Further, they were used to amend rice straw compost and the filtrates were evaluated against the selected fungi after 10 and 20 days of incubation. Six promising strains, including three bacterial and three cyanobacterial strains were selected and characterised in terms of activity of hydrolytic enzymes. Interestingly, C12 strain (Anabaena spp. showed highest activity of cellulase, chitosanase and β 1, 3 glucanase. These strains were then evaluated by optimization of inoculum levels (1-5% in the rice straw compost. The strains B3, B5, C8 and C12 were observed to be most promising as they exhibited inhibition and significantly higher activity of microbiological parameters and hydrolytic enzymes at 1-2% level of inoculum in the compost. Further investigations are being undertaken to scale up the development of compost based biocontrol agents using these strains for evaluation at field level.

  6. Differences in stability of seed-associated microbial assemblages in response to invasion by phytopathogenic microorganisms

    Directory of Open Access Journals (Sweden)

    Samir Rezki

    2016-04-01

    Full Text Available Seeds are involved in the vertical transmission of microorganisms from one plant generation to another and consequently act as reservoirs for the plant microbiota. However, little is known about the structure of seed-associated microbial assemblages and the regulators of assemblage structure. In this work, we have assessed the response of seed-associated microbial assemblages of Raphanus sativus to invading phytopathogenic agents, the bacterial strain Xanthomonas campestris pv. campestris (Xcc 8004 and the fungal strain Alternaria brassicicola Abra43. According to the indicators of bacterial (16S rRNA gene and gyrB sequences and fungal (ITS1 diversity employed in this study, seed transmission of the bacterial strain Xcc 8004 did not change the overall composition of resident microbial assemblages. In contrast seed transmission of Abra43 strongly modified the richness and structure of fungal assemblages without affecting bacterial assemblages. The sensitivity of seed-associated fungal assemblage to Abra43 is mostly related to changes in relative abundance of closely related fungal species that belong to the Alternaria genus. Variation in stability of the seed microbiota in response to Xcc and Abra43 invasions could be explained by differences in seed transmission pathways employed by these micro-organisms, which ultimately results in divergence in spatio-temporal colonization of the seed habitat.

  7. Control of the phytopathogen Botrytis cinerea using adipic acid monoethyl ester.

    Science.gov (United States)

    Vicedo, Begonya; de la O Leyva, María; Flors, Víctor; Finiti, Ivan; Del Amo, Gemma; Walters, Dale; Real, Maria Dolores; García-Agustín, Pilar; González-Bosch, Carmen

    2006-01-01

    The in vitro and in vivo antifungal activity of adipic acid monoethyl ester (AAME) on the necrotrophic pathogen Botrytis cinerea has been studied. This chemical effectively controlled this important phytopathogen, inhibited spore germination and mycelium development at non-phytotoxic concentrations. The effectiveness of AAME treatment is concentration-dependent and influenced by pH. Spore germination in the presence of AAME is stopped at a very early stage, preventing germ tube development. In addition, cytological changes such as retraction of the conidial cytoplasm in the fungus are observed. AAME was also found to act on membrane integrity, affecting permeability without exhibiting lytic activity, as described previously for other antifungal compounds. Polyamine content in the mycelium of B. cinerea was also affected in response to AAME treatment, resulting in putrescine reduction and spermine accumulation similar to a number of antifungal agents. Microscopic observation of treated conidia after inoculation on tomato leaves suggested that inhibited spores are not able to attach to and penetrate the leaf. Finally, AAME completely suppressed the grey mould disease of tomato fruits under controlled inoculation conditions, providing evidence for its efficacy in a biological context and for the potential use of this chemical as an alternative fungicide treatment.

  8. The effects of some polyamine biosynthetic inhibitors on growth and morphology of phytopathogenic fungi

    Science.gov (United States)

    Rajam, M. V.; Galston, A. W.

    1985-01-01

    We have studied the effects of two polyamine biosynthetic inhibitors, alpha-difluoromethylornithine (DFMO) and alpha-difluoromethylarginine (DFMA), and of polyamines (PAs), alone and in combination, on mycelial growth and morphology of four phytopathogenic fungi: Botrytis sp, B. cinerea, Rhizoctonia solani and Monilinia fructicola. The inhibitors were added to a Czapek agar medium to get final concentrations of 0.1, 0.5 and 1.0 mM. DFMO and DFMA, suicide inhibitors of ornithine decarboxylase (ODC) and arginine decarboxylase (ADC) respectively, inhibited mycelial growth strongly; the effect was generally more pronounced with DFMA than with DFMO, but each fungus had its own response pattern. The addition of the PAs putrescine (Put) and spermidine (Spd) to the culture medium resulted in a promotion of growth. In Botrytis sp and Monilinia fructicola exposed to inhibitors plus PAs, mycelial growth was actually increased above control values. Mycelial morphology was altered and cell size dramatically reduced in plates containing inhibitors alone, whereas with PAs alone, or in combination with inhibitors, morphology was normal, but cell length and diameters increased considerably. These results suggest that PAs are essential for growth in fungal mycelia. The inhibition caused by DFMA may be due to its arginase-mediated conversion to DFMO.

  9. Efeito de herbicidas sobre agentes fitopatogênicos = Effect of herbicides on phytopathogenic agents

    Directory of Open Access Journals (Sweden)

    Daniel Dias Rosa

    2010-07-01

    Full Text Available Na agricultura moderna, diversas tecnologias auxiliam no aumento daprodutividade, sendo o herbicida uma delas, mas existem consequências atreladas ao seu uso, como os diversos efeitos sobre organismos não alvos. Neste trabalho, objetivou-se verificar esses efeitos sobre agentes fitopatogênicos, assim como avaliar o efeito do herbicida glyphosate sobre diversas doenças, em plantas de soja transgênicas.Verificou-se forte ação fungicida com o uso do herbicida glyphosate, assim como os outros avaliados “in vitro”, sobre os fungos testados, e os mesmos resultados foram observados nas plantas em condição de campo.In modern agriculture, several technologies have helped increase productivity, and herbicide is one of them. However, there are consequences linked to its use, such as the various effects on non-target organisms. The purpose of this work was to verify these effects on phytopathogenic agents, as well as assess the effect of glyphosate on diseases in transgenic soybean. There was a strong fungicide action using glyphosate herbicide as well as with the others evaluated in vitro regarding fungi tested. The same results were observed in plants in field conditions.

  10. Characterization of a novel botybirnavirus isolated from a phytopathogenic Alternaria fungus.

    Science.gov (United States)

    Xiang, Jun; Fu, Min; Hong, Ni; Zhai, Lifeng; Xiao, Feng; Wang, Guoping

    2017-12-01

    Alternaria fungi are important pathogens infecting a wide variety of organisms. Here, we report a novel double-stranded RNA (dsRNA) mycovirus named Alternaria botybirnavirus 1 (ABRV1) isolated from a phytopathogenic Alternaria sp. strain (SCFS-3) infecting a pear tree in China. ABRV1 has two dsRNA components (dsRNAs 1 and 2) with the sizes of 6,188 and 5,903 bp, containing two putative open reading frames encoding two polyproteins (202 and 192 kDa, respectively). The polyprotein encoded by ABRV1 dsRNA1 shares 41% amino acid (aa) sequence identity with the one encoded by dsRNA2 (instead of dsRNA1) of Sclerotinia sclerotiorum botybirnavirus 1 (SsBRV1). Conversely, the polyprotein encoded by ABRV1 dsRNA2 shares 46% aa sequence identity with the one (i.e., cap-pol fusion protein) encoded by SsBRV1 dsRNA1. ABRV1 has isometric spherical virus particles (~40 nm in diameter), putatively composed of the 60-, 70- and 80-kDa structural proteins. The genomic organization and phylogenetic analyses revealed that ABRV1 belongs to a newly proposed family "Botybirnaviridae", and to our knowledge, this is the first report of a botybirnavirus infecting an Alternaria sp. strain.

  11. Genome sequence of a novel mitovirus identified in the phytopathogenic fungus Alternaria arborescens.

    Science.gov (United States)

    Komatsu, Ken; Katayama, Yukie; Omatsu, Tsutomu; Mizutani, Tetsuya; Fukuhara, Toshiyuki; Kodama, Motoichiro; Arie, Tsutomu; Teraoka, Tohru; Moriyama, Hiromitsu

    2016-09-01

    The phytopathogenic fungus Alternaria spp. contains a variety of double-stranded RNA (dsRNA) elements of different sizes. Detailed analysis of next-generation sequencing data obtained using dsRNA purified from Alternaria arborescens, from which we had previously found Alternaria arborescens victorivirus 1, revealed the presence of another mycoviral-like dsRNA of approximately 2.5 kbp in length. When using the fungal mitochondrial genetic code, this dsRNA has a single open reading frame that potentially encodes an RNA-dependent RNA polymerase (RdRp) with significant to sequence similarity to those of viruses of the genus Mitovirus. Moreover, both the 5'- and 3'-untranslated regions have the potential to fold into stable stem-loop structures, which is characteristic of mitoviruses. Pairwise comparisons and phylogenetic analysis of the deduced amino acid sequences of RdRp indicated that the virus we identified in A. arborescens is a distinct member of the genus Mitovirus in the family Narnaviridae, designated as "Alternaria arborescens mitovirus 1" (AaMV1).

  12. Isolation of Bacteria with Antifungal Activity against the Phytopathogenic Fungi Stenocarpella maydis and Stenocarpella macrospora

    Directory of Open Access Journals (Sweden)

    Yuridia Mercado-Flores

    2011-08-01

    Full Text Available Stenocarpella maydis and Stenocarpella macrospora are the causal agents of ear rot in corn, which is one of the most destructive diseases in this crop worldwide. These fungi are important mycotoxin producers that cause different pathologies in farmed animals and represent an important risk for humans. In this work, 160 strains were isolated from soil of corn crops of which 10 showed antifungal activity against these phytopathogens, which, were identified as: Bacillus subtilis, Pseudomonas spp., Pseudomonas fluorescens, and Pantoea agglomerans by sequencing of 16S rRNA gene and the phylogenetic analysis. From cultures of each strain, extracellular filtrates were obtained and assayed to determine antifungal activity. The best filtrates were obtained in the stationary phase of B. subtilis cultures that were stable to the temperature and extreme pH values; in addition they did not show a cytotoxicity effect against brine shrimp and inhibited germination of conidia. The bacteria described in this work have the potential to be used in the control of white ear rot disease.

  13. Transmission of Fusarium boothii mycovirus via protoplast fusion causes hypovirulence in other phytopathogenic fungi.

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    Kyung-Mi Lee

    Full Text Available There is increasing concern regarding the use of fungicides to control plant diseases, whereby interest has increased in the biological control of phytopathogenic fungi by the application of hypovirulent mycoviruses as a possible alternative to fungicides. Transmission of hypovirulence-associated double-stranded RNA (dsRNA viruses between mycelia, however, is prevented by the vegetative incompatibility barrier that often exists between different species or strains of filamentous fungi. We determined whether protoplast fusion could be used to transmit FgV1-DK21 virus, which is associated with hypovirulence on F. boothii (formerly F. graminearum strain DK21, to F. graminearum, F. asiaticum, F. oxysporum f. sp. lycopersici, and Cryphonectria parasitica. Relative to virus-free strains, the FgV1-DK21 recipient strains had reduced growth rates, altered pigmentation, and reduced virulence. These results indicate that protoplast fusion can be used to introduce FgV1-DK21 dsRNA into other Fusarium species and into C. parasitica and that FgV1-DK21 can be used as a hypovirulence factor and thus as a biological control agent.

  14. Entomopathogenic Fungi as Dual Control Agents against Both the Pest Myzus persicae and Phytopathogen Botrytis cinerea.

    Science.gov (United States)

    Yun, Hwi-Geon; Kim, Dong-Jun; Gwak, Won-Seok; Shin, Tae-Young; Woo, Soo-Dong

    2017-09-01

    The green peach aphid ( Myzus persicae ), a plant pest, and gray mold disease, caused by Botrytis cinerea , affect vegetables and fruit crops all over the world. To control this aphid and mold, farmers typically rely on the use of chemical insecticides or fungicides. However, intensive use of these chemicals over many years has led to the development of resistance. To overcome this problem, there is a need to develop alternative control methods to suppress populations of this plant pest and pathogen. Recently, potential roles have been demonstrated for entomopathogenic fungi in endophytism, phytopathogen antagonism, plant growth promotion, and rhizosphere colonization. Here, the antifungal activities of selected fungi with high virulence against green peach aphids were tested to explore their potential for the dual control of B. cinerea and M. persicae . Antifungal activities against B. cinerea were evaluated by dual culture assays using both aerial conidia and cultural filtrates of entomopathogenic fungi. Two fungal isolates, Beauveria bassiana SD15 and Metarhizium anisopliae SD3, were identified as having both virulence against aphids and antifungal activity. The virulence of these isolates against aphids was further tested using cultural filtrates, blastospores, and aerial conidia. The most virulence was observed in the simultaneous treatment with blastospores and cultural filtrate. These results suggest that the two fungal isolates selected in this study could be used effectively for the dual control of green peach aphids and gray mold for crop protection.

  15. A putative twin-arginine translocation system in the phytopathogenic bacterium Xylella fastidiosa.

    Science.gov (United States)

    Ciapina, Luciane Prioli; Picchi, Simone Cristina; Lacroix, Jean-Marie; Lemos, Eliana Gertrudes de Macedo; Ödberg-Ferragut, Carmen

    2011-02-01

    The twin-arginine translocation (Tat) pathway of the xylem-limited phytopathogenic bacterium Xylella fastidiosa strain 9a5c, responsible for citrus variegated chlorosis, was explored. The presence of tatA, tatB, and tatC in the X. fastidiosa genome together with a list of proteins harboring 2 consecutive arginines in their signal peptides suggested the presence of a Tat pathway. The functional Tat dependence of X. fastidiosa OpgD was examined. Native or mutated signal peptides were fused to the β-lactamase. Expression of fusion with intact signal peptides mediated high resistance to ampicillin in Escherichia coli tat+ but not in the E. coli tat null mutant. The replacement of the 2 arginines by 2 lysines prevented the export of β-lactamase in E. coli tat+, demonstrating that X. fastidiosa OpgD carries a signal peptide capable of engaging the E. coli Tat machinery. RT-PCR analysis revealed that the tat genes are transcribed as a single operon. tatA, tatB, and tatC genes were cloned. Complementation assays in E. coli devoid of all Tat or TatC components were unsuccessful, whereas X. fastidiosa Tat components led to a functional Tat translocase in E. coli TatB-deficient strain. Additional experiments implicated that X. fastidiosa TatB component could form a functional heterologous complex with the E. coli TatC component.

  16. Zoosporicidal activity of polyflavonoid tannin identified in Lannea coromandelica stem bark against phytopathogenic oomycete Aphanomyces cochlioides.

    Science.gov (United States)

    Islam, Md Tofazzal; Ito, Toshiaki; Sakasai, Mitsuyoshi; Tahara, Satoshi

    2002-11-06

    In a survey of nonhost plant secondary metabolites regulating motility and viability of zoospores of the Aphanomyces cochlioides, we found that stem bark extracts of Lannea coromandelica remarkably inhibited motility of zoospores followed by lysis. Bioassay-guided fractionation and chemical characterization of Lannea extracts by MALDI-TOF-MS revealed that the active constituents were angular type polyflavonoid tannins. Commercial polyflavonoid tannins, Quebracho and Mimosa, also showed identical zoosporicidal activity. Against zoospores, the motility-inhibiting and lytic activities were more pronounced in Lannea extracts (MIC 0.1 microg/mL) than in Quebracho (MIC 0.5 microg/mL) and Mimosa (MIC 0.5 microg/mL). Scanning electron microscopic observation visualized that both Lannea and commercial tannins caused lysis of cell membrane followed by fragmentation of cellular materials. Naturally occurring polyflavonoid tannin merits further study as potential zoospore regulating agent or as lead compound. To the best of our knowledge, this is the first report of zoosporicidal activity of natural polyflavonoid tannins against an oomycete phytopathogen.

  17. Screening for Antimicrobial Activity of Wood Rotting Higher Basidiomycetes Mushrooms from Uruguay against Phytopathogens.

    Science.gov (United States)

    Barneche, Stephanie; Jorcin, Gabriela; Cecchetto, Gianna; Cerdeiras, María Pía; Vázquez, Alvaro; Alborés, Silvana

    2016-01-01

    In this work, the antimicrobial activity of extracts of wood rotting higher Basidiomycetes mushrooms isolated from Eucalyptus plantations in Uruguay was studied using bacterial and fungal phytopathogens as targets. Fifty-one extracts from mycelia and growth broth were prepared from higher Basidiomycetes mushrooms, from which eight extracts (from Ganoderma resinaceum, Laetiporus sulphureus, Dictyopanus pusillus, and Bjerkandera adusta) showed antimicrobial activity against Xanthomonas vesicatoria, Aspergillus oryzae, Penicillium expansum, Botrytis cinerea, and Rhizopus stolonifer as assayed in the qualitative test. The minimum inhibitory concentration (MIC) for those fungal extracts was determined and the results showed that L. sulphureus deserved further study, with low MIC values against X. vesicatoria. The antimicrobial activity of L. sulphureus culture broth extracts grown under different culture conditions was evaluated against X. vesicatoria. From the results of these assays, larger-scale cultures for the production of the compound(s) with antimicrobial activity should be performed using malt extract broth, at pH 5, at 20°C and static culture conditions.

  18. Activity and mode of action against fungal phytopathogens of bovine lactoferricin-derived peptides.

    Science.gov (United States)

    Muñoz, A; Marcos, J F

    2006-12-01

    To evaluate the activity against fungal phytopathogens of two synthetic peptides derived from the protein bovine lactoferricin: the antibacterial active core of six amino acid residues (LfcinB(20-25)) and an extension of 15 amino acids (LfcinB(17-31)). In vitro activity against fungal pathogens was determined and compared with that against model micro-organisms. Activity was demonstrated against fungi of agronomic relevance. Distinct antimicrobial properties in vitro were found for the two peptides. LfcinB(17-31) had growth inhibitory activity higher than LfcinB(20-25). However, LfcinB(17-31) was not fungicidal to quiescent conidia of Penicillium digitatum at the concentrations assayed, while LfcinB(20-25) killed conidia more efficiently. Microscopical observations showed that the mycelium of P. digitatum treated with LfcinB(17-31) developed alterations of growth, sporulation and chitin deposition, and permeation of hyphal cells. In experimental inoculations of mandarins, both peptides showed limited protective effect against the disease caused by P. digitatum. LfcinB(20-25) and LfcinB(17-31) peptides were shown to have antimicrobial activity against plant pathogenic filamentous fungi, with distinct properties and mode of action. LfcinB(20-25) and LfcinB(17-31) peptides offer novel alternatives to develop resistant plants by molecular breeding.

  19. Asparaginase Erwinia chrysanthemi as a component of a multi-agent chemotherapeutic regimen for the treatment of patients with acute lymphoblastic leukemia who have developed hypersensitivity to E. coli-derived asparaginase.

    Science.gov (United States)

    Figueiredo, Lisa; Cole, Peter D; Drachtman, Richard A

    2016-03-01

    Asparaginase has been a mainstay of therapy in the treatment of acute lymphoblastic leukemia since the 1970s. There are two major preparations available and FDA approved in the United States today, one derived from Escherichia coli and the other from Erwinia chrysanthemi. Erwinia asparaginase is antigenically distinct from and has a considerably shorter biological half-life than E coli asparaginase. Erwinia asparaginase has been used in cases of hypersensitivity to E. coli-derived asparaginases, which has been reported in up to 30% of patients. While PEG asparaginase is increasingly used in front-line therapy for ALL, hypersensitivity still occurs with this preparation, and a change to a non-cross-reactive preparation may be necessary.

  20. 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

  1. Seca de ponteiros causada por Erwinia psidii: diversidade genética do patógeno e resistência em Eucalyptus spp.

    OpenAIRE

    Hermenegildo, Pollyane da Silva

    2015-01-01

    A seca dos ponteiros causada por Erwinia psidii é um dos fatores limitantes para a produção de goiaba no Brasil. Recentemente, essa bactéria foi relatada como o agente causal da seca de ponteiros e murcha de Eucalyptus spp. no Brasil, Uruguai e Argentina. Como se trata de uma enfermidade recentemente descoberta, há ainda poucos estudos sobre o patossistema E. psidii - eucalipto, especialmente quanto à variabilidade genética do patógeno e resistência de Eucalyptus à doença. Assim, neste trabal...

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

    Science.gov (United States)

    Pester, Doris; Milčevičová, Renáta; Schaffer, Johann; Wilhelm, Eva; Blümel, Sylvia

    2012-01-01

    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. 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). 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 control on a molecular level.

  3. A complete structural characterization of the desferrioxamine E biosynthetic pathway from the fire blight pathogen Erwinia amylovora.

    Science.gov (United States)

    Salomone-Stagni, Marco; Bartho, Joseph D; Polsinelli, Ivan; Bellini, Dom; Walsh, Martin A; Demitri, Nicola; Benini, Stefano

    2018-02-08

    The Gram-negative bacterium Erwinia amylovora is the etiological agent of fire blight, a devastating disease which affects Rosaceae such as apple, pear and quince. The siderophore desferrioxamine E plays an important role in bacterial pathogenesis by scavenging iron from the host. DfoJ, DfoA and DfoC are the enzymes responsible for desferrioxamine production starting from lysine. We have determined the crystal structures of each enzyme in the desferrioxamine E pathway and demonstrate that the biosynthesis involves the concerted action of DfoJ, followed by DfoA and lastly DfoC. These data provide the first crystal structures of a Group II pyridoxal-dependent lysine decarboxylase, a cadaverine monooxygenase and a desferrioxamine synthetase. DfoJ is a homodimer made up of three domains. Each monomer contributes to the completion of the active site, which is positioned at the dimer interface. DfoA is the first structure of a cadaverine monooxygenase. It forms homotetramers whose subunits are built by two domains: one for FAD and one for NADP + binding, the latter of which is formed by two subdomains. We propose a model for substrate binding and the role of residues 43-47 as gate keepers for FAD binding and the role of Arg97 in cofactors turnover. DfoC is the first structure of a desferrioxamine synthetase and the first of a multi-enzyme siderophore synthetase coupling an acyltransferase domain with a Non-Ribosomal Peptide Synthetase (NRPS)-Independent Siderophore domain (NIS). Copyright © 2018 Elsevier Inc. All rights reserved.

  4. 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.

  5. CEREALS ASSESSMENT TOWARDS CONTAMINATION OF PHYTOPATHOGENIC FUNGI IN FOREST-STEPPE AREA OF UKRAINE

    Directory of Open Access Journals (Sweden)

    V. B. Yekimova

    2014-11-01

    Full Text Available The usage of high-quality seed, not affected by pathogens and fungi promote the high and stable yields. The condition of seeds determines their germination, seedling damage and adult plants, which ultimately affects the yield and on their quality. The significant risk of seed infection was registered in years with high humidity in the pre-harvest and harvest period. Therefore, along with the traditional seed control it is necessary to conduct phytopahtology expertise, allowing to identify the species composition of the microflora of seeds and the degree of infection with different pathogens. Smut disease - a solid and loose smut of wheat, hard and loose smut of barley - cause crop losses are clear - in the form of the destruction of the ear, and hidden - in the form of a reduction in seed germination, reduce winter hardiness, plant growth inhibition. Pathogens root rot causing blight, Fusarium and Helminthosporium that insignificantly demand on environmental conditions and extremely plastic. They have large set of enzymes that can exist on a variety of substrates, and therefore are widely distributed in nature and cause considerable damage to crops. Especially significant losses occur if the humidity during the ripening grain observed for several seasons, which leads to accumulation of the infection naturally. The aim of research was to assess the contamination of grain of spring wheat and barley phytopathogenic fungi in one of the main grain regions of Ukraine - the southern forest. The research conducted during the summer 2014 and compared with the results of previous years. Samples were taken from different plots. Seed contamination of samples by various phytopathogenic fungi was determined by the number of infected kernels per 100 seed sample. Frequency of registered species was recorded. For every studied sample we set the percentage of species. Analysis of the grain on the fungal infection and avdelenie in pure culture was performed

  6. CEREALS ASSESSMENT TOWARDS CONTAMINATION OF PHYTOPATHOGENIC FUNGI IN FOREST-STEPPE AREA OF UKRAINE

    Directory of Open Access Journals (Sweden)

    Yekimova V. B.

    2014-12-01

    Full Text Available The usage of high-quality seed, not affected by pathogens and fungi promote the high and stable yields. The condition of seeds determines their germination, seedling damage and adult plants, which ultimately affects the yield and on their quality. The significant risk of seed infection was registered in years with high humidity in the pre-harvest and harvest period. Therefore, along with the traditional seed control it is necessary to conduct phytopahtology expertise, allowing to identify the species composition of the microflora of seeds and the degree of infection with different pathogens. Smut disease - a solid and loose smut of wheat, hard and loose smut of barley - cause crop losses are clear - in the form of the destruction of the ear, and hidden - in the form of a reduction in seed germination, reduce winter hardiness, plant growth inhibition. Pathogens root rot causing blight, Fusarium and Helminthosporium that insignificantly demand on environmental conditions and extremely plastic. They have large set of enzymes that can exist on a variety of substrates, and therefore are widely distributed in nature and cause considerable damage to crops. Especially significant losses occur if the humidity during the ripening grain observed for several seasons, which leads to accumulation of the infection naturally. The aim of research was to assess the contamination of grain of spring wheat and barley phytopathogenic fungi in one of the main grain regions of Ukraine - the southern forest. The research conducted during the summer 2014 and compared with the results of previous years. Samples were taken from different plots. Seed contamination of samples by various phytopathogenic fungi was determined by the number of infected kernels per 100 seed sample. Frequency of registered species was recorded. For every studied sample we set the percentage of species. Analysis of the grain on the fungal infection and avdelenie in pure culture was performed

  7. Hypovirulence of the phytopathogenic fungus Botryosphaeria dothidea: association with a coinfecting chrysovirus and a partitivirus.

    Science.gov (United States)

    Wang, LiPing; Jiang, JingJing; Wang, YanFen; Hong, Ni; Zhang, Fangpeng; Xu, WenXing; Wang, GuoPing

    2014-07-01

    Botryosphaeria dothidea is an important pathogenic fungus causing fruit rot, leaf and stem ring spots and dieback, stem canker, stem death or stool mortality, and decline of pear trees. Seven double-stranded RNAs (dsRNAs; dsRNAs 1 to 7 with sizes of 3,654, 2,773, 2,597, 2,574, 1,823, 1,623, and 511 bp, respectively) were identified in an isolate of B. dothidea exhibiting attenuated growth and virulence and a sectoring phenotype. Characterization of the dsRNAs revealed that they belong to two dsRNA mycoviruses. The four largest dsRNAs (dsRNAs 1 to 4) are the genomic components of a novel member of the family Chrysoviridae (tentatively designated Botryosphaeria dothidea chrysovirus 1 [BdCV1]), a view supported by the morphology of the virions and phylogenetic analysis of the putative RNA-dependent RNA polymerases (RdRps). Two other dsRNAs (dsRNAs 5 and 6) are the genomic components of a novel member of the family Partitiviridae (tentatively designated Botryosphaeria dothidea partitivirus 1 [BdPV1]), which is placed in a clade distinct from other established partitivirus genera on the basis of the phylogenetic analysis of its RdRp. The smallest dsRNA, dsRNA7, seems to be a noncoding satellite RNA of BdPV1 on the basis of the conservation of its terminal sequences in BdPV1 genomic segments and its cosegregation with BdPV1 after horizontal transmission. This is the first report of a chrysovirus and a partitivirus infecting B. dothidea and of a chrysovirus associated with the hypovirulence of a phytopathogenic fungus. Our studies identified and characterized two novel mycoviruses, Botryosphaeria dothidea chrysovirus 1 (BdCV1) and Botryosphaeria dothidea partitivirus 1 (BdPV1), associated with the hypovirulence of an important fungus pathogenic to fruit trees. This is the first report of a chrysovirus and a partitivirus infecting B. dothidea and of a chrysovirus associated with the hypovirulence of a phytopathogenic fungus. BdCV1 appears to be a good candidate for the

  8. Impact of phytopathogen infection and extreme weather stress on internalization of Salmonella Typhimurium in lettuce.

    Science.gov (United States)

    Ge, Chongtao; Lee, Cheonghoon; Nangle, Ed; Li, Jianrong; Gardner, David; Kleinhenz, Matthew; Lee, Jiyoung

    2014-01-03

    Internalization of human pathogens, common in many types of fresh produce, is a threat to human health since the internalized pathogens cannot be fully inactivated/removed by washing with water or sanitizers. Given that pathogen internalization can be affected by many environmental factors, this study was conducted to investigate the influence of two types of plant stress on the internalization of Salmonella Typhimurium in iceberg lettuce during pre-harvest. The stresses were: abiotic (water stress induced by extreme weather events) and biotic (phytopathogen infection by lettuce mosaic virus [LMV]). Lettuce with and without LMV infection were purposefully contaminated with green fluorescence protein-labeled S. Typhimurium on the leaf surfaces. Lettuce was also subjected to water stress conditions (drought and storm) which were simulated by irrigating with different amounts of water. The internalized S. Typhimurium in the different parts of the lettuce were quantified by plate count and real-time quantitative PCR and confirmed with a laser scanning confocal microscope. Salmonella internalization occurred under the conditions outlined above; however internalization levels were not significantly affected by water stress alone. In contrast, the extent of culturable S. Typhimurium internalized in the leafy part of the lettuce decreased when infected with LMV under water stress conditions and contaminated with high levels of S. Typhimurium. On the other hand, LMV-infected lettuce showed a significant increase in the levels of culturable bacteria in the roots. In conclusion, internalization was observed under all experimental conditions when the lettuce surface was contaminated with S. Typhimurium. However, the extent of internalization was only affected by water stress when lettuce was infected with LMV. © 2013.

  9. Leaf-cutting ant fungi produce cell wall degrading pectinase complexes reminiscent of phytopathogenic fungi

    Directory of Open Access Journals (Sweden)

    Boomsma Jacobus J

    2010-12-01

    those normally found in phytopathogens.

  10. Leaf-cutting ant fungi produce cell wall degrading pectinase complexes reminiscent of phytopathogenic fungi.

    Science.gov (United States)

    Schiøtt, Morten; Rogowska-Wrzesinska, Adelina; Roepstorff, Peter; Boomsma, Jacobus J

    2010-12-31

    Leaf-cutting (attine) ants use their own fecal material to manure fungus gardens, which consist of leaf material overgrown by hyphal threads of the basidiomycete fungus Leucocoprinus gongylophorus that lives in symbiosis with the ants. Previous studies have suggested that the fecal droplets contain proteins that are produced by the fungal symbiont to pass unharmed through the digestive system of the ants, so they can enhance new fungus garden growth. We tested this hypothesis by using proteomics methods to determine the gene sequences of fecal proteins in Acromyrmex echinatior leaf-cutting ants. Seven (21%) of the 33 identified proteins were pectinolytic enzymes that originated from the fungal symbiont and which were still active in the fecal droplets produced by the ants. We show that these enzymes are found in the fecal material only when the ants had access to fungus garden food, and we used quantitative polymerase chain reaction analysis to show that the expression of six of these enzyme genes was substantially upregulated in the fungal gongylidia. These unique structures serve as food for the ants and are produced only by the evolutionarily advanced garden symbionts of higher attine ants, but not by the fungi reared by the basal lineages of this ant clade. Pectinolytic enzymes produced in the gongylidia of the fungal symbiont are ingested but not digested by Acromyrmex leaf-cutting ants so that they end up in the fecal fluid and become mixed with new garden substrate. Substantial quantities of pectinolytic enzymes are typically found in pathogenic fungi that attack live plant tissue, where they are known to breach the cell walls to allow the fungal mycelium access to the cell contents. As the leaf-cutting ant symbionts are derived from fungal clades that decompose dead plant material, our results suggest that their pectinolytic enzymes represent secondarily evolved adaptations that are convergent to those normally found in phytopathogens.

  11. Septins from the phytopathogenic fungus Ustilago maydis are required for proper morphogenesis but dispensable for virulence.

    Directory of Open Access Journals (Sweden)

    Isabel Alvarez-Tabarés

    Full Text Available BACKGROUND: Septins are a highly conserved family of GTP-binding proteins involved in multiple cellular functions, including cell division and morphogenesis. Studies of septins in fungal cells underpin a clear correlation between septin-based structures and fungal morphology, providing clues to understand the molecular frame behind the varied morphologies found in fungal world. METHODOLOGY/PRINCIPAL FINDINGS: Ustilago maydis genome has the ability to encode four septins. Here, using loss-of-function as well as GFP-tagged alleles of these septin genes, we investigated the roles of septins in the morphogenesis of this basidiomycete fungus. We described that septins in U. maydis could assemble into at least three different structures coexisting in the same cell: bud neck collars, band-like structures at the growing tip, and long septin fibers that run from pole to pole near the cell cortex. We also found that in the absence of septins, U. maydis cells lost their elongated shape, became wider at the central region and ended up losing their polarity, pointing to an important role of septins in the morphogenesis of this fungus. These morphological defects were alleviated in the presence of an osmotic stabilizer suggesting that absence of septins affected the proper formation of the cell wall, which was coherent with a higher sensitivity of septin defective cells to drugs that affect cell wall construction as well as exocytosis. As U. maydis is a phytopathogen, we analyzed the role of septins in virulence and found that in spite of the described morphological defects, septin mutants were virulent in corn plants. CONCLUSIONS/SIGNIFICANCE: Our results indicated a major role of septins in morphogenesis in U. maydis. However, in contrast to studies in other fungal pathogens, in which septins were reported to be necessary during the infection process, we found a minor role of septins during corn infection by U. maydis.

  12. Ag doped hollow TiO2 nanoparticles as an effective green fungicide against Fusarium solani and Venturia inaequalis phytopathogens

    International Nuclear Information System (INIS)

    Boxi, Siddhartha Sankar; Mukherjee, Khushi; Paria, Santanu

    2016-01-01

    Chemical-based pesticides are widely used in agriculture to protect crops from insect infestation and diseases. However, the excessive use of highly toxic pesticides causes several human health (neurological, tumor, cancer) and environmental problems. Therefore nanoparticle-based green pesticides have become of special importance in recent years. The antifungal activities of pure and Ag doped (solid and hollow) TiO 2 nanoparticles are studied against two potent phytopathogens, Fusarium solani (which causes Fusarium wilt disease in potato, tomato, etc) and Venturia inaequalis (which causes apple scab disease) and it is found that hollow nanoparticles are more effective than the other two. The antifungal activities of the nanoparticles were further enhanced against these two phytopathogens under visible light exposure. The fungicidal effect of the nanoparticles depends on different parameters, such as particle concentration and the intensity of visible light. The minimum inhibitory dose of the nanoparticles for V. inaequalis and F. solani are 0.75 and 0.43 mg/plate. The presence of Ag as a dopant helps in the formation of stable Ag–S and disulfide bonds (R–S–S–R) in cellular protein, which leads to cell damage. During photocatalysis generated • OH radicals loosen the cell wall structure and this finally leads to cell death. The mechanisms of the fungicidal effect of nanoparticles against these two phytopathogens are supported by biuret and triphenyl tetrazolium chloride analyses and field emission electron microscopy. Apart from the fungicidal effect, at a very low dose (0.015 mg/plate) the nanoparticles are successful in arresting production of toxic napthoquinone pigment for F. solani which is related to the fungal pathogenecity. The nanoparticles are found to be effective in protecting potatoes affected by F. solani or other fungi from spoiling. (paper)

  13. Ag doped hollow TiO2 nanoparticles as an effective green fungicide against Fusarium solani and Venturia inaequalis phytopathogens

    Science.gov (United States)

    Sankar Boxi, Siddhartha; Mukherjee, Khushi; Paria, Santanu

    2016-02-01

    Chemical-based pesticides are widely used in agriculture to protect crops from insect infestation and diseases. However, the excessive use of highly toxic pesticides causes several human health (neurological, tumor, cancer) and environmental problems. Therefore nanoparticle-based green pesticides have become of special importance in recent years. The antifungal activities of pure and Ag doped (solid and hollow) TiO2 nanoparticles are studied against two potent phytopathogens, Fusarium solani (which causes Fusarium wilt disease in potato, tomato, etc) and Venturia inaequalis (which causes apple scab disease) and it is found that hollow nanoparticles are more effective than the other two. The antifungal activities of the nanoparticles were further enhanced against these two phytopathogens under visible light exposure. The fungicidal effect of the nanoparticles depends on different parameters, such as particle concentration and the intensity of visible light. The minimum inhibitory dose of the nanoparticles for V. inaequalis and F. solani are 0.75 and 0.43 mg/plate. The presence of Ag as a dopant helps in the formation of stable Ag-S and disulfide bonds (R-S-S-R) in cellular protein, which leads to cell damage. During photocatalysis generated •OH radicals loosen the cell wall structure and this finally leads to cell death. The mechanisms of the fungicidal effect of nanoparticles against these two phytopathogens are supported by biuret and triphenyl tetrazolium chloride analyses and field emission electron microscopy. Apart from the fungicidal effect, at a very low dose (0.015 mg/plate) the nanoparticles are successful in arresting production of toxic napthoquinone pigment for F. solani which is related to the fungal pathogenecity. The nanoparticles are found to be effective in protecting potatoes affected by F. solani or other fungi from spoiling.

  14. Draft genome sequence of Bacillus velezensis 2A-2B strain: a rhizospheric inhabitant of Sporobolus airoides (Torr.) Torr., with antifungal activity against root rot causing phytopathogens.

    Science.gov (United States)

    Martínez-Raudales, Inés; De La Cruz-Rodríguez, Yumiko; Alvarado-Gutiérrez, Alejandro; Vega-Arreguín, Julio; Fraire-Mayorga, Ahuitz; Alvarado-Rodríguez, Miguel; Balderas-Hernández, Victor; Fraire-Velázquez, Saúl

    2017-01-01

    A Bacillus velezensis strain from the rhizosphere of Sporobolus airoides (Torr.) Torr . , a grass in central-north México, was isolated during a biocontrol of phytopathogens scrutiny study. The 2A-2B strain exhibited at least 60% of growth inhibition of virulent isolates of phytopathogens causing root rot. These phytopathogens include Phytophthora capsici , Fusarium solani , Fusarium oxysporum and Rhizoctonia solani . Furthermore, the 2A-2B strain is an indolacetic acid producer, and a plant inducer of PR1, which is an induced systemic resistance related gene in chili pepper plantlets. Whole genome sequencing was performed to generate a draft genome assembly of 3.953 MB with 46.36% of GC content, and a N50 of 294,737. The genome contains 3713 protein coding genes and 89 RNA genes. Moreover, comparative genome analysis revealed that the 2A-2B strain had the greatest identity (98.4%) with Bacillus velezensis.

  15. Probing the phytopathogenic stem rot fungus with phytoalexins and analogues: unprecedented glucosylation of camalexin and 6-methoxycamalexin.

    Science.gov (United States)

    Pedras, M Soledade C; Ahiahonu, Pearson W K

    2002-10-01

    The remarkable metabolism of the cruciferous phytoalexins camalexin and 6-methoxycamalexin by the stem rot phytopathogen Sclerotinia sclerotiorum is reported. The biotransformations yielded camalexins glucosylated at N-1 or C-6 of the indole ring, with substantially lower antifungal activity than camalexins. A camalexin analogue with the positions N-1 and C-6 blocked was metabolized but at a much slower rate than the natural phytoalexins. The chemistry involved in the metabolism of natural camalexins and two new analogues, as well as their novel metabolites and respective antifungal activities is described.

  16. Derivation of Mutants of Erwinia carotovora subsp. betavasculorum Deficient in Export of Pectolytic Enzymes with Potential for Biological Control of Potato Soft Rot

    Science.gov (United States)

    Costa, José M.; Loper, Joyce E.

    1994-01-01

    Erwinia carotovora subsp. betavasculorum Ecb168 produces an antibiotic(s) that suppresses growth of the related bacterium Erwinia carotovora subsp. carotovora in culture and in wounds of potato tubers. Strain Ecb168 also produces and secretes pectolytic enzymes and causes a vascular necrosis and root rot of sugar beet. Genes (out) involved in secretion of pectolytic enzymes by Ecb168 were localized to two HindIII fragments (8.5 and 10.5 kb) of Ecb168 genomic DNA by hybridization to the cloned out region of E. carotovora subsp. carotovora and by complementation of Out- mutants of E. carotovora subsp. carotovora. Out- mutants of Ecb168, which did not secrete pectate lyase into the culture medium, were obtained when deletions internal to either HindIII fragment were introduced into the genome of Ecb168 through marker exchange mutagenesis. Out- mutants of Ecb168 were complemented to the Out+ phenotype by introduction of the corresponding cloned HindIII fragment. Out- mutants of Ecb168 were less virulent than the Out+ parental strain on potato tubers. Strain Ecb168 and Out- derivatives inhibited the growth of E. carotovora subsp. carotovora in culture, indicating that the uncharacterized antibiotic(s) responsible for antagonism was exported through an out-independent mechanism. Strain Ecb168 and Out- derivatives reduced the establishment of large populations of E. carotovora subsp. carotovora in wounds of potato tubers and suppressed tuber soft rot caused by E. carotovora subsp. carotovora. PMID:16349316

  17. 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.

  18. Inoculation of Malus genotypes with a set of Erwinia amylovora strains indicates a gene-for-gene relationship between the effector gene eop1 and both Malus floribunda 821 and Malus 'Evereste'

    Science.gov (United States)

    The Gram-negative bacterium Erwinia amylovora (Burrill) Winslow. et al., causal agent of fire blight disease in pome fruit trees, encodes a type three secretion system (T3SS) that functions to translocate effector proteins into plant cells that collectively function to suppress host defenses and ena...

  19. Whole-genome sequence of Pseudomonas fluorescens EK007-RG4, a promising biocontrol agent against a broad range of bacteria, including the fire blight bacterium Erwinia amylovora

    DEFF Research Database (Denmark)

    Habibi, Roghayeh; Tarighi, Saeed; Behravan, Javad

    2017-01-01

    Here, we report the first draft whole-genome sequence of Pseudomonas fluorescens strain EK007-RG4, which was isolated from the phylloplane of a pear tree. P. fluorescens EK007-RG4 displays strong antagonism against Erwinia amylovora, the causal agent for fire blight disease, in addition to several...

  20. Inhibitory effect of Thymus vulgaris and Origanum vulgare essential oils on virulence factors of phytopathogenic Pseudomonas syringae strains.

    Science.gov (United States)

    Carezzano, M E; Sotelo, J P; Primo, E; Reinoso, E B; Paletti Rovey, M F; Demo, M S; Giordano, W F; Oliva, M de Las M

    2017-07-01

    Pseudomonas syringae is a phytopathogenic bacterium that causes lesions in leaves during the colonisation process. The damage is associated with production of many virulence factors, such as biofilm and phytotoxins. The essential oils of Thymus vulgaris (thyme) and Origanum vulgare (oregano) have been demonstrated to inhibit P. syringae. The aim of this study was to investigate the effects of T. vulgaris and O. vulgare essential oils on production of virulence factors of phytopathogenic P. syringae strains, including anti-biofilm and anti-toxins activities. The broth microdilution method was used for determination of MIC and biofilm inhibition assays. Coronatine, syringomycin and tabtoxin were pheno- and genotypically evaluated. Both oils showed good inhibitory activity against P. syringae, with MIC values from 1.43 to 11.5 mg·ml -1 for thyme and 5.8 to 11.6 mg·ml -1 for oregano. Biofilm formation, production of coronatine, syringomycin and tabtoxin were inhibited by thyme and oregano essential oil in most strains. The results presented here are promising, demonstrating the bactericidal activity and reduction of virulence factor production after treatment with thyme and oregano oil, providing insight into how they exert their antibacterial activity. These natural products could be considered in the future for the control of diseases caused by P. syringae. © 2017 German Botanical Society and The Royal Botanical Society of the Netherlands.

  1. Selection of endophytic fungi from comfrey (Symphytum officinale L.) for in vitro biological control of the phytopathogen Sclerotinia sclerotiorum (Lib.)

    Science.gov (United States)

    Rocha, Rafaeli; da Luz, Daniela Eleutério; Engels, Cibelle; Pileggi, Sônia Alvim Veiga; de Souza Jaccoud Filho, David; Matiello, Rodrigo Rodrigues; Pileggi, Marcos

    2009-01-01

    Biological control consists of using one organism to attack another that may cause economic damage to crops. Integrated Pest Management (IPM) is a very common strategy. The white mold produced by Sclerotinia sclerotiorum (Lib.) causes considerable damage to bean crops. This fungus is a soil inhabitant, the symptoms of which are characterized by water-soaked lesions covered by a white cottony fungal growth on the soil surface and/or the host plant. Possible biological control agents taken from plants are being investigated as phytopathogen inhibitors. These are endophytic microorganisms that inhabit the intercellular spaces of vegetal tissues and are often responsible for antimicrobial production. The objective of the present study was to select endophytic fungi isolated from comfrey (Symphytum officinale L.) leaves with in vitro antagonist potential against the phytopathogenic fungus S. sclerotiorum. Twelve isolates of endophytic fungi and a pathogenic strain of S. sclerotiorum were used in the challenge method. With the aid of this method, four endophytes with the best antagonistic activity against S. sclerotiorum were selected. Pathogen growth inhibition zones were considered indicative of antibiosis. The percentages of pathogenic mycelia growth were measured both with and without the antagonist, resulting in growth reductions of 46.7% to 50.0% for S. sclerotiorum. These analyses were performed by evaluating the endophytic/pathogenic mycelia growth in mm/day over an eight-day period of antagonistic tests. PMID:24031320

  2. Potential antagonism of some Trichoderma strains isolated from Moroccan soil against three phytopathogenic fungi of great economic importance

    Directory of Open Access Journals (Sweden)

    Wafaa MOKHTARI

    2017-09-01

    Full Text Available In this study, 17 Trichoderma strains were isolated from different soils (crop fields and Argan forests in Morocco. Purified monospore cultures were identified using molecular methods and tested for their potential antagonism against three phytopathogenic fungi (Fusarium oxyxporum, verticillium dahlia and rhizoctonia solani. After DNA extraction, translation elongation factor (tef1 was amplified in extracts of 17 strains, sequenced and compared with their ex-types. As a result, three species were identified among the strains, which clustered in two different subclades of Trichoderma: the species T. afroharzianum, and T. guizhouense belong to the Harzianum clade, while T. longibrachiatum belongs to the Longibrachiatum clade. Investigation of potential antagonistic effects of these strains against the soil-borne phytopathogens F. oxysporum, R. solani and V. dahliae was conducted in a dual culture plate assay, using 17 promising Trichoderma strains that have been selected based on a polymerase chain reaction (PCR screening approach. In vitro, Trichoderma isolates showed effective antagonistic performance by decreasing soil borne pathogens mycelium radial growth. Trichoderma afroharzianum showed the highest Percentage of Radial Inhibition Growth (PRIG %. The highest PRIG% = 98% was for 8A2.3 isolate against R. solani and the lowest PRIG%= 67% for T9i10 against F. oxysporum. On the other hand, T9i12, which is T. reesei species, led to a high radial inhibition of pathogens’ mycelium.

  3. UVA Photoactivation of Harmol Enhances Its Antifungal Activity against the PhytopathogensPenicillium digitatumandBotrytis cinerea.

    Science.gov (United States)

    Olmedo, Gabriela M; Cerioni, Luciana; González, María M; Cabrerizo, Franco M; Volentini, Sabrina I; Rapisarda, Viviana A

    2017-01-01

    Phytopathogenic fungi responsible for post-harvest diseases on fruit and vegetables cause important economic losses. We have previously reported that harmol (1-methyl-9H-pyrido[3,4-b]indol-7-ol) is active against the causal agents of green and gray molds Penicillium digitatum and Botrytis cinerea , respectively. Here, antifungal activity of harmol was characterized in terms of pH dependency and conidial targets; also photodynamic effects of UVA irradiation on the antimicrobial action were evaluated. Harmol was able to inhibit the growth of both post-harvest fungal disease agents only in acidic conditions (pH 5), when it was found in its protonated form. Conidia treated with harmol exhibited membrane integrity loss, cell wall disruption, and cytoplasm disorganization. All these deleterious effects were more evident for B. cinerea in comparison to P. digitatum . When conidial suspensions were irradiated with UVA in the presence of harmol, antimicrobial activity against both pathogens was enhanced, compared to non-irradiated conditions. B. cinerea exhibited a high intracellular production of reactive oxygen species (ROS) when was incubated with harmol in irradiated and non-irradiated treatments. P. digitatum showed a significant increase in ROS accumulation only when treated with photoexcited harmol. The present work contributes to unravel the antifungal activity of harmol and its photoexcited counterpart against phytopathogenic conidia, focusing on ROS accumulation which could account for damage on different cellular targets.

  4. Selection of endophytic fungi from comfrey (Symphytum officinale L.) for in vitro biological control of the phytopathogen Sclerotinia sclerotiorum (Lib.).

    Science.gov (United States)

    Rocha, Rafaeli; da Luz, Daniela Eleutério; Engels, Cibelle; Pileggi, Sônia Alvim Veiga; de Souza Jaccoud Filho, David; Matiello, Rodrigo Rodrigues; Pileggi, Marcos

    2009-01-01

    Biological control consists of using one organism to attack another that may cause economic damage to crops. Integrated Pest Management (IPM) is a very common strategy. The white mold produced by Sclerotinia sclerotiorum (Lib.) causes considerable damage to bean crops. This fungus is a soil inhabitant, the symptoms of which are characterized by water-soaked lesions covered by a white cottony fungal growth on the soil surface and/or the host plant. Possible biological control agents taken from plants are being investigated as phytopathogen inhibitors. These are endophytic microorganisms that inhabit the intercellular spaces of vegetal tissues and are often responsible for antimicrobial production. The objective of the present study was to select endophytic fungi isolated from comfrey (Symphytum officinale L.) leaves with in vitro antagonist potential against the phytopathogenic fungus S. sclerotiorum. Twelve isolates of endophytic fungi and a pathogenic strain of S. sclerotiorum were used in the challenge method. With the aid of this method, four endophytes with the best antagonistic activity against S. sclerotiorum were selected. Pathogen growth inhibition zones were considered indicative of antibiosis. The percentages of pathogenic mycelia growth were measured both with and without the antagonist, resulting in growth reductions of 46.7% to 50.0% for S. sclerotiorum. These analyses were performed by evaluating the endophytic/pathogenic mycelia growth in mm/day over an eight-day period of antagonistic tests.

  5. Virulence of the Phytopathogen Pseudomonas syringae pv. Maculicola Is rpoN Dependent

    Science.gov (United States)

    Hendrickson, Erik L.; Guevera, Pablo; Peñaloza-Vàzquez, Alejandro; Shao, Jing; Bender, Carol; Ausubel, Frederick M.

    2000-01-01

    We cloned the rpoN (ntrA and glnF) gene encoding ς54 from the phytopathogen Pseudomonas syringae pv. maculicola strain ES4326. The P. syringae ES4326 rpoN gene complemented Pseudomonas aeruginosa, Escherichia coli, and Klebsiella aerogenes rpoN mutants for a variety of rpoN mutant phenotypes, including the inability to utilize nitrate as sole nitrogen source. DNA sequence analysis of the P. syringae ES4326 rpoN gene revealed that the deduced amino acid sequence was most similar (86% identity; 95% similarity) to the ς54 protein encoded by the Pseudomonas putida rpoN gene. A marker exchange protocol was used to construct an ES4326 rpoN insertional mutation, rpoN::Kmr. In contrast to wild-type ES4326, ES4326 rpoN::Kmr was nonmotile and could not utilize nitrate, urea, C4-dicarboxylic acids, several amino acids, or concentrations of ammonia below 2 mM as nitrogen sources. rpoN was essential for production of the phytotoxin coronatine and for expression of the structural genes encoding coronamic acid. In addition, ES4326 rpoN::Kmr did not multiply or elicit disease symptoms when infiltrated into Arabidopsis thaliana leaves, did not elicit the accumulation of several Arabidopsis defense-related mRNAs, and did not elicit a hypersensitive response (HR) when infiltrated into tobacco (Nicotiana tabacum) leaves. Furthermore, whereas P. syringae ES4326 carrying the avirulence gene avrRpt2 elicited an HR when infiltrated into Arabidopsis ecotype Columbia leaves, ES4326 rpoN::Kmr carrying avrRpt2 elicited no response. Constitutive expression of ES4326 hrpL in ES4326 rpoN::Kmr partially restored defense-related mRNA accumulation, showing a direct role for the hrp cluster in host defense gene induction in a compatible host-pathogen interaction. However, constitutive expression of hrpL in ES4326 rpoN::Kmr did not restore coronatine production, showing that coronatine biosynthesis requires factors other than hrpL. PMID:10852883

  6. Antifungal activity of essential oil isolated from Ocimum gratissimum L. (eugenol chemotype against phytopathogenic fungi

    Directory of Open Access Journals (Sweden)

    Terezinha de Jesus Faria

    2006-11-01

    Full Text Available An investigation of antifungal activity of the essential oil obtained by steam-distillation (1.1% w/w of the aerial parts of Ocimum gratissimum and of an ethanolic extract from the steam-distillation residue was carried out using the agar diffusion method. The results revealed that the essential oil inhibited the growth of all fungi tested, including the phytopathogens, Botryosphaeria rhodina, Rhizoctonia sp. and two strains of Alternaria sp., while the extract from the residue was inactive. The essential oil was subjected to TLC bioautography used to detect fungitoxic constituents. The compound that showed antifungal activity was isolated and identified as eugenol. GC/MS analysis showed that eugenol was the main constituent of the essential oil studied. The antifungal activity of eugenol was evaluated against a species of Alternaria isolated from tomato (A1 and Penicillium chrysogenum. The minimal inhibitory concentrations of eugenol were 0.16 and 0.31 mg/disc for Alternaria sp. (A1 and P. chrysogenum, respectively.O óleo essencial resultante da destilação por arraste a vapor das partes aéreas de Ocimum gratissimum e o extrato etanólico obtido do resíduo da destilação foram avaliados quanto à atividade antifúngica, utilizando-se o método de difusão em agar. O óleo essencial inibiu o crescimento de todos os fungos testados, incluindo os fitopatogênicos Botryosphaeria rhodina e duas espécies de Alternaria sp, enquanto que o extrato do resíduo da destilação não apresentou atividade. O óleo essencial foi, então, submetido ao método de bioautografia em TLC para detecção do composto ativo. O componente ativo foi isolado e identificado através da análise por cromatografia gasosa acoplada à espectrometria de massas como o eugenol, constituinte majoritário do óleo estudado. Ensaios de atividade antifúngica revelaram a atividade do eugenol contra Alternaria isolada de tomate (A1 e Penicillium chrysogenum. As concentra

  7. Pathogen and biological contamination management in plant tissue culture: phytopathogens, vitro pathogens, and vitro pests.

    Science.gov (United States)

    Cassells, Alan C

    2012-01-01

    The ability to establish and grow plant cell, organ, and tissue cultures has been widely exploited for basic and applied research, and for the commercial production of plants (micro-propagation). Regardless of whether the application is for research or commerce, it is essential that the cultures be established in vitro free of biological contamination and be maintained as aseptic cultures during manipulation, growth, and storage. The risks from microbial contamination are spurious experimental results due to the effects of latent contaminants or losses of valuable experimental or commercial cultures. Much of the emphasis in culture contamination management historically focussed on the elimination of phytopathogens and the maintenance of cultures free from laboratory contamination by environmental bacteria, fungi (collectively referred to as "vitro pathogens", i.e. pathogens or environmental micro-organisms which cause culture losses), and micro-arthropods ("vitro pests"). Microbial contamination of plant tissue cultures is due to the high nutrient availability in the almost universally used Murashige and Skoog (Physiol Plant 15:473-497, 1962) basal medium or variants of it. In recent years, it has been shown that many plants, especially perennials, are at least locally endophytically colonized intercellularly by bacteria. The latter, and intracellular pathogenic bacteria and viruses/viroids, may pass latently into culture and be spread horizontally and vertically in cultures. Growth of some potentially cultivable endophytes may be suppressed by the high salt and sugar content of the Murashige and Skoog basal medium and suboptimal temperatures for their growth in plant tissue growth rooms. The management of contamination in tissue culture involves three stages: disease screening (syn. disease indexing) of the stock plants with disease and endophyte elimination where detected; establishment and pathogen and contaminant screening of established initial cultures

  8. Identification and characterization of integron-mediated antibiotic resistance in the phytopathogen Xanthomonas oryzae pv. oryzae.

    Directory of Open Access Journals (Sweden)

    Ying Xu

    report of resistance integron in a phytopathogenic bacteria.

  9. Identification and Characterization of Integron-Mediated Antibiotic Resistance in the Phytopathogen Xanthomonas oryzae pv. oryzae

    Science.gov (United States)

    Zhou, Ming-guo

    2013-01-01

    integron in a phytopathogenic bacteria. PMID:23437082

  10. 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

  11. Detección de Erwinia chrysanthemi pv zeae (Sabet 1954) Victoria et al., 1975 en maíz en Morelos, México.

    OpenAIRE

    Durán Peralta, Elisa

    2011-01-01

    La “pudrición del tallo del maíz” causada por Erwinia chrysanthemi pv zeae es una de las principales enfermedades del maíz (Zea mays L.) en los países tropicales y subtropicales. En el presente trabajo se realizó la extracción de ADN del patógeno y se realizó PCR utilizando los iniciadores ADE1 y ADE2, diseñados para E. chrysanthemi (Burkh.) Young et al. 1978, que amplifican un fragmento de 420pb del gen pelADE. Se demostró que estos también pueden ser utilizados para su detección por PC...

  12. Identification des souches pathogènes d'Erwinia amylovora par inoculation de cals de racine de Malus domestica, cv. " Golden Delicious "

    OpenAIRE

    PAULIN, Jean-Pierre; DURON, Michel

    1986-01-01

    La bactérie Erwinia amylovora, agent du feu bactérien des Pomoïdeae, a été inoculée sur cals de racine de pommier (cv. "Golden Delicious "). Le résultat de cette inoculation est la production de gouttes d’exsudat caractéristiques de la maladie. Cette réponse, qui n’est obtenue qu’avec la forme virulente du pathogène, est spécifique : sous réserve de respecter certaines conditions expérimentales (en particulier taux d’inoculum inférieur à 104 germes/ml), aucune des 61 autres souches tes...

  13. Antimicrobial Activity of Bacillus sp. Natural Isolates and Their Potential Use in the Biocontrol of Phytopathogenic Bacteria

    Directory of Open Access Journals (Sweden)

    Tanja Berić

    2012-01-01

    Full Text Available Screening of 203 Bacillus sp. natural isolates for antimicrobial activity against phytopathogenic bacteria showed that 127 tested strains inhibit at least one sensitive strain, which illustrates their potential use as biocontrol agents. Among them, 104 isolates showed significant antagonism against Xanthomonas oryzae pv. oryzae, and only one of these (VPS50.2 synthesizes bacteriocin. An additional screening tested whether 51 isolates contained genes involved in the biosynthesis of lipopeptides of the iturin and surfactin classes. Results show that 33 isolates harbour the operon for iturin biosynthesis, and six of them carry the sfp gene, responsible for the biosynthesis of surfactin. Lipopeptide purification from the supernatant of isolate SS12.9 (identified as B. subtilis or B. amyloliquefaciens was performed using ethyl acetate extraction, ultrafiltration and reversed phase HPLC. Mass spectrometry analysis confirmed that isolate SS12.9 produces a substance of the iturin class with potential for biocontrol of X. oryzae pv. oryzae.

  14. 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

  15. Do induced responses mediate the ecological interactions between the specialist herbivores and phytopathogens of an alpine plant?

    Science.gov (United States)

    Röder, Gregory; Rahier, Martine; Naisbit, Russell E

    2011-05-04

    Plants are not passive victims of the myriad attackers that rely on them for nutrition. They have a suite of physical and chemical defences, and are even able to take advantage of the enemies of their enemies. These strategies are often only deployed upon attack, so may lead to indirect interactions between herbivores and phytopathogens. In this study we test for induced responses in wild populations of an alpine plant (Adenostyles alliariae) that possesses constitutive chemical defence (pyrrolizidine alkaloids) and specialist natural enemies (two species of leaf beetle, Oreina elongata and Oreina cacaliae, and the phytopathogenic rust Uromyces cacaliae). Plants were induced in the field using chemical elicitors of the jasmonic acid (JA) and salicylic acid (SA) pathways and monitored for one month under natural conditions. There was evidence for induced resistance, with lower probability and later incidence of attack by beetles in JA-induced plants and of rust infection in SA-induced plants. We also demonstrate ecological cross-effects, with reduced fungal attack following JA-induction, and a cost of SA-induction arising from increased beetle attack. As a result, there is the potential for negative indirect effects of the beetles on the rust, while in the field the positive indirect effect of the rust on the beetles appears to be over-ridden by direct effects on plant nutritional quality. Such interactions resulting from induced susceptibility and resistance must be considered if we are to exploit plant defences for crop protection using hormone elicitors or constitutive expression. More generally, the fact that induced defences are even found in species that possess constitutively-expressed chemical defence suggests that they may be ubiquitous in higher plants.

  16. Multiple interkingdom horizontal gene transfers in Pyrenophora and closely related species and their contributions to phytopathogenic lifestyles.

    Directory of Open Access Journals (Sweden)

    Bao-Fa Sun

    Full Text Available Many studies have reported horizontal gene transfer (HGT events from eukaryotes, especially fungi. However, only a few investigations summarized multiple interkingdom HGTs involving important phytopathogenic species of Pyrenophora and few have investigated the genetic contributions of HGTs to fungi. We investigated HGT events in P. teres and P. tritici-repentis and discovered that both species harbored 14 HGT genes derived from bacteria and plants, including 12 HGT genes that occurred in both species. One gene coding a leucine-rich repeat protein was present in both species of Pyrenophora and it may have been transferred from a host plant. The transfer of genes from a host plant to pathogenic fungi has been reported rarely and we discovered the first evidence for this transfer in phytopathogenic Pyrenophora. Two HGTs in Pyrenophora underwent subsequent duplications. Some HGT genes had homologs in a few other fungi, indicating relatively ancient transfer events. Functional analyses indicated that half of the HGT genes encoded extracellular proteins and these may have facilitated the infection of plants by Pyrenophora via interference with plant defense-response and the degradation of plant cell walls. Some other HGT genes appeared to participate in carbohydrate metabolism. Together, these functions implied that HGTs may have led to highly efficient mechanisms of infection as well as the utilization of host carbohydrates. Evolutionary analyses indicated that HGT genes experienced amelioration, purifying selection, and accelerated evolution. These appeared to constitute adaptations to the background genome of the recipient. The discovery of multiple interkingdom HGTs in Pyrenophora, their significance to infection, and their adaptive evolution, provided valuable insights into the evolutionary significance of interkingdom HGTs from multiple donors.

  17. Do induced responses mediate the ecological interactions between the specialist herbivores and phytopathogens of an alpine plant?

    Directory of Open Access Journals (Sweden)

    Gregory Röder

    2011-05-01

    Full Text Available Plants are not passive victims of the myriad attackers that rely on them for nutrition. They have a suite of physical and chemical defences, and are even able to take advantage of the enemies of their enemies. These strategies are often only deployed upon attack, so may lead to indirect interactions between herbivores and phytopathogens. In this study we test for induced responses in wild populations of an alpine plant (Adenostyles alliariae that possesses constitutive chemical defence (pyrrolizidine alkaloids and specialist natural enemies (two species of leaf beetle, Oreina elongata and Oreina cacaliae, and the phytopathogenic rust Uromyces cacaliae. Plants were induced in the field using chemical elicitors of the jasmonic acid (JA and salicylic acid (SA pathways and monitored for one month under natural conditions. There was evidence for induced resistance, with lower probability and later incidence of attack by beetles in JA-induced plants and of rust infection in SA-induced plants. We also demonstrate ecological cross-effects, with reduced fungal attack following JA-induction, and a cost of SA-induction arising from increased beetle attack. As a result, there is the potential for negative indirect effects of the beetles on the rust, while in the field the positive indirect effect of the rust on the beetles appears to be over-ridden by direct effects on plant nutritional quality. Such interactions resulting from induced susceptibility and resistance must be considered if we are to exploit plant defences for crop protection using hormone elicitors or constitutive expression. More generally, the fact that induced defences are even found in species that possess constitutively-expressed chemical defence suggests that they may be ubiquitous in higher plants.

  18. Draft Genome Sequence of Bacillus velezensis 3A-25B, a Strain with Biocontrol Activity against Fungal and Oomycete Root Plant Phytopathogens, Isolated from Grassland Soil.

    Science.gov (United States)

    Martínez-Raudales, Inés; De La Cruz-Rodríguez, Yumiko; Vega-Arreguín, Julio; Alvarado-Gutiérrez, Alejandro; Fraire-Mayorga, Atzin; Alvarado-Rodríguez, Miguel; Balderas-Hernández, Victor; Gómez-Soto, José Manuel; Fraire-Velázquez, Saúl

    2017-09-28

    Here, we present the draft genome of Bacillus velezensis 3A-25B, which totaled 4.01 Mb with 36 contigs, 3,948 genes, and a GC content of 46.34%. This strain, which demonstrates biocontrol activity against root rot causal phytopathogens in horticultural crops and friendly interactions in roots of pepper plantlets, was obtained from grassland soil in Zacatecas Province, Mexico. Copyright © 2017 Martínez-Raudales et al.

  19. Green synthesis of protein capped silver nanoparticles from phytopathogenic fungus Macrophomina phaseolina (Tassi) Goid with antimicrobial properties against multidrug-resistant bacteria

    OpenAIRE

    Chowdhury, Supriyo; Basu, Arpita; Kundu, Surekha

    2014-01-01

    In recent years, green synthesis of nanoparticles, i.e., synthesizing nanoparticles using biological sources like bacteria, algae, fungus, or plant extracts have attracted much attention due to its environment-friendly and economic aspects. The present study demonstrates an eco-friendly and low-cost method of biosynthesis of silver nanoparticles using cell-free filtrate of phytopathogenic fungus Macrophomina phaseolina. UV-visible spectrum showed a peak at 450 nm corresponding to the plasmon ...

  20. Expression of antimicrobial peptides thanatin(S) in transgenic Arabidopsis enhanced resistance to phytopathogenic fungi and bacteria.

    Science.gov (United States)

    Wu, Tingquan; Tang, Dingzhong; Chen, Weida; Huang, Hexun; Wang, Rui; Chen, Yongfang

    2013-09-15

    Thanatin(S) is an analog of thanatin, an insect antimicrobial peptide possessing strong and broad spectrum of antimicrobial activity. In order to investigate if the thanatin could be used in engineering transgenic plants for increased resistance against phytopathogens, the synthetic thanatin(S) was introduced into Arabidopsis thaliana plants. To increase the expression level of thanatin(S) in plants, the coding sequence was optimized by plant-preference codon. To avoid cellular protease degradation, signal peptide of rice Cht1 was fused to N terminal of thanatin(S) for secreting the expressed thanatin(S) into intercellular spaces. To evaluate the application value of thanatin(S) in plant disease control, the synthesized coding sequence of Cht1 signal peptide (Cht1SP)-thanatin(S) was ligated to plant gateway destination binary vectors pGWB11 (with FLAG tag). Meanwhile, in order to observe the subcellular localization of Cht1SP-thanatin(S)-GFP and thanatin(S)-GFP, the sequences of Cht1SP-thanatin(S) and thanatin(S) were respectively linked to pGWB5 (with GFP tag). The constructs were transformed into Arabidopsis ecotype Col-0 and mutant pad4-1 via Agrobacterium-mediated transformation. The transformants with Cht1SP-thanatin(S)-FLAG fusion gene were analyzed by genomic PCR, real-time PCR, and western blots and the transgenic Arabidopsis plants introduced respectively Cht1SP-thanatin(S)-GFP and thanatin(S)-GFP were observed by confocal microscopy. Transgenic plants expressing Cht1SP-thanatin(S)-FLAG fusion protein showed antifungal activity against Botrytis cinerea and powdery mildew, as well as antibacterial activity against Pseudomonas syringae pv. tomato. And the results from confocal observation showed that the GFP signal from Cht1SP-thanatin(S)-GFP transgenic Arabidopsis plants occurred mainly in intercellular space, while that from thanatin(S)-GFP transgenic plants was mainly detected in the cytoplasm and that from empty vector transgenic plants was distributed

  1. Marine Actinobacteria as a source of compounds for phytopathogen control: An integrative metabolic-profiling / bioactivity and taxonomical approach.

    Directory of Open Access Journals (Sweden)

    Luz A Betancur

    Full Text Available Marine bacteria are considered as promising sources for the discovery of novel biologically active compounds. In this study, samples of sediment, invertebrate and algae were collected from the Providencia and Santa Catalina coral reef (Colombian Caribbean Sea with the aim of isolating Actinobateria-like strain able to produce antimicrobial and quorum quenching compounds against pathogens. Several approaches were used to select actinobacterial isolates, obtaining 203 strains from all samples. According to their 16S rRNA gene sequencing, a total of 24 strains was classified within Actinobacteria represented by three genera: Streptomyces, Micromonospora, and Gordonia. In order to assess their metabolic profiles, the actinobacterial strains were grown in liquid cultures, and LC-MS-based analyses from ethyl acetate fractions were performed. Based on taxonomical classification, screening information of activity against phytopathogenic strains and quorum quenching activity, as well as metabolic profiling, six out of the 24 isolates were selected for follow-up with chemical isolation and structure identification analyses of putative metabolites involved in antimicrobial activities.

  2. Synthesis and biological evaluation of novel fluorine-containing stilbene derivatives as fungicidal agents against phytopathogenic fungi.

    Science.gov (United States)

    Jian, Weilin; He, Daohang; Xi, Pinggen; Li, Xinwei

    2015-11-18

    The rising development of resistance to conventional fungicides is driving the search for new alternative candidates to control plant diseases. In this study, a series of new fluorine-containing stilbene derivatives was synthesized on the basis of our previous quantitative structure-activity relationship analysis results. Bioassays in vivo revealed that the title compounds exhibited potent fungicidal activities against phytopathogenic fungi (Colletotrichum lagenarium and Pseudoperonospora cubensis) from cucumber plants. In comparison to the previous results, the introduction of a fluorine moiety showed improved activities of some compounds against those fungi. Notably, compound 9 exhibited a control efficacy against C. lagenarium (83.4 ± 1.3%) comparable to that of commercial fungicide (82.7 ± 1.7%). For further understanding the possible mode of action of the stilbene against C. lagenarium, the effects on hyphal morphology, electrolyte leakage, and respiration of mycelial cell suspension were studied. Microscopic observation showed considerably deformed mycelial morphology. The conductivity of mycelial suspension increased in the presence of compound 9, whereas no significantly inhibitory effect on respiration was observed. Taken together, the fungicidal mechanism of this stilbene is associated with its membrane disruption effect, resulting in increased membrane permeability. These results provide important clues for mechanistic study and derivatization of stilbenes as alternative sources of fungicidal agents for plant disease control.

  3. Marine Actinobacteria as a source of compounds for phytopathogen control: An integrative metabolic-profiling / bioactivity and taxonomical approach

    Science.gov (United States)

    Betancur, Luz A.; Naranjo-Gaybor, Sandra J.; Vinchira-Villarraga, Diana M.; Moreno-Sarmiento, Nubia C.; Maldonado, Luis A.; Suarez-Moreno, Zulma R.; Acosta-González, Alejandro; Padilla-Gonzalez, Gillermo F.; Puyana, Mónica; Castellanos, Leonardo; Ramos, Freddy A.

    2017-01-01

    Marine bacteria are considered as promising sources for the discovery of novel biologically active compounds. In this study, samples of sediment, invertebrate and algae were collected from the Providencia and Santa Catalina coral reef (Colombian Caribbean Sea) with the aim of isolating Actinobateria-like strain able to produce antimicrobial and quorum quenching compounds against pathogens. Several approaches were used to select actinobacterial isolates, obtaining 203 strains from all samples. According to their 16S rRNA gene sequencing, a total of 24 strains was classified within Actinobacteria represented by three genera: Streptomyces, Micromonospora, and Gordonia. In order to assess their metabolic profiles, the actinobacterial strains were grown in liquid cultures, and LC-MS-based analyses from ethyl acetate fractions were performed. Based on taxonomical classification, screening information of activity against phytopathogenic strains and quorum quenching activity, as well as metabolic profiling, six out of the 24 isolates were selected for follow-up with chemical isolation and structure identification analyses of putative metabolites involved in antimicrobial activities. PMID:28225766

  4. Chitinase genes revealed and compared in bacterial isolates, DNA extracts and a metagenomic library from a phytopathogen suppressive soil

    Energy Technology Data Exchange (ETDEWEB)

    Hjort, K.; Bergstrom, M.; Adesina, M.F.; Jansson, J.K.; Smalla, K.; Sjoling, S.

    2009-09-01

    Soil that is suppressive to disease caused by fungal pathogens is an interesting source to target for novel chitinases that might be contributing towards disease suppression. In this study we screened for chitinase genes, in a phytopathogen-suppressive soil in three ways: (1) from a metagenomic library constructed from microbial cells extracted from soil, (2) from directly extracted DNA and (3) from bacterial isolates with antifungal and chitinase activities. Terminal-restriction fragment length polymorphism (T-RFLP) of chitinase genes revealed differences in amplified chitinase genes from the metagenomic library and the directly extracted DNA, but approximately 40% of the identified chitinase terminal-restriction fragments (TRFs) were found in both sources. All of the chitinase TRFs from the isolates were matched to TRFs in the directly extracted DNA and the metagenomic library. The most abundant chitinase TRF in the soil DNA and the metagenomic library corresponded to the TRF{sup 103} of the isolate, Streptomyces mutomycini and/or Streptomyces clavifer. There were good matches between T-RFLP profiles of chitinase gene fragments obtained from different sources of DNA. However, there were also differences in both the chitinase and the 16S rRNA gene T-RFLP patterns depending on the source of DNA, emphasizing the lack of complete coverage of the gene diversity by any of the approaches used.

  5. Microbial synthesis of chitinase in solid cultures and its potential as a biocontrol agent against phytopathogenic fungus Colletotrichum gloeosporioides.

    Science.gov (United States)

    Sandhya, Chandran; Binod, Parameswaran; Nampoothiri, K Madhavan; Szakacs, George; Pandey, Ashok

    2005-10-01

    Antifungal activity of chitinase can be effectively utilized in biologic pest control strategies. Because solid-state cultivation has been termed a cost-effective means for fungal growth and metabolite production, chitinase production by Trichoderma harzianum was studied using wheat bran-based solid medium containing 1% colloidal chitin. Chitinase synthesis was found to be growth associated because maximum enzyme (5.4 U/g of dry substrate) and biomass production occurred at 72 h. Substrate moisture had a critical impact on chitinase production; five grams of medium having an initial moisture content of 68.4% when incubated for 72 h increased the enzyme yield to 9.3 U/g of dry substrate. Optimization of colloidal chitin concentration showed that improvements in chitinase yield and maximum activity were attained with a 2% (w/w) concentration. Supplementation of additional nitrogen sources also influenced enzyme production, and the best yield was obtained with yeast extract. The effect of crude chitinase on hyphal morphology of the phytopathogenic fungus Colletotrichum gloeosporioides was swelling as well as lysis of hyphal wall, depending on the age of the mycelium. Studies of pH and thermal stability showed that crude culture filtrate was active over pH 4.0-6.0 and retained about 48.2% activity after 40 min of incubation at 40 degrees C.

  6. Antimicrobial activity of broccoli (Brassica oleracea var. italica) cultivar Avenger against pathogenic bacteria, phytopathogenic filamentous fungi and yeast.

    Science.gov (United States)

    Pacheco-Cano, R D; Salcedo-Hernández, R; López-Meza, J E; Bideshi, D K; Barboza-Corona, J E

    2018-01-01

    The objective of this study was to show whether the edible part of broccoli has antibacterial and antifungal activity against micro-organism of importance in human health and vegetable spoilage, and to test if this effect was partially due to antimicrobial peptides (AMPs). Crude extracts were obtained from florets and stems of broccoli cultivar Avenger and the inhibitory effect was demonstrated against pathogenic bacteria (Bacillus cereus, Staphylococcus xylosus, Staphylococcus aureus, Shigella flexneri, Shigella sonnei, Proteus vulgaris), phytopathogenic fungi (Colletotrichum gloeosporioides, Asperigillus niger) and yeasts (Candida albicans and Rhodotorula sp.). It was shown that samples treated with proteolytic enzymes had a reduction of approximately 60% in antibacterial activity against Staph. xylosus, suggesting that proteinaceous compounds might play a role in the inhibitory effect. Antimicrobial components in crude extracts were thermoresistant and the highest activity was observed under acidic conditions. It was shown that antifungal activity of broccoli's crude extracts might not be attributed to chitinases. Organic broccoli cultivar Avenger has antimicrobial activity against pathogenic bacteria, yeast and phytophatogenic fungi. Data suggest that this effect is partially due to AMPs. Broccoli's crude extracts have activity not only against pathogenic bacteria but also against phytophatogenic fungi of importance in agriculture. We suggest for first time that the inhibitory effect is probably due to AMPs. © 2017 The Society for Applied Microbiology.

  7. The Comprehensive Phytopathogen Genomics Resource: a web-based resource for data-mining plant pathogen genomes.

    Science.gov (United States)

    Hamilton, John P; Neeno-Eckwall, Eric C; Adhikari, Bishwo N; Perna, Nicole T; Tisserat, Ned; Leach, Jan E; Lévesque, C André; Buell, C Robin

    2011-01-01

    The Comprehensive Phytopathogen Genomics Resource (CPGR) provides a web-based portal for plant pathologists and diagnosticians to view the genome and trancriptome sequence status of 806 bacterial, fungal, oomycete, nematode, viral and viroid plant pathogens. Tools are available to search and analyze annotated genome sequences of 74 bacterial, fungal and oomycete pathogens. Oomycete and fungal genomes are obtained directly from GenBank, whereas bacterial genome sequences are downloaded from the A Systematic Annotation Package (ASAP) database that provides curation of genomes using comparative approaches. Curated lists of bacterial genes relevant to pathogenicity and avirulence are also provided. The Plant Pathogen Transcript Assemblies Database provides annotated assemblies of the transcribed regions of 82 eukaryotic genomes from publicly available single pass Expressed Sequence Tags. Data-mining tools are provided along with tools to create candidate diagnostic markers, an emerging use for genomic sequence data in plant pathology. The Plant Pathogen Ribosomal DNA (rDNA) database is a resource for pathogens that lack genome or transcriptome data sets and contains 131 755 rDNA sequences from GenBank for 17 613 species identified as plant pathogens and related genera. Database URL: http://cpgr.plantbiology.msu.edu.

  8. Complete sequence of a double-stranded RNA from the phytopathogenic fungus Erysiphe cichoracearum that might represent a novel endornavirus.

    Science.gov (United States)

    Du, Zhenguo; Lin, Wenzhong; Qiu, Ping; Liu, Xiaojuan; Guo, Lingfang; Wu, Kangcheng; Zhang, Songbai; Wu, Zujian

    2016-08-01

    A double-stranded RNA (dsRNA) HBJZ1506 recovered from the phytopathogenic fungus Erysiphe cichoracearum infecting Calendula officinalis in Jingzhou, Hubei Province, China, was sequenced. HBJZ1506 comprises 11,908 nucleotides (nt) and contains a 11,859-nt-long open reading frame (ORF) coding for a polypeptide that is 61 % identical to that of a putative endornavirus named grapevine endophyte endornavirus (GeEV). The putative polyprotein has an RNA-dependent RNA polymerase (RdRp) domain and an RNA helicase domain, which show homology to and have an arrangement that is similar to that of their counterparts in approved or putative endornaviruses. In a phylogenetic tree constructed using amino acid sequences of the RdRp region of HBJZ1506 and selected endornaviruses, HBJZ1506 clustered with endornaviruses and formed a well-supported monophyletic branch with GeEV. These results suggest that HBJZ1506 might represent a novel endornavirus, for which the name Erysiphe cichoracearum endornavirus (EcEV) is proposed.

  9. Genome, Proteome and Structure of a T7-Like Bacteriophage of the Kiwifruit Canker Phytopathogen Pseudomonas syringae pv. actinidiae.

    Science.gov (United States)

    Frampton, Rebekah A; Acedo, Elena Lopez; Young, Vivienne L; Chen, Danni; Tong, Brian; Taylor, Corinda; Easingwood, Richard A; Pitman, Andrew R; Kleffmann, Torsten; Bostina, Mihnea; Fineran, Peter C

    2015-06-24

    Pseudomonas syringae pv. actinidiae is an economically significant pathogen responsible for severe bacterial canker of kiwifruit (Actinidia sp.). Bacteriophages infecting this phytopathogen have potential as biocontrol agents as part of an integrated approach to the management of bacterial canker, and for use as molecular tools to study this bacterium. A variety of bacteriophages were previously isolated that infect P. syringae pv. actinidiae, and their basic properties were characterized to provide a framework for formulation of these phages as biocontrol agents. Here, we have examined in more detail φPsa17, a phage with the capacity to infect a broad range of P. syringae pv. actinidiae strains and the only member of the Podoviridae in this collection. Particle morphology was visualized using cryo-electron microscopy, the genome was sequenced, and its structural proteins were analysed using shotgun proteomics. These studies demonstrated that φPsa17 has a 40,525 bp genome, is a member of the T7likevirus genus and is closely related to the pseudomonad phages φPSA2 and gh-1. Eleven structural proteins (one scaffolding) were detected by proteomics and φPsa17 has a capsid of approximately 60 nm in diameter. No genes indicative of a lysogenic lifecycle were identified, suggesting the phage is obligately lytic. These features indicate that φPsa17 may be suitable for formulation as a biocontrol agent of P. syringae pv. actinidiae.

  10. Transgenic Brassica juncea plants expressing MsrA1, a synthetic cationic antimicrobial peptide, exhibit resistance to fungal phytopathogens.

    Science.gov (United States)

    Rustagi, Anjana; Kumar, Deepak; Shekhar, Shashi; Yusuf, Mohd Aslam; Misra, Santosh; Sarin, Neera Bhalla

    2014-06-01

    Cationic antimicrobial peptides (CAPs) have shown potential against broad spectrum of phytopathogens. Synthetic versions with desirable properties have been modeled on these natural peptides. MsrA1 is a synthetic chimera of cecropin A and melittin CAPs with antimicrobial properties. We generated transgenic Brassica juncea plants expressing the msrA1 gene aimed at conferring fungal resistance. Five independent transgenic lines were evaluated for resistance to Alternaria brassicae and Sclerotinia sclerotiorum, two of the most devastating pathogens of B. juncea crops. In vitro assays showed inhibition by MsrA1 of Alternaria hyphae growth by 44-62 %. As assessed by the number and size of lesions and time taken for complete leaf necrosis, the Alternaria infection was delayed and restricted in the transgenic plants with the protection varying from 69 to 85 % in different transgenic lines. In case of S. sclerotiorum infection, the lesions were more severe and spread profusely in untransformed control compared with transgenic plants. The sclerotia formed in the stem of untransformed control plants were significantly more in number and larger in size than those present in the transgenic plants where disease protection of 56-71.5 % was obtained. We discuss the potential of engineering broad spectrum biotic stress tolerance by transgenic expression of CAPs in crop plants.

  11. Genome, Proteome and Structure of a T7-Like Bacteriophage of the Kiwifruit Canker Phytopathogen Pseudomonas Syringae pv. Actinidiae

    Directory of Open Access Journals (Sweden)

    Rebekah A. Frampton

    2015-06-01

    Full Text Available Pseudomonas syringae pv. actinidiae is an economically significant pathogen responsible for severe bacterial canker of kiwifruit (Actinidia sp.. Bacteriophages infecting this phytopathogen have potential as biocontrol agents as part of an integrated approach to the management of bacterial canker, and for use as molecular tools to study this bacterium. A variety of bacteriophages were previously isolated that infect P. syringae pv. actinidiae, and their basic properties were characterized to provide a framework for formulation of these phages as biocontrol agents. Here, we have examined in more detail φPsa17, a phage with the capacity to infect a broad range of P. syringae pv. actinidiae strains and the only member of the Podoviridae in this collection. Particle morphology was visualized using cryo-electron microscopy, the genome was sequenced, and its structural proteins were analysed using shotgun proteomics. These studies demonstrated that φPsa17 has a 40,525 bp genome, is a member of the T7likevirus genus and is closely related to the pseudomonad phages φPSA2 and gh-1. Eleven structural proteins (one scaffolding were detected by proteomics and φPsa17 has a capsid of approximately 60 nm in diameter. No genes indicative of a lysogenic lifecycle were identified, suggesting the phage is obligately lytic. These features indicate that φPsa17 may be suitable for formulation as a biocontrol agent of P. syringae pv. actinidiae.

  12. Complete genome sequence of Bacillus amyloliquefaciens subsp. plantarum S499, a rhizobacterium that triggers plant defences and inhibits fungal phytopathogens.

    Science.gov (United States)

    Molinatto, Giulia; Puopolo, Gerardo; Sonego, Paolo; Moretto, Marco; Engelen, Kristof; Viti, Carlo; Ongena, Marc; Pertot, Ilaria

    2016-11-20

    Bacillus amyloliquefaciens subsp. plantarum S499 is a plant beneficial rhizobacterium with a good antagonistic potential against phytopathogens through the release of active secondary metabolites. Moreover, it can induce systemic resistance in plants by producing considerable amounts of surfactins. The complete genome sequence of B. amyloliquefaciens subsp. plantarum S499 includes a circular chromosome of 3,927,922bp and a plasmid of 8,008bp. A remarkable abundance in genomic regions of putative horizontal origin emerged from the analysis. Furthermore, we highlighted the presence of genes involved in the establishment of interactions with the host plants at the root level and in the competition with other soil-borne microorganisms. More specifically, genes related to the synthesis of amylolysin, amylocyclicin, and butirosin were identified. These antimicrobials were not known before to be part of the antibiotic arsenal of the strain. The information embedded in the genome will support the upcoming studies regarding the application of B. amyloliquefaciens isolates as plant-growth promoters and biocontrol agents. Copyright © 2016 Elsevier B.V. All rights reserved.

  13. Crystallographic structure and substrate-binding interactions of the molybdate-binding protein of the phytopathogen Xanthomonas axonopodis pv. citri.

    Science.gov (United States)

    Balan, Andrea; Santacruz-Pérez, Carolina; Moutran, Alexandre; Ferreira, Luís Carlos Souza; Neshich, Goran; Gonçalves Barbosa, João Alexandre Ribeiro

    2008-02-01

    In Xanthomonas axonopodis pv. citri (Xac or X. citri), the modA gene codes for a periplasmic protein (ModA) that is capable of binding molybdate and tungstate as part of the ABC-type transporter required for the uptake of micronutrients. In this study, we report the crystallographic structure of the Xac ModA protein with bound molybdate. The Xac ModA structure is similar to orthologs with known three-dimensional structures and consists of two nearly symmetrical domains separated by a hinge region where the oxyanion-binding site lies. Phylogenetic analysis of different ModA orthologs based on sequence alignments revealed three groups of molybdate-binding proteins: bacterial phytopathogens, enterobacteria and soil bacteria. Even though the ModA orthologs are segregated into different groups, the ligand-binding hydrogen bonds are mostly conserved, except for Archaeglobus fulgidus ModA. A detailed discussion of hydrophobic interactions in the active site is presented and two new residues, Ala38 and Ser151, are shown to be part of the ligand-binding pocket.

  14. 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

  15. Drosophila larvae food intake cessation following exposure to Erwinia contaminated media requires odor perception, Trpa1 channel and evf virulence factor.

    Science.gov (United States)

    Keita, Seydou; Masuzzo, Ambra; Royet, Julien; Kurz, C Leopold

    2017-05-01

    When exposed to microorganisms, animals use several protective strategies. On one hand, as elegantly exemplified in Drosophila melanogaster, the innate immune system recognizes microbial compounds and triggers an antimicrobial response. On the other hand, behaviors preventing an extensive contact with the microbes and thus reducing the risk of infection have been described. However, these reactions ranging from microbes aversion to intestinal transit increase or food intake decrease have been rarely defined at the molecular level. In this study, we set up an experimental system that allowed us to rapidly identify and quantify food intake decreases in Drosophila larvae exposed to media contaminated with bacteria. Specifically, we report a robust dose-dependent food intake decrease following exposure to the bacteria Erwinia carotovora carotovora strain Ecc15. We demonstrate that this response does not require Imd innate immune pathway, but rather the olfactory neuronal circuitry, the Trpa1 receptor and the evf virulence factor. Finally, we show that Ecc15 induce the same behavior in the invasive pest insect Drosophila suzukii. Copyright © 2017 Elsevier Ltd. All rights reserved.

  16. 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

    2013-01-01

    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. PMID:23770912

  17. 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.

  18. 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, D.A.

    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.

  19. Separation of Escherichia adecarboxylata from the "Erwinia herbicola-Enterobacter agglomerans" complex and from the other Enterobacteriaceae by nucleic acid and protein electrophoretic techniques.

    Science.gov (United States)

    Izard, D; Mergaert, J; Gavini, F; Beji, A; Kersters, K; De Ley, J; Leclerc, H

    1985-01-01

    The species Escherichia adecarboxylata was examined for DNA relatedness to the "Erwinia herbicola-Enterobacter agglomerans" complex and to other members of the family Enterobacteriaceae. DNA-DNA hybridizations (nitrocellulose filter method) showed that strains received as E. adecarboxylata were highly related to each other (73-100% homology). Three strains of E. agglomerans and one strain of E. herbicola showed, respectively, 77, 96, 97 and 92% relatedness with the labelled DNA of E. adecarboxylata. Two groups (E2 and E3) of "atypical coliforms" previously described by Gavini et al. (1983) showed high reassociation values (76-79% and 80-89%, respectively) with E. adecarboxylata. Most of these strains produced similar or nearly identical protein electrophoregrams. All these strains were therefore classified in E. adecarboxylata. This taxon yielded hybridization values lower than 53% with the previously described phenetic or genetic groups belonging to or related to the "herbicola-agglomerans" complex and values lower than 64% with 56 other species of the Enterobacteriaceae. It was concluded that E. adecarboxylata is a species different from E. agglomerans and the other species of the family Enterobacteriaceae. A new definition of the species E. adecarboxylata is presented.

  20. A putative three-dimensional targeting motif of polygalacturonase (PehA), a protein secreted through the type II (GSP) pathway in Erwinia carotovora.

    Science.gov (United States)

    Palomäki, Tiina; Pickersgill, Richard; Riekki, Ruusu; Romantschuk, Martin; Saarilahti, Hannu T

    2002-02-01

    Intramolecular information specifying protein secretion through the type II (GSP) pathway of Gram-negative bacteria was investigated. Two regions of the polygalacturonase (PehA) of Erwinia carotovora containing residues proposed to be included in a targeting motif were located, one close to the C-terminus between residues 342 and 369 and another between residues 84 and 135 in the large central loops. The regions were required together to promote secretion. Further residues in the middle of the protein were required for proper positioning of the regions, suggesting that they were both involved in interaction with the GSP. To our knowledge, this is the first time that a possible three-dimensional targeting motif has been defined. At least one of the motifs comprises a cluster on the surface of the protein. The two motifs are structurally dissimilar, suggesting that there are two distinct recognition regions in the GSP apparatus. Finally, we propose that the targeting motifs are of a complex conformational nature with some variability accommodated, as illustrated by the observation that many mutations exhibited no clear phenotype individually but, in combination, severely compromised secretion.

  1. 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. Copyright © 2012 Elsevier Ireland Ltd. All rights reserved.

  2. Evidence for growth of strains of the plant epiphytic bacterium Erwinia herbicola and transconjugation among the bacterial strains in guts of the silkworm Bombyx mori.

    Science.gov (United States)

    Watanabe, K; Hara, W; Sato, M

    1998-09-01

    Growth of plant epiphytic bacteria Erwinia herbicola and Pseudomonas syringae in guts of the silkworm, Bombyx mori, was studied. Fifth instar silkworm larvae were fed artificial diets supplemented with these bacteria for 6 to 12 h followed by uncontaminated diets. At 1, 3, and 6 days after feeding, bacteria were isolated from insect guts and feces. A much larger population of E. herbicola was detected in the samples collected 3 and 6 days after the inoculation than in samples collected after 1 day, indicating that these bacteria grew in the insect gut, while P. syringae was unable to survive. Transconjugation between E. herbicola strains in the insect gut was also examined. First, either a donor or a recipient strain was fed to the insects in artificial diets containing the bacteria during 12 h, and then pairing strains were fed during 12 h after starvation for 12 h. The conjugative plasmid pBPW1::Tn7 was transferred into recipient cells at very high frequencies (10(-1)/recipient after 3 days and 10(-3) after 6 days) in insect guts. Indigenous plasmids of E. herbicola mobilized RSF1010 plasmid into recipient cells at frequencies of 10(-4) in insect guts. These transconjugants were detected in the feces of the insects. Thus, plasmid-mediated gene transfer among the epiphytic bacteria in insect guts was demonstrated. The results obtained suggest that in insecta gene transfer may play an important role in the evolution of plant epiphytic bacteria. Copyright 1998 Academic Press.

  3. Plasmid-mediated gene transfer between insect-resident bacteria, Enterobacter cloacae, and plant-epiphytic bacteria, Erwinia herbicola, in guts of silkworm larvae.

    Science.gov (United States)

    Watanabe, K; Sato, M

    1998-11-01

    Five strains of Enterobacter cloacae isolated from several species of plants and insects were able to grow in the guts of silkworm larvae. A much larger population of Ent. cloacae strains was detected in the insect guts and feces collected 3 and 6 days than in samples collected 1 day after feeding artificial diets contaminating these bacteria. Furthermore, insect-origin strains of Ent. cloacae were mated with a donor strain, epiphytic Erwinia herbicola, harboring RSF1010 and pBPW1::Tn7 plasmids in the insect guts by introducing these bacteria through separate artificial diets administered at different times. A number of transconjugants, Ent. cloacae strains which had acquired RSF1010 plasmid, were detected from guts and fecal samples at transfer frequencies of 10(-2) to 10(-3) per recipient. Thus, gene transfer between epiphytic Er. herbicola and insect-resident Ent. cloacae strains in the insect guts was confirmed. These findings may provide significant information about the role of "in insecta mating" in the evolution of these bacteria.

  4. Characterization of Monoclonal Antibodies Specific for Erwinia carotovora subsp. atroseptica and Comparison of Serological Methods for Its Sensitive Detection on Potato Tubers

    Science.gov (United States)

    Gorris, María Teresa; Alarcon, Benito; Lopez, María M.; Cambra, Mariano

    1994-01-01

    Seven monoclonal antibodies (MAbs) to Erwinia carotovora subsp. atroseptica have been produced. One, called 4G4, reacted with high specificity for serogroup I of E. carotovora subsp. atroseptica, the most common serogroup on potato tubers in different serological assays. Eighty-six strains belonging to different E. carotovora subsp. atroseptica serogroups were assayed. Some strains of serogroup XXII also reacted positively. No cross-reactions were observed against other species of plant pathogenic bacteria or 162 saprophytic bacteria from potato tubers. Only one strain of E. chrysanthemi from potato cross-reacted. A comparison of several serological techniques to detect E. carotovora subsp. atroseptica on potato tubers was performed with MAb 4G4 or polyclonal antibodies. The organism was extracted directly from potato peels of artificially inoculated tubers by soaking or selective enrichment under anaerobiosis in a medium with polypectate. MAb 4G4 was able to detect specifically 240 E. carotovora subsp. atroseptica cells per ml by indirect immunofluorescence and immunofluorescence colony staining and after soaking by ELISA-DAS (double-antibody sandwich enzyme-linked immunosorbent assay) after enrichment. The same amount of cells was detected by using immunolectrotransfer with polyclonal antibodies, and E. carotovora subsp. atroseptica and subsp. carotovora were distinguished by the latter technique. ELISA-DAS using MAb 4G4 with an enrichment step also efficiently detected E. carotovora subsp. atroseptica in naturally infected tubers and plants. PMID:16349293

  5. Mutations in rpsL that confer streptomycin resistance show pleiotropic effects on virulence and the production of a carbapenem antibiotic in Erwinia carotovora.

    Science.gov (United States)

    Barnard, Anne M L; Simpson, Natalie J L; Lilley, Kathryn S; Salmond, George P C

    2010-04-01

    Spontaneous streptomycin-resistant derivatives of Erwinia carotovora subsp. carotovora strain ATTn10 were isolated. Sequencing of the rpsL locus (encoding the ribosomal protein S12) showed that each mutant was missense, with a single base change, resulting in the substitution of the wild-type lysine by arginine, threonine or asparagine at codon 43. Phenotypic analyses showed that the rpsL mutants could be segregated into two groups: K43R mutants showed reduced production of the beta-lactam secondary metabolite 1-carbapen-2-em-3 carboxylic acid (Car), but little effect on exoenzyme production or virulence in potato tuber tests. By contrast, the K43N and K43T mutations were pleiotropic, resulting in reduced exoenzyme production and virulence, as well as diminished Car production. The effect on Car production was due to reduced transcription of the quorum-sensing-dependent car biosynthetic genes. The effects of K43N and K43T mutations on Car production were partially alleviated by provision of an excess of the quorum-sensing signalling molecule N-(3-oxohexanoyl)-L-homoserine lactone. Finally, a proteomic analysis of the K43T mutant indicated that the abundance of a subset of intracellular proteins was affected by this rpsL mutation.

  6. Preservação de fungos fitopatogênicos habitantes do solo Storage of soilborne phytopathogenic fungi

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    César J. Bueno

    2006-03-01

    Full Text Available A preservação de fungos fitopatogênicos por longos períodos de tempo é importante para que pesquisas possam ser realizadas a qualquer momento. Os fungos habitantes do solo são organismos que podem produzir estruturas de resistência em face de situações adversas, tais como ausência de hospedeiros e ou condições climáticas desfavoráveis para a sua sobrevivência. O objetivo deste trabalho foi desenvolver metodologias de preservação de estruturas de resistência para os fungos Fusarium oxysporum f.sp. lycopersici raça 2, Macrophomina phaseolina, Rhizoctonia solani AG4 HGI, Sclerotium rolfsii, Sclerotinia sclerotiorum e Verticillium dahliae. O delineamento foi inteiramente casualizado, com um método de produção de estruturas para cada fungo, submetido a três tratamentos [temperatura ambiente de laboratório (28±2ºC, de geladeira (5ºC e de freezer (-20ºC] e com dois frascos por temperatura. Mensalmente, e por um período de um ano, a sobrevivência e o vigor das colônias de cada patógeno foram avaliadas em meios de cultura específicos. Testes de patogenicidade foram realizados após um ano de preservação, com as estruturas que sobreviveram aos melhores tratamentos (temperatura para todos os fungos. As melhores temperaturas (tratamentos para preservar os fungos foram: a F. oxysporum f.sp. lycopersici em temperatura de refrigeração e de freezer (5,2 e 2,9 x 10³ufc.g-1 de talco, respectivamente; b M. phaseolina em temperatura de refrigeração [100% de sobrevivência (S e índice 3 de vigor (V] e S. rolfsii em temperatura ambiente (74,4% S e 1 V e c S. sclerotiorum e V. dahliae, ambos em temperatura de freezer (100% S e 3 V. Após um ano de preservação, somente V. dahliae perdeu a patogenicidade na metodologia desenvolvida.Preservation of soilborne phytopathogenic fungi for long periods of time is important so that researches can be followed up at any moment. Soilborne phytopathogenic fungi are organisms that can

  7. Exploring the Potentials of Lysinibacillus sphaericus ZA9 for Plant Growth Promotion and Biocontrol Activities against Phytopathogenic Fungi

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    Zakira Naureen

    2017-08-01

    Full Text Available There is an ongoing hunt for biologically active compounds that can combat phytopathogenic fungi and improve plant growth without causing any hazards to the environment. Consequently the present study aims at deciphering the plant growth promotion and antifungal capability of Lysinibacillus sphaericus ZA9. The bacterium was previously isolated and identified in our laboratory from maize rhizosphere using 16S rRNA gene sequencing. The test bacterium L. sphaericus ZA9 was found to produce high quantity of IAA (697 μg/ mL; siderophores (195.79 μg/ mL, HCN and hydrolytic enzyme as compared to the reference strain Bacillus sphaericus Z2-7. The bacterium was also capable of solubilizing silicates (Si, phosphates (P, and potassium (K. The bacterium enhanced the seedling vigor and germination of seeds pretreated with it and promoted the shoot length of both cucumber and tomato seeds in greenhouse experiment. L. sphaericus ZA9 and its cell free culture supernatant showed varied antagonistic behavior against Alternaria alternata, Curvularia lunata, Aspergillus sp., Sclerotinia sp., Bipolaris spicifera, Trichophyton sp. Fermentation broth culture of L. sphaericus ZA9 was then used to isolate antifungal metabolites by silica column chromatography. Identification and determination of antifungal compounds was carried out by Thin-layer chromatography (TLC followed by NMR spectroscopy. Two compounds were isolated and identified as 2-pentyl-4-quinolinecarboxylic acid (C15H17NO2 which is a quinoline alkaloid and 1- methylcyclohexene which is a cycloalkene. Compound 1; 2-Penthyl-4-quinolinecarboxylic acid was found to be highly antagonistic against most of the fungi tested as compared to the bacterium itself. Its activity was comparable to that of fungicide Benlate, while compound 2; 1- methylcyclohexene did not show any antifungal activity.

  8. THE IMPACT OF FERTILIZATION AND FOLIAR STIMULATION PRODUCTS BOTH ON INCREASING THE RESISTANCE TO MAJOR PHYTOPATHOGENS ATTACKS, AND ON INCREASING THE QUANTITY AND QUALITY OF WINE GRAPES HARVEST

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    Cristina BUNESCU

    2014-12-01

    Full Text Available The paper aimed to demonstrate the impact of fertilization and foliar stimulation products both on increasing the resistance to major phytopathogens attacks, and on increasing the quantity and quality of wine grapes harvest. Applying the foliar fertilizer products Plonvit Kali (c1, Tytanit (c2 and Optysil (c3 to vines, for a period of three years (2011/2013, in phenophases of intensive growth of shoots and grapes at approved dosages, simultaneously with pesticide treatment, not only a reduction of pathogenic fungi attack was obtained, but also and an increase of harvest without diminishing the quality of the grapes.

  9. 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.

  10. 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.

  11. 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.

    Science.gov (United States)

    Aćimović, Srđan G; Zeng, Quan; McGhee, Gayle C; Sundin, George W; Wise, John C

    2015-01-01

    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 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.

  12. 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

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    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.

  13. Criteria for efficient prevention of dissemination and successful eradication of Erwinia amylovora (the cause of fire blight in Aragón, Spain

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    Ana PALACIO-BIELSA

    2013-01-01

    Full Text Available Erwinia amylovora was detected on pome fruits in the Aragón region (North-Eastern Spain, in a ca. 5 km radius area located in the mid Jalón river (mid Ebro Valley in the province of Zaragoza, during 2000‒2003. Eight years have now passed since this pathogen was last detected, without new infections being reported in the same area. The bases for surveys and rapid eradication performed have been analyzed in detail to understand the reasons for the success in removing fireblight. The results demonstrate that intensive surveillance, risk assessment, plant analyses using accurate identification methods, and, especially, rapid total or selective eradication of infected trees in the plots have been very effective in preventing the generalized spread of fireblight and in delaying economic losses associated with this disease. Eradication and compensation to growers, estimated to cost approx. € 467,000, were clearly counterbalanced by the economic value of apple and pear production in the 2000‒2003 period (approx. € 368 million. Fire blight risk-assessment, using the MARYBLYT system, showed that climatic conditions in the studied area were favourable to infections during the analyzed period (1997‒2006. Molecular characterization of E. amylovora strains had revealed their homogeneity, suggesting that these fire blight episodes could have been caused by just one inoculum source, supporting the hypothesis that there was a unique introduction of E. amylovora in the studied area. Spatial spread of E. amylovora to trees was analyzed within six orchards, indicating an aggregated distribution model. This Spanish experience demonstrates the success of scientifically-based prevention methods that lead to the deployment of a fast and strict containment strategy, useful for other Mediterranean areas.

  14. Transfer of several phytopathogenic Pseudomonas species to Acidovorax as Acidovorax avenae subsp. avenae subsp. nov., comb. nov., Acidovorax avenae subsp. citrulli, Acidovorax avenae subsp. cattleyae, and Acidovorax konjaci.

    Science.gov (United States)

    Willems, A; Goor, M; Thielemans, S; Gillis, M; Kersters, K; De Ley, J

    1992-01-01

    DNA-rRNA hybridizations, DNA-DNA hybridizations, polyacrylamide gel electrophoresis of whole-cell proteins, and a numerical analysis of carbon assimilation tests were carried out to determine the relationships among the phylogenetically misnamed phytopathogenic taxa Pseudomonas avenae, Pseudomonas rubrilineans, "Pseudomonas setariae," Pseudomonas cattleyae, Pseudomonas pseudoalcaligenes subsp. citrulli, and Pseudomonas pseudoalcaligenes subsp. konjaci. These organisms are all members of the family Comamonadaceae, within which they constitute a separate rRNA branch. Only P. pseudoalcaligenes subsp. konjaci is situated on the lower part of this rRNA branch; all of the other taxa cluster very closely around the type strain of P. avenae. When they are compared phenotypically, all of the members of this rRNA branch can be differentiated from each other, and they are, as a group, most closely related to the genus Acidovorax. DNA-DNA hybridization experiments showed that these organisms constitute two genotypic groups. We propose that the generically misnamed phytopathogenic Pseudomonas species should be transferred to the genus Acidovorax as Acidovorax avenae and Acidovorax konjaci. Within Acidovorax avenae we distinguished the following three subspecies: Acidovorax avenae subsp. avenae, Acidovorax avenae subsp. cattleyae, and Acidovorax avenae subsp. citrulli. Emended descriptions of the new taxa are presented.

  15. Essential oil composition and antifungal activity of Melissa officinalis originating from north-Est Morocco, against postharvest phytopathogenic fungi in apples.

    Science.gov (United States)

    El Ouadi, Y; Manssouri, M; Bouyanzer, A; Majidi, L; Bendaif, H; Elmsellem, H; Shariati, M A; Melhaoui, A; Hammouti, B

    2017-06-01

    To investigate biological control methods against post-harvest phytopathogenic fungi in apples, tests on the antifungal activity of essential oil of Melissa officinalis were carried out. The essential oil, obtained by hydrodistillation, was analyzed by gas chromatography coupled with mass spectrometry (GC-MS). Analysis of the essential oil was able to detect 88.7% of the components. The main components are P-mentha-1,2,3-triol (13.1%), P-menth-3-en-8-ol (8.8%), pulegone (8.8%), piperitynone oxide (8.4%) and 2-piperitone oxide (7.3%). The determination of the antifungal activity of the essential oil of M. officinalisis carried out in vitro using the technique of poison food (PF) and the volatile activity test (VA). To carry out these two tests, three phytopathogens that cause the deterioration of apples have been selected: Botrytis cinerea, Penicillium expansum and Rhizopus stolonifer. The overall results of this study suggest that M. officinalis essential oil has potential as a bio-antifungal preservative for the control of post-harvest diseases of apple. Copyright © 2017 Elsevier Ltd. All rights reserved.

  16. Genome-wide identification of HrpL-regulated genes in the necrotrophic phytopathogen Dickeya dadantii 3937.

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    Shihui Yang

    Full Text Available BACKGROUND: Dickeya dadantii is a necrotrophic pathogen causing disease in many plants. Previous studies have demonstrated that the type III secretion system (T3SS of D. dadantii is required for full virulence. HrpL is an alternative sigma factor that binds to the hrp box promoter sequence of T3SS genes to up-regulate their expression. METHODOLOGY/PRINCIPAL FINDINGS: To explore the inventory of HrpL-regulated genes of D. dadantii 3937 (3937, transcriptome profiles of wild-type 3937 and a hrpL mutant grown in a T3SS-inducing medium were examined. Using a cut-off value of 1.5, significant differential expression was observed in sixty-three genes, which are involved in various cellular functions such as type III secretion, chemotaxis, metabolism, regulation, and stress response. A hidden Markov model (HMM was used to predict candidate hrp box binding sites in the intergenic regions of 3937, including the promoter regions of HrpL-regulated genes identified in the microarray assay. In contrast to biotrophic phytopathgens such as Pseudomonas syringae, among the HrpL up-regulated genes in 3937 only those within the T3SS were found to contain a hrp box sequence. Moreover, direct binding of purified HrpL protein to the hrp box was demonstrated for hrp box-containing DNA fragments of hrpA and hrpN using the electrophoretic mobility shift assay (EMSA. In this study, a putative T3SS effector DspA/E was also identified as a HrpL-upregulated gene, and shown to be translocated into plant cells in a T3SS-dependent manner. CONCLUSION/SIGNIFICANCES: We provide the genome-wide study of HrpL-regulated genes in a necrotrophic phytopathogen (D. dadantii 3937 through a combination of transcriptomics and bioinformatics, which led to identification of several effectors. Our study indicates the extent of differences for T3SS effector protein inventory requirements between necrotrophic and biotrophic pathogens, and may allow the development of different strategies for

  17. Transposable elements in phytopathogenic Verticillium spp.: insights into genome evolution and inter- and intra-specific diversification

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    Amyotte Stefan G

    2012-07-01

    Full Text Available Abstract Background Verticillium dahliae (Vd and Verticillium albo-atrum (Va are cosmopolitan soil fungi causing very disruptive vascular diseases on a wide range of crop plants. To date, no sexual stage has been identified in either microorganism suggesting that somatic mutation is a major force in generating genetic diversity. Whole genome comparative analysis of the recently sequenced strains VdLs.17 and VaMs.102 revealed that non-random insertions of transposable elements (TEs have contributed to the generation of four lineage-specific (LS regions in VdLs.17. Results We present here a detailed analysis of Class I retrotransposons and Class II “cut-and-paste” DNA elements detected in the sequenced Verticillium genomes. We report also of their distribution in other Vd and Va isolates from various geographic origins. In VdLs.17, we identified and characterized 56 complete retrotransposons of the Gypsy-, Copia- and LINE-like types, as well as 34 full-length elements of the “cut-and-paste” superfamilies Tc1/mariner, Activator and Mutator. While Copia and Tc1/mariner were present in multiple identical copies, Activator and Mutator sequences were highly divergent. Most elements comprised complete ORFs, had matching ESTs and showed active transcription in response to stress treatment. Noticeably, we found evidences of repeat-induced point mutation (RIP only in some of the Gypsy retroelements. While Copia-, Gypsy- and Tc1/mariner-like transposons were prominent, a large variation in presence of the other types of mobile elements was detected in the other Verticillium spp. strains surveyed. In particular, neither complete nor defective “cut-and-paste” TEs were found in VaMs.102. Conclusions Copia-, Gypsy- and Tc1/mariner-like transposons are the most wide-spread TEs in the phytopathogens V. dahliae and V. albo-atrum. In VdLs.17, we identified several retroelements and “cut-and-paste” transposons still potentially active. Some of these

  18. Anti-phytopathogen potential of endophytic actinobacteria isolated from tomato plants (Lycopersicon esculentum) in southern Brazil, and characterization of Streptomyces sp. R18(6), a potential biocontrol agent.

    Science.gov (United States)

    de Oliveira, Margaroni Fialho; da Silva, Mariana Germano; Van Der Sand, Sueli T

    2010-09-01

    Tomato plants (Lycopersicon esculentum) are highly susceptible to phytopathogen attack. The resulting intensive application of pesticides on tomato crops can affect the environment and health of humans and animals. The objective of this study was to select potential biocontrol agents among actinobacteria from tomato plants, in a search for alternative phytopathogen control. We evaluated 70 endophytic actinobacteria isolated from tomato plants in southern Brazil, testing their antimicrobial activity, siderophore production, indoleacetic acid production, and phosphate solubility. The actinomycete isolate with the highest antimicrobial potential was selected using the agar-well diffusion method, in order to optimize conditions for the production of compounds with antimicrobial activity. For this study, six growth media (starch casein-SC, ISP2, Bennett's, Sahin, Czapek-Dox, and TSB), three temperatures (25 degrees C, 30 degrees C, and 35 degrees C) and different pH were tested. Of the actinobacteria tested, 88.6% showed antimicrobial activity against at least one phytopathogen, 72.1% showed a positive reaction for indoleacetic acid production, 86.8% produced siderophores and 16.2% showed a positive reaction for phosphate solubility. Isolate R18(6) was selected due to its antagonistic activity against all phytopathogenic microorganisms tested in this study. The best conditions for production were observed in the SC medium, at 30 degrees C and pH 7.0. The isolate R18(6) showed close biochemical and genetic similarity to Streptomyces pluricolorescens. Copyright 2010 Elsevier Masson SAS. All rights reserved.

  19. kdgREcc Negatively Regulates Genes for Pectinases, Cellulase, Protease, HarpinEcc, and a Global RNA Regulator in Erwinia carotovora subsp. carotovora†

    Science.gov (United States)

    Liu, Yang; Jiang, Guoqiao; Cui, Yaya; Mukherjee, Asita; Ma, Wei Lei; Chatterjee, Arun K.

    1999-01-01

    Erwinia carotovora subsp. carotovora produces extracellular pectate lyase (Pel), polygalacturonase (Peh), cellulase (Cel), and protease (Prt). The concerted actions of these enzymes largely determine the virulence of this plant-pathogenic bacterium. E. carotovora subsp. carotovora also produces HarpinEcc, the elicitor of the hypersensitive reaction. We document here that KdgREcc (Kdg, 2-keto-3-deoxygluconate; KdgR, general repressor of genes involved in pectin and galacturonate catabolism), a homolog of the E. chrysanthemi repressor, KdgREch and the Escherichia coli repressor, KdgREco, negatively controls not only the pectinases, Pel and Peh, but also Cel, Prt, and HarpinEcc production in E. carotovora subsp. carotovora. The levels of pel-1, peh-1, celV, and hrpNEcc transcripts are markedly affected by KdgREcc. The KdgREcc− mutant is more virulent than the KdgREcc+ parent. Thus, our data for the first time establish a global regulatory role for KdgREcc in E. carotovora subsp. carotovora. Another novel observation is the negative effect of KdgREcc on the transcription of rsmB (previously aepH), which specifies an RNA regulator controlling exoenzyme and HarpinEcc production. The levels of rsmB RNA are higher in the KdgREcc− mutant than in the KdgREcc+ parent. Moreover, by DNase I protection assays we determined that purified KdgREcc protected three 25-bp regions within the transcriptional unit of rsmB. Alignment of the protected sequences revealed the 21-mer consensus sequence of the KdgREcc-binding site as 5′-G/AA/TA/TGAAA[N6]TTTCAG/TG/TA-3′. Two such KdgREcc-binding sites occur in rsmB DNA in a close proximity to each other within nucleotides +79 and +139 and the third KdgREcc-binding site within nucleotides +207 and +231. Analysis of lacZ transcriptional fusions shows that the KdgR-binding sites negatively affect the expression of rsmB. KdgREcc also binds the operator DNAs of pel-1 and peh-1 genes and represses expression of a pel1-lacZ and a peh1-lac

  20. kdgREcc negatively regulates genes for pectinases, cellulase, protease, HarpinEcc, and a global RNA regulator in Erwinia carotovora subsp. carotovora.

    Science.gov (United States)

    Liu, Y; Jiang, G; Cui, Y; Mukherjee, A; Ma, W L; Chatterjee, A K

    1999-04-01

    Erwinia carotovora subsp. carotovora produces extracellular pectate lyase (Pel), polygalacturonase (Peh), cellulase (Cel), and protease (Prt). The concerted actions of these enzymes largely determine the virulence of this plant-pathogenic bacterium. E. carotovora subsp. carotovora also produces HarpinEcc, the elicitor of the hypersensitive reaction. We document here that KdgREcc (Kdg, 2-keto-3-deoxygluconate; KdgR, general repressor of genes involved in pectin and galacturonate catabolism), a homolog of the E. chrysanthemi repressor, KdgREch and the Escherichia coli repressor, KdgREco, negatively controls not only the pectinases, Pel and Peh, but also Cel, Prt, and HarpinEcc production in E. carotovora subsp. carotovora. The levels of pel-1, peh-1, celV, and hrpNEcc transcripts are markedly affected by KdgREcc. The KdgREcc- mutant is more virulent than the KdgREcc+ parent. Thus, our data for the first time establish a global regulatory role for KdgREcc in E. carotovora subsp. carotovora. Another novel observation is the negative effect of KdgREcc on the transcription of rsmB (previously aepH), which specifies an RNA regulator controlling exoenzyme and HarpinEcc production. The levels of rsmB RNA are higher in the KdgREcc- mutant than in the KdgREcc+ parent. Moreover, by DNase I protection assays we determined that purified KdgREcc protected three 25-bp regions within the transcriptional unit of rsmB. Alignment of the protected sequences revealed the 21-mer consensus sequence of the KdgREcc-binding site as 5'-G/AA/TA/TGAAA[N6]TTTCAG/TG/TA-3'. Two such KdgREcc-binding sites occur in rsmB DNA in a close proximity to each other within nucleotides +79 and +139 and the third KdgREcc-binding site within nucleotides +207 and +231. Analysis of lacZ transcriptional fusions shows that the KdgR-binding sites negatively affect the expression of rsmB. KdgREcc also binds the operator DNAs of pel-1 and peh-1 genes and represses expression of a pel1-lacZ and a peh1-lac

  1. Structural and functional characterization of the GalNAc/Gal-specific lectin from the phytopathogenic ascomycete Sclerotinia sclerotiorum (Lib.) de Bary.

    Science.gov (United States)

    Candy, Laure; Van Damme, Els J M; Peumans, Willy J; Menu-Bouaouiche, Laurence; Erard, Monique; Rougé, Pierre

    2003-08-22

    The lectin found in mycelium and sclerotes of the phytopathogenic fungus Sclerotinia sclerotiorum is a homodimer consisting of two identical non-covalently bound subunits of 16,000 Da. CD spectra analysis revealed that the S. sclerotiorum agglutinin (SSA) contains predominantly beta-sheet structures. SSA exhibits specificity towards GalNAc whereby the hydroxyls at positions 4 and 6 of the pyranose ring play a key role in the interaction with simple sugars. The carbohydrate-binding site of SSA can also accommodate disaccharides. The N-terminal sequence of SSA shares no significant similarity with any other protein except a lectin from the Sclerotiniaceae species Ciborinia camelliae. A comparison of SSA and the lectins from C. camelliae and some previously characterized lectins indicates that the Sclerotiniaceae lectins form a homogeneous family of fungal lectins. This newly identified lectin family, which is structurally unrelated to any other family of fungal lectins, is most probably confined to the Ascomycota.

  2. Effect of crude plant extracts from some Oaxacan flora on two deleterious fungal phytopathogens and extract compatibility with a biofertilizer strain.

    Science.gov (United States)

    Lira-De León, Karla I; Ramírez-Mares, Marco V; Sánchez-López, Vladimir; Ramírez-Lepe, Mario; Salas-Coronado, Raúl; Santos-Sánchez, Norma F; Valadez-Blanco, Rogelio; Hernández-Carlos, Beatriz

    2014-01-01

    The antimicrobial activity of 12 plant extracts was tested against the phytopathogens Alternaria alternata and Fusarium solani. In addition, the compatibility of the extracts toward Bacillus liqueniformis, a biofertilizer and a non-target microorganism, was assessed. Plants tested belong to the Euphorbiaceae, Asteraceae, Crassulaceae, Rubiaceae, Convolvulaceae, Verbenaceae, Orchidaceae, Nyctaginaceae, Boraginaceae, and Tiliaceae families and were collected in the State of Oaxaca. The antifungal activity of the plant extracts (50-100 mg/mL) against A. alternata and F. solani, was determined by measuring the mycelium radial growth and obtaining the minimum inhibitory concentration (MIC) of fungal growth. In addition, with the aim of finding plant extracts which are compatible with a B. licheniformis biofertilizer strain and to test the non-toxic nature of the treatments, the toxicity of the extracts toward this strain was evaluated using the agar diffusion method. Azoxystrobin (12 μg) and chloramphenicol (30 μg) were used as positive controls for the pathogens and for the non-target bacteria, respectively. Plant extracts inhibited fungal growth in the ranges of 0.76-56.17% against F. solani and 2.02-69.07% against A. alternata. The extracts of Acalypha subviscida, Ipomoea murucoides, Tournefortia densiflora and Lantana achyranthifolia showed MIC values between 5.77-12.5 mg/mL for at least one of the fungal species. The best treatment, Adenophyllum aurantium, exhibited a maximum inhibition for both F. solani (56.17%, MIC = 7.78 mg/mL) and A. alternata (68.64% MIC = 7.78 mg/mL), and resulted innocuous toward B. licheniformis. Therefore, this plant has an outstanding potential for the agroecological control of fungal phytopathogens in industrial crops.

  3. Transcriptomic profiling of microbe-microbe interactions reveals the specific response of the biocontrol strain P. fluorescens In5 to the phytopathogen Rhizoctonia solani

    DEFF Research Database (Denmark)

    Hennessy, Rosanna Catherine; Glaring, Mikkel Andreas; Olsson, Stefan

    2017-01-01

    metabolites, e.g. non-ribosomal synthetases and hydrogen cyanide were not differentially expressed at the time points studied. CONCLUSION: This study demonstrates that genes possibly involved in metabolite detoxification are highly upregulated in P. fluorescens In5 when co-cultured with plant pathogens......BACKGROUND: Few studies to date report the transcriptional response of biocontrol bacteria toward phytopathogens. In order to gain insights into the potential mechanism underlying the antagonism of the antimicrobial producing strain P. fluorescens In5 against the phytopathogens Rhizoctonia solani....... fluorescens In5 or bacterial cultures in dual-culture for 48 h with each pathogen in biological triplicates. RNA-seq libraries were constructed following a default Illumina stranded RNA protocol including rRNA depletion and were sequenced 2 × 100 bases on Illumina HiSeq generating approximately 10 million...

  4. Transcriptomic profiling of microbe-microbe interactions reveals the specific response of the biocontrol strain P. fluorescens In5 to the phytopathogen Rhizoctonia solani.

    Science.gov (United States)

    Hennessy, Rosanna C; Glaring, Mikkel A; Olsson, Stefan; Stougaard, Peter

    2017-08-10

    Few studies to date report the transcriptional response of biocontrol bacteria toward phytopathogens. In order to gain insights into the potential mechanism underlying the antagonism of the antimicrobial producing strain P. fluorescens In5 against the phytopathogens Rhizoctonia solani and Pythium aphanidermatum, global RNA sequencing was performed. Differential gene expression profiling of P. fluorescens In5 in response to either R. solani or P. aphanidermatum was investigated using transcriptome sequencing (RNA-seq). Total RNA was isolated from single bacterial cultures of P. fluorescens In5 or bacterial cultures in dual-culture for 48 h with each pathogen in biological triplicates. RNA-seq libraries were constructed following a default Illumina stranded RNA protocol including rRNA depletion and were sequenced 2 × 100 bases on Illumina HiSeq generating approximately 10 million reads per sample. No significant changes in global gene expression were recorded during dual-culture of P. fluorescens In5 with any of the two pathogens but rather each pathogen appeared to induce expression of a specific set of genes. A particularly strong transcriptional response to R. solani was observed and notably several genes possibly associated with secondary metabolite detoxification and metabolism were highly upregulated in response to the fungus. A total of 23 genes were significantly upregulated and seven genes were significantly downregulated with at least respectively a threefold change in expression level in response to R. solani compared to the no fungus control. In contrast, only one gene was significantly upregulated over threefold and three transcripts were significantly downregulated over threefold in response to P. aphanidermatum. Genes known to be involved in synthesis of secondary metabolites, e.g. non-ribosomal synthetases and hydrogen cyanide were not differentially expressed at the time points studied. This study demonstrates that genes possibly involved in

  5. Molecular basis of arabinobio-hydrolase activity in phytopathogenic fungi: crystal structure and catalytic mechanism of Fusarium graminearum GH93 exo-alpha-L-arabinanase.

    Science.gov (United States)

    Carapito, Raphaël; Imberty, Anne; Jeltsch, Jean-Marc; Byrns, Simon C; Tam, Pui-Hang; Lowary, Todd L; Varrot, Annabelle; Phalip, Vincent

    2009-05-01

    The phytopathogenic fungus Fusarium graminearum secretes a very diverse pool of glycoside hydrolases (GHs) aimed at degrading plant cell walls. alpha-l-Arabinanases are essential GHs participating in the complete hydrolysis of hemicellulose, a natural resource for various industrial processes, such as bioethanol or pharmaceuticals production. Arb93A, the exo-1,5-alpha-l-arabinanase of F. graminearum encoded by the gene fg03054.1, belongs to the GH93 family, for which no structural data exists. The enzyme is highly active (1065 units/mg) and displays a strict substrate specificity for linear alpha-1,5-l-arabinan. Biochemical assays and NMR experiments demonstrated that the enzyme releases alpha-1,5-l-arabinobiose from the nonreducing end of the polysaccharide. We determined the crystal structure of the native enzyme and its complex with alpha-1,5-l-arabinobiose, a degradation product of alpha-Me-1,5-l-arabinotetraose, at 1.85 and 2.05A resolution, respectively. Arb93A is a monomeric enzyme, which presents the six-bladed beta-propeller fold characteristic of sialidases of clan GHE. The configuration of the bound arabinobiose is consistent with the retaining mechanism proposed for the GH93 family. Catalytic residues were proposed from the structural analysis, and site-directed mutagenesis was used to validate their role. They are significantly different from those observed for GHE sialidases.

  6. Enrichment of chitinolytic microorganisms: isolation and characterization of a chitinase exhibiting antifungal activity against phytopathogenic fungi from a novel Streptomyces strain.

    Science.gov (United States)

    Hoster, Frank; Schmitz, Jessica E; Daniel, Rolf

    2005-01-01

    Thirteen different chitin-degrading bacteria were isolated from soil and sediment samples. Five of these strains (SGE2, SGE4, SSL3, MG1, and MG3) exhibited antifungal activity against phytopathogenic fungi. Analyses of the 16S rRNA genes and the substrate spectra revealed that the isolates belong to the genera Bacillus or Streptomyces. The closest relatives were Bacillus chitinolyticus (SGE2, SGE4, and SSL3), B. ehimensis (MG1), and Streptomyces griseus (MG3). The chitinases present in the culture supernatants of the five isolates revealed optimal activity between 45 degrees C and 50 degrees C and at pH values of 4 (SSL3), 5 (SGE2 and MG1), 6 (SGE4), and 5-7 (MG3). The crude chitinase preparations of all five strains possessed antifungal activity. The chitinase of MG3 (ChiIS) was studied further, since the crude enzyme conferred strong growth suppression of all fungi tested and was very active over the entire pH range tested. The chiIS gene was cloned and the gene product was purified. The deduced protein consisted of 303 amino acids with a predicted molecular mass of 31,836 Da. Sequence analysis revealed that ChiIS of MG3 is similar to chitinases of Streptomyces species, which belong to family 19 of glycosyl hydrolases. Purified ChiIS showed remarkable antifungal activity and stability.

  7. Potential of biosynthesized silver nanoparticles using Stenotrophomonas sp. BHU-S7 (MTCC 5978) for management of soil-borne and foliar phytopathogens.

    Science.gov (United States)

    Mishra, Sandhya; Singh, Braj Raj; Naqvi, Alim H; Singh, H B

    2017-03-27

    Stenotrophomonas sp. is emerging as a popular microbe of global concern with various potential ecological roles. Biosynthesis of gold and silver nanoparticles (AgNPs) using this bacterial strain has shown promising applications in life sciences. However, there is no report on efficient agricultural applications of biosynthesized AgNPs using Stenotrophomonas sp. In this regard, successful biosynthesis of AgNPs using Stenotrophomonas sp. BHU-S7 (MTCC 5978) was monitored by Uv-visible spectrum showing surface plasmon resonance (SPR) peak at 440 nm. The biosynthesized AgNPs were spherical with an average mean size of ~12 nm. The antifungal efficacy of biosynthesized AgNPs against foliar and soil-borne phytopathogens was observed. The inhibitory impact of AgNPs (2, 4, 10 μg/ml) on conidial germination was recorded under in vitro conditions. Interestingly, sclerotia of Sclerotium rolfsii exposed to AgNPs failed to germinate on PDA medium and in soil system. Moreover, AgNPs treatment successfully managed collar rot of chickpea caused by S. rolfsii under greenhouse conditions. The reduced sclerotia germination, phenolic acids induction, altered lignification and H 2 O 2 production was observed to be the probable mechanisms providing protection to chickpea against S. rolfsii. Our data revealed that AgNPs treated plants are better equipped to cope with pathogen challenge pointing towards their robust applications in plant disease management.

  8. FTIR spectroscopic evaluation of changes in the cellular biochemical composition of the phytopathogenic fungus Alternaria alternata induced by extracts of some Greek medicinal and aromatic plants

    Science.gov (United States)

    Skotti, Efstathia; Kountouri, Sophia; Bouchagier, Pavlos; Tsitsigiannis, Dimitrios I.; Polissiou, Moschos; Tarantilis, Petros A.

    2014-06-01

    In this study, the biological activity of aquatic extracts of selected Greek medicinal and aromatic plants to the phytopathogenic fungus Alternaria alternata was investigated. Lamiaceae species (Hyssopus officinalis L., Melissa officinalis L., Origanum dictamnus L., Origanum vulgare L. and Salvia officinalis L.) were found to enhance significantly the mycelium growth whereas Crocus sativus appears to inhibit it slightly. M. officinalis and S. officinalis caused the highest stimulation in mycelium growth (+97%) and conidia production (+65%) respectively. In order to further investigate the bioactivity of plant extracts to A. alternata, we employed Fourier Transform Infrared Spectroscopy (FTIR). Differences of original spectra were assigned mainly to amides of proteins. The second derivative transformation of spectra revealed changes in spectral regions corresponding to absorptions of the major cellular constituents such as cell membrane and proteins. Principal component analysis of the second derivative transformed spectra confirmed that fatty acids of the cell membranes, amides of proteins and polysaccharides of the cell wall had the major contribution to data variation. FTIR band area ratios were found to correlate with fungal mycelium growth.

  9. Strigolactone biosynthesis is evolutionarily conserved, regulated by phosphate starvation and contributes to resistance against phytopathogenic fungi in a moss, Physcomitrella patens

    KAUST Repository

    Decker, Eva L.

    2017-03-06

    In seed plants, strigolactones (SLs) regulate architecture and induce mycorrhizal symbiosis in response to environmental cues. SLs are formed by combined activity of the carotenoid cleavage dioxygenases (CCDs) 7 and 8 from 9-cis-β-carotene, leading to carlactone that is converted by cytochromes P450 (clade 711; MAX1 in Arabidopsis) into various SLs. As Physcomitrella patens possesses CCD7 and CCD8 homologs but lacks MAX1, we investigated if PpCCD7 together with PpCCD8 form carlactone and how deletion of these enzymes influences growth and interactions with the environment. We investigated the enzymatic activity of PpCCD7 and PpCCD8 in vitro, identified the formed products by high performance liquid chromatography (HPLC) and LC-MS, and generated and analysed ΔCCD7 and ΔCCD8 mutants. We defined enzymatic activity of PpCCD7 as a stereospecific 9-cis-CCD and PpCCD8 as a carlactone synthase. ΔCCD7 and ΔCCD8 lines showed enhanced caulonema growth, which was revertible by adding the SL analogue GR24 or carlactone. Wild-type (WT) exudates induced seed germination in Orobanche ramosa. This activity was increased upon phosphate starvation and abolished in exudates of both mutants. Furthermore, both mutants showed increased susceptibility to phytopathogenic fungi. Our study reveals the deep evolutionary conservation of SL biosynthesis, SL function, and its regulation by biotic and abiotic cues.

  10. Fatty acid DSF binds and allosterically activates histidine kinase RpfC of phytopathogenic bacterium Xanthomonas campestris pv. campestris to regulate quorum-sensing and virulence.

    Directory of Open Access Journals (Sweden)

    Zhen Cai

    2017-04-01

    Full Text Available As well as their importance to nutrition, fatty acids (FA represent a unique group of quorum sensing chemicals that modulate the behavior of bacterial population in virulence. However, the way in which full-length, membrane-bound receptors biochemically detect FA remains unclear. Here, we provide genetic, enzymological and biophysical evidences to demonstrate that in the phytopathogenic bacterium Xanthomonas campestris pv. campestris, a medium-chain FA diffusible signal factor (DSF binds directly to the N-terminal, 22 amino acid-length sensor region of a receptor histidine kinase (HK, RpfC. The binding event remarkably activates RpfC autokinase activity by causing an allosteric change associated with the dimerization and histidine phosphotransfer (DHp and catalytic ATP-binding (CA domains. Six residues were found essential for sensing DSF, especially those located in the region adjoining to the inner membrane of cells. Disrupting direct DSF-RpfC interaction caused deficiency in bacterial virulence and biofilm development. In addition, two amino acids within the juxtamembrane domain of RpfC, Leu172 and Ala178, are involved in the autoinhibition of the RpfC kinase activity. Replacements of them caused constitutive activation of RpfC-mediated signaling regardless of DSF stimulation. Therefore, our results revealed a biochemical mechanism whereby FA activates bacterial HK in an allosteric manner, which will assist in future studies on the specificity of FA-HK recognition during bacterial virulence regulation and cell-cell communication.

  11. Green synthesis of protein capped silver nanoparticles from phytopathogenic fungus Macrophomina phaseolina (Tassi) Goid with antimicrobial properties against multidrug-resistant bacteria

    Science.gov (United States)

    2014-01-01

    In recent years, green synthesis of nanoparticles, i.e., synthesizing nanoparticles using biological sources like bacteria, algae, fungus, or plant extracts have attracted much attention due to its environment-friendly and economic aspects. The present study demonstrates an eco-friendly and low-cost method of biosynthesis of silver nanoparticles using cell-free filtrate of phytopathogenic fungus Macrophomina phaseolina. UV-visible spectrum showed a peak at 450 nm corresponding to the plasmon absorbance of silver nanoparticles. Scanning electron microscopy (SEM), atomic force microscopy (AFM), and transmission electron microscopy (TEM) revealed the presence of spherical silver nanoparticles of the size range 5 to 40 nm, most of these being 16 to 20 nm in diameter. X-ray diffraction (XRD) spectrum of the nanoparticles exhibited 2θ values corresponding to silver nanoparticles. These nanoparticles were found to be naturally protein coated. SDS-PAGE analysis showed the presence of an 85-kDa protein band responsible for capping and stabilization of the silver nanoparticles. Antimicrobial activities of the silver nanoparticles against human as well as plant pathogenic multidrug-resistant bacteria were assayed. The particles showed inhibitory effect on the growth kinetics of human and plant bacteria. Furthermore, the genotoxic potential of the silver nanoparticles with increasing concentrations was evaluated by DNA fragmentation studies using plasmid DNA. PMID:25114655

  12. Green synthesis of protein capped silver nanoparticles from phytopathogenic fungus Macrophomina phaseolina (Tassi) Goid with antimicrobial properties against multidrug-resistant bacteria

    Science.gov (United States)

    Chowdhury, Supriyo; Basu, Arpita; Kundu, Surekha

    2014-07-01

    In recent years, green synthesis of nanoparticles, i.e., synthesizing nanoparticles using biological sources like bacteria, algae, fungus, or plant extracts have attracted much attention due to its environment-friendly and economic aspects. The present study demonstrates an eco-friendly and low-cost method of biosynthesis of silver nanoparticles using cell-free filtrate of phytopathogenic fungus Macrophomina phaseolina. UV-visible spectrum showed a peak at 450 nm corresponding to the plasmon absorbance of silver nanoparticles. Scanning electron microscopy (SEM), atomic force microscopy (AFM), and transmission electron microscopy (TEM) revealed the presence of spherical silver nanoparticles of the size range 5 to 40 nm, most of these being 16 to 20 nm in diameter. X-ray diffraction (XRD) spectrum of the nanoparticles exhibited 2 θ values corresponding to silver nanoparticles. These nanoparticles were found to be naturally protein coated. SDS-PAGE analysis showed the presence of an 85-kDa protein band responsible for capping and stabilization of the silver nanoparticles. Antimicrobial activities of the silver nanoparticles against human as well as plant pathogenic multidrug-resistant bacteria were assayed. The particles showed inhibitory effect on the growth kinetics of human and plant bacteria. Furthermore, the genotoxic potential of the silver nanoparticles with increasing concentrations was evaluated by DNA fragmentation studies using plasmid DNA.

  13. Engineering of Bacteriophages Y2::dpoL1-C and Y2::luxAB for Efficient Control and Rapid Detection of the Fire Blight Pathogen, Erwinia amylovora.

    Science.gov (United States)

    Born, Yannick; Fieseler, Lars; Thöny, Valentin; Leimer, Nadja; Duffy, Brion; Loessner, Martin J

    2017-06-15

    Erwinia amylovora is the causative agent of fire blight, a devastating plant disease affecting members of the Rosaceae Alternatives to antibiotics for control of fire blight symptoms and outbreaks are highly desirable, due to increasing drug resistance and tight regulatory restrictions. Moreover, the available diagnostic methods either lack sensitivity, lack speed, or are unable to discriminate between live and dead bacteria. Owing to their extreme biological specificity, bacteriophages are promising alternatives for both aims. In this study, the virulent broad-host-range E. amylovora virus Y2 was engineered to enhance its killing activity and for use as a luciferase reporter phage, respectively. Toward these aims, a depolymerase gene of E. amylovora virus L1 ( dpoL1-C ) or a bacterial luxAB fusion was introduced into the genome of Y2 by homologous recombination. The genes were placed downstream of the major capsid protein orf68 , under the control of the native promoter. The modifications did not affect viability of infectivity of the recombinant viruses. Phage Y2:: dpoL1-C demonstrated synergistic activity between the depolymerase degrading the exopolysaccharide capsule and phage infection, which greatly enhanced bacterial killing. It also significantly reduced the ability of E. amylovora to colonize the surface of detached flowers. The reporter phage Y2:: luxAB transduced bacterial luciferase into host cells and induced synthesis of large amounts of a LuxAB luciferase fusion. After the addition of aldehyde substrate, bioluminescence could be readily monitored, and this enabled rapid and specific detection of low numbers of viable bacteria, without enrichment, both in vitro and in plant material. IMPORTANCE Fire blight, caused by Erwinia amylovora , is the major threat to global pome fruit production, with high economic losses every year. Bacteriophages represent promising alternatives to not only control the disease, but also for rapid diagnostics. To enhance

  14. The crystal structure of Erwinia amylovora AmyR, a member of the YbjN protein family, shows similarity to type III secretion chaperones but suggests different cellular functions.

    Science.gov (United States)

    Bartho, Joseph D; Bellini, Dom; Wuerges, Jochen; Demitri, Nicola; Toccafondi, Mirco; Schmitt, Armin O; Zhao, Youfu; Walsh, Martin A; Benini, Stefano

    2017-01-01

    AmyR is a stress and virulence associated protein from the plant pathogenic Enterobacteriaceae species Erwinia amylovora, and is a functionally conserved ortholog of YbjN from Escherichia coli. The crystal structure of E. amylovora AmyR reveals a class I type III secretion chaperone-like fold, despite the lack of sequence similarity between these two classes of protein and lacking any evidence of a secretion-associated role. The results indicate that AmyR, and YbjN proteins in general, function through protein-protein interactions without any enzymatic action. The YbjN proteins of Enterobacteriaceae show remarkably low sequence similarity with other members of the YbjN protein family in Eubacteria, yet a high level of structural conservation is observed. Across the YbjN protein family sequence conservation is limited to residues stabilising the protein core and dimerization interface, while interacting regions are only conserved between closely related species. This study presents the first structure of a YbjN protein from Enterobacteriaceae, the most highly divergent and well-studied subgroup of YbjN proteins, and an in-depth sequence and structural analysis of this important but poorly understood protein family.

  15. MRN- and 9-1-1-Independent Activation of the ATR-Chk1 Pathway during the Induction of the Virulence Program in the Phytopathogen Ustilago maydis.

    Directory of Open Access Journals (Sweden)

    María Tenorio-Gómez

    Full Text Available DNA damage response (DDR leads to DNA repair, and depending on the extent of the damage, to further events, including cell death. Evidence suggests that cell differentiation may also be a consequence of the DDR. During the formation of the infective hypha in the phytopathogenic fungus Ustilago maydis, two DDR kinases, Atr1 and Chk1, are required to induce a G2 cell cycle arrest, which in turn is essential to display the virulence program. However, the triggering factor of DDR in this process has remained elusive. In this report we provide data suggesting that no DNA damage is associated with the activation of the DDR during the formation of the infective filament in U. maydis. We have analyzed bulk DNA replication during the formation of the infective filament, and we found no signs of impaired DNA replication. Furthermore, using RPA-GFP fusion as a surrogate marker of the presence of DNA damage, we were unable to detect any sign of DNA damage at the cellular level. In addition, neither MRN nor 9-1-1 complexes, both instrumental to transmit the DNA damage signal, are required for the induction of the above mentioned cell cycle arrest, as well as for virulence. In contrast, we have found that the claspin-like protein Mrc1, which in other systems serves as scaffold for Atr1 and Chk1, was required for both processes. We discuss possible alternative ways to trigger the DDR, independent of DNA damage, in U. maydis during virulence program activation.

  16. A novel single-stranded RNA virus isolated from a phytopathogenic filamentous fungus, Rosellinia necatrix, with similarity to hypo-like viruses

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    Rui eZhang

    2014-07-01

    Full Text Available Here we report a biological and molecular characterization of a novel positive-sense RNA virus isolated from a field isolate (NW10 of a filamentous phytopathogenic fungus, the white root rot fungus that is designated as Rosellinia necatrix fusarivirus 1 (RnFV1. A recently developed technology using zinc ions allowed us to transfer RnFV1 to two mycelially incompatible Rosellinia necatrix strains. A biological comparison of the virus-free and -recipient isogenic fungal strains suggested that RnFV1 infects latently and thus has no potential as a virocontrol agent. The virus has an undivided positive-sense RNA genome of 6286 nucleotides excluding a poly (A tail. The genome possesses two non-overlapping open reading frames (ORFs: a large ORF1 that encodes polypeptides with RNA replication functions and a smaller ORF2 that encodes polypeptides of unknown function. A lack of coat protein genes was suggested by the failure of virus particles from infected mycelia. No evidence was obtained by Northern analysis or classical 5'-RACE for the presence of subgenomic RNA for the downstream ORF. Sequence similarities were found in amino-acid sequence between RnFV1 putative proteins and counterparts of a previously reported mycovirus, Fusarium graminearum virus 1 (FgV1. Interestingly, several related sequences were detected by BLAST searches of independent transcriptome assembly databases one of which probably represents an entire virus genome. Phylogenetic analysis based on the conserved RNA-dependent RNA polymerase showed that RnFV1, FgV1, and these similar sequences are grouped in a cluster distinct from distantly related hypoviruses. It is proposed that a new taxonomic family termed Fusariviridae be created to include RnFV1and FgV1.

  17. Strigolactone biosynthesis is evolutionarily conserved, regulated by phosphate starvation and contributes to resistance against phytopathogenic fungi in a moss, Physcomitrella patens.

    Science.gov (United States)

    Decker, Eva L; Alder, Adrian; Hunn, Stefan; Ferguson, Jenny; Lehtonen, Mikko T; Scheler, Bjoern; Kerres, Klaus L; Wiedemann, Gertrud; Safavi-Rizi, Vajiheh; Nordzieke, Steffen; Balakrishna, Aparna; Baz, Lina; Avalos, Javier; Valkonen, Jari P T; Reski, Ralf; Al-Babili, Salim

    2017-10-01

    In seed plants, strigolactones (SLs) regulate architecture and induce mycorrhizal symbiosis in response to environmental cues. SLs are formed by combined activity of the carotenoid cleavage dioxygenases (CCDs) 7 and 8 from 9-cis-β-carotene, leading to carlactone that is converted by cytochromes P450 (clade 711; MAX1 in Arabidopsis) into various SLs. As Physcomitrella patens possesses CCD7 and CCD8 homologs but lacks MAX1, we investigated if PpCCD7 together with PpCCD8 form carlactone and how deletion of these enzymes influences growth and interactions with the environment. We investigated the enzymatic activity of PpCCD7 and PpCCD8 in vitro, identified the formed products by high performance liquid chromatography (HPLC) and LC-MS, and generated and analysed ΔCCD7 and ΔCCD8 mutants. We defined enzymatic activity of PpCCD7 as a stereospecific 9-cis-CCD and PpCCD8 as a carlactone synthase. ΔCCD7 and ΔCCD8 lines showed enhanced caulonema growth, which was revertible by adding the SL analogue GR24 or carlactone. Wild-type (WT) exudates induced seed germination in Orobanche ramosa. This activity was increased upon phosphate starvation and abolished in exudates of both mutants. Furthermore, both mutants showed increased susceptibility to phytopathogenic fungi. Our study reveals the deep evolutionary conservation of SL biosynthesis, SL function, and its regulation by biotic and abiotic cues. © 2017 The Authors. New Phytologist © 2017 New Phytologist Trust.

  18. Bio-fabrication of silver nanoparticles using the leaf extract of an ancient herbal medicine, dandelion (Taraxacum officinale), evaluation of their antioxidant, anticancer potential, and antimicrobial activity against phytopathogens.

    Science.gov (United States)

    Saratale, Rijuta G; Benelli, Giovanni; Kumar, Gopalakrishnan; Kim, Dong Su; Saratale, Ganesh D

    2018-04-01

    In recent years, the use of nanoparticle-based antimicrobials has been increased due to many advantages over conventional agrochemicals. This study investigates the utilization of common medicinal plant dandelion, Taraxacum officinale, for the synthesis of silver nanoparticles (TOL-AgNPs). AgNPs were evaluated for antimicrobial activity against two important phytopathogens, Xanthomonas axonopodis and Pseudomonas syringae. The morphology, size, and structure of TOL-AgNPs were characterized using UV-visible spectroscopy and X-ray diffraction (XRD). Fourier transform infrared spectroscopy (FT-IR) showed the presence of phytochemicals involved during synthesis of NPs. High-resolution transmission electron microscopy (HR-TEM) analysis shed light on the size of monodispersed spherical AgNPs ranging between 5 and 30 nm, with an average particle size of about 15 nm. The TOL-AgNPs (at 20 μg/mL concentration) showed significant antibacterial activity with significant growth inhibition of phytopathogens X. axonopodis (22.0 ± 0.84 mm) and P. syringae (19.5 ± 0.66 mm). The synthesized AgNPs had higher antibacterial activity in comparison with commercial AgNPs. Synergistic assays with standard antibiotics revealed that nanoformulations with tetracycline showed better broad-spectrum efficiency to control phytopathogens. They also possessed significant antioxidant potential in terms of ABTS (IC 50  = 45.6 μg/mL), DPPH (IC 50  = 56.1 μg/mL), and NO (IC 50  = 55.2 μg/mL) free radical scavenging activity. The TOL-AgNPs showed high cytotoxic effect against human liver cancer cells (HepG2). Overall, dandelion-mediated AgNPs synthesis can represent a novel approach to develop effective antimicrobial and anticancer drugs with a cheap and eco-friendly nature.

  19. A facile and efficient synthesis of diaryl amines or ethers under microwave irradiation at presence of KF/Al2O3 without solvent and their anti-fungal biological activities against six phytopathogens.

    Science.gov (United States)

    Huang, Liang-Zhu; Han, Pan; Li, You-Qiang; Xu, Ying-Meng; Zhang, Tao; Du, Zhen-Ting

    2013-09-12

    A series of diaryl amines, ethers and thioethers were synthesized under microwave irradiation efficiently at presence of KF/Al2O3 in 83%-96% yields without any solvent. The salient characters of this method lie in short reaction time, high yields, general applicability to substrates and simple workup procedure. At the same time, their antifungal biological activities against six phytopathogen were evaluated. Most of the compounds (3b, 3c, 3g-o) are more potent than thiophannate-methyl against to Magnaporthe oryzae. This implies that diaryl amine or ether moiety may be helpful in finding a fungicide against Magnaporthe oryzae.

  20. A Facile and Efficient Synthesis of Diaryl Amines or Ethers under Microwave Irradiation at Presence of KF/Al2O3 without Solvent and Their Anti-Fungal Biological Activities against Six Phytopathogens

    Directory of Open Access Journals (Sweden)

    Tao Zhang

    2013-09-01

    Full Text Available A series of diaryl amines, ethers and thioethers were synthesized under microwave irradiation efficiently at presence of KF/Al2O3 in 83%–96% yields without any solvent. The salient characters of this method lie in short reaction time, high yields, general applicability to substrates and simple workup procedure. At the same time, their antifungal biological activities against six phytopathogen were evaluated. Most of the compounds (3b, 3c, 3g–o are more potent than thiophannate-methyl against to Magnaporthe oryzae. This implies that diaryl amine or ether moiety may be helpful in finding a fungicide against Magnaporthe oryzae.

  1. CHARACTERIZATION OF PHYTOPATHOGENIC FUNGI, BACTERIA, NEMATODES AND VIRUSES IN FOUR COMMERCIAL VARIETIES OF HELICONIA (Heliconia sp. CARACTERIZACIÓN DE HONGOS, BACTERIAS, NEMÁTODOS Y VIRUS FITOPATÓGENOS EN CUATRO VARIEDADES COMERCIALES DE HELICONIA (Heliconia sp.

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    Nathali López Cardona

    2012-06-01

    Full Text Available Abstract. Analysis of 914 samples of roots, rhizomes, pseudostems, inflorescences and leaves of four commercial varieties of heliconia, cultivated at the municipality of Chinchiná-Caldas (Colombia, allowed to identify five genera of plant pathogenic fungi (Rhizoctonia, Fusarium, Colletotrichum, Helminthosporium and Curvularia, three genera of plant pathogenic bacteria (Ralstonia, Pseudomonas and Erwinia, two species of viruses (Banana streak virus (BSV, Badnavirus, and Cucumber mosaic virus (CMV, Cucumovirus,, and seven genera of plant parasitic nematodes (Helicotylenchus, Tylenchus, Meloidogyne, Ditylenchus, Aphelenchoides, Pratylenchus, and Radopholus. Of these, Fusarium sp., affecting pseudostems, Pseudomonas sp., affecting leaves and inflorescences, and the plant parasitic nematodes Ditylenchus sp., Aphelenchoides sp., Pratylenchus sp. and Radopholus sp., are new records in the heliconia production in Colombia . The most limiting diseases corresponded to leaf blight, caused by Helminthosporium sp.; the bacterioses, caused by Pseudomonas sp.; the spotted stems, caused by Fusarium sp.; and soft rot of the pseudostems, caused by Erwinia sp. The pathogenicity tests demonstrated that Colletotrichum sp. and Phoma sp. are not pathogenic in leaves; while Fusarium sp., inoculated in pseudostems, Helminthosporium sp. and Pseudomonas sp., inoculated in leaves, and Colletotrichum sp. and Pseudomonas sp., inoculated in inflorescences, had incidence values of 83.3, 86.6, 93.3, 100.0 and 100.0%, respectively.Resumen. El análisis de 914 muestras de raíces, rizomas, pseudotallos, inflorescencias y hojas de cuatro variedades comerciales de heliconia, cultivadas en el municipio de Chinchiná-Caldas (Colombia, permitieron identificar cinco géneros de hongos fitopatógenos (Rhizoctonia, Fusarium, Colletotrichum, Helminthosporium y Curvularia, tres géneros de bacterias fitopatógenas (Ralstonia, Pseudomonas y Erwinia, dos especies de virus (Banana streak

  2. Antibiosis and dark-pigments secretion by the phytopathogenic and environmental fungal species after interaction in vitro with a Bacillus subtilis isolate

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    Alexandre Paulo Machado

    2010-10-01

    Full Text Available In this work, different reactions in vitro between an environmental bacterial isolate and fungal species were related. The Gram-positive bacteria had terminal and subterminal endospores, presented metabolic characteristics of mesophilic and acidophilic growth, halotolerance, positive to nitrate reduction and enzyme production, as caseinase and catalase. The analysis of partial sequences containing 400 to 700 bases of the 16S ribosomal RNA gene showed identity with the genus Bacillus. However, its identity as B. subtilis was confirmed after analyses of the rpoB, gyrA, and 16S rRNA near-full-length sequences. Strong inhibitory activity of environmental microorganisms, such as Penicillium sp, Aspergillus flavus, A. niger, and phytopathogens, such as Colletotrichum sp, Alternaria alternata, Fusarium solani and F. oxysporum f.sp vasinfectum, was shown on co-cultures with B. subtilis strain, particularly on Sabouraud dextrose agar (SDA and DNase media. Red and red-ochre color pigments, probably phaeomelanins, were secreted by A. alternata and A. niger respectively after seven days of co-culture.Na presente investigação, nosso objetivo principal foi relatar diferentes interações in vitro de um isolado bacteriano ambiental com espécies fúngicas. Através da identificação clássica, nós verificamos que o bacilo ambiental apresentava endósporos terminais e subterminais, características metabólicas de mesofilia, acidofilia, halotolerância, redução de nitrato e produção de enzimas, como caseinase e catalase. Análise de seqüências parciais do gene 16S RNAr contendo de 400 a 700 bases revelou identidade com gênero Bacillus. No entanto, a espécie Bacillus subtilis foi confirmada somente depois da análise de seqüências dos genes rpoB, gyrA, and 16S RNAr. Intensa atividade inibitória aos fungos ambientais, como Penicillium sp, Aspergillus flavus, A. niger, e fitopatogênicos, como Colletotrichum sp, Alternaria alternata, Fusarium solani

  3. Inactivation of rsmA leads to overproduction of extracellular pectinases, cellulases, and proteases in Erwinia carotovora subsp. carotovora in the absence of the starvation/cell density-sensing signal, N-(3-oxohexanoyl)-L-homoserine lactone.

    Science.gov (United States)

    Chatterjee, A; Cui, Y; Liu, Y; Dumenyo, C K; Chatterjee, A K

    1995-05-01

    The soft-rotting bacterium, Erwinia carotovora subsp. carotovora 71, produces extracellular enzymes such as pectate lyase isozymes (Pels), cellulase (Cel), polygalacturonase (Peh), and protease (Prt). While the extracellular levels of these enzymes are extremely low when the bacterium is grown in salts-yeast extract-glycerol (SYG) medium, the enzymatic activities are highly induced in SYG medium supplemented with celery extract. By transposon (mini-Tn5) mutagenesis, we isolated a RsmA- mutant, AC5070, which overproduces extracellular enzymes; the basal levels of Pel, Peh, and Cel in AC5070 are higher than the induced levels in the RsmA+ parent, AC5047. While Peh production is mostly constitutive in AC5070, Pel, Cel, and Prt production is still inducible with celery extract. The high basal levels of pel-1, pel-3, and peh-1 mRNAs in AC5070 demonstrate that overproduction of the pectolytic enzymes is due to the stimulation of transcription. Using chromosomal DNA flanking mini-Tn5 as a probe, we cloned the wild-type rsmA+ allele, which suppresses Pel, Peh, Cel, and Prt production in both RsmA+ and RsmA- strains. The RsmA- mutant, like its parent, produces N-(3-oxohexanoyl)-L-homoserine lactone (HSL), a starvation/cell density-sensing signal required for extracellular enzyme production. To examine the role of HSL, we constructed HSL-deficient strains by replacing hslI, a locus required for HSL production, with hslI::Tn3HoHo1-Spc. While the basal levels of Pel, Peh, Cel, and Prt are comparable in the RsmA- mutant and its HSL- derivative, these enzymes are barely detectable in the Hsl- derivative of the RsmA+ parent strain.(ABSTRACT TRUNCATED AT 250 WORDS)

  4. 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. Copyright © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  5. Genetic analysis of streptomycin-resistant (Sm(R)) strains of Erwinia amylovora suggests that dissemination of two genotypes is responsible for the current distribution of Sm(R) E. amylovora in Michigan.

    Science.gov (United States)

    McGhee, Gayle C; Guasco, Jesse; Bellomo, Lisa M; Blumer-Schuette, Sara E; Shane, William W; Irish-Brown, Amy; Sundin, George W

    2011-02-01

    Streptomycin-resistant (Sm(R)) strains of the fire blight pathogen Erwinia amylovora were first isolated in southwest Michigan in 1991. Since that time, resistant strains have progressed northward to other apple-producing regions in the state. A total of 98.7% of Sm(R) strains isolated between 2003 and 2009 in Michigan harbored the strA-strB genes on transposon Tn5393. strA and strB encode phosphotransferase enzymes that modify streptomycin to a nonbactericidal form. Mutational resistance to streptomycin, caused by a point mutation-mediated target-site alteration of the ribosomal S12 protein, occurred in 1.3% of E. amylovora strains from Michigan. Tn5393 was originally introduced to E. amylovora on the plasmid pEa34; thus, the first Sm(R) strains isolated contained both pEa34 and the ubiquitous nonconjugative plasmid pEA29. More recently, we have observed Sm(R) strains in which Tn5393 is present on pEA29, suggesting that the transposon has moved via transposition from pEa34 to pEA29. Almost all of the strains containing Tn5393 on pEA29 had lost pEa34. Of 210 pEA29::Tn5393 plasmids examined, the transposon was inserted at either nucleotide position 1,515 or 17,527. Both of these positions were in noncoding regions of pEA29. Comparative sequencing of the housekeeping genes groEL and potentially variable sequences on pEA29 was done in an attempt to genetically distinguish Sm(R) strains from streptomycin-sensitive (Sm(S)) strains isolated in Michigan. Only 1 nucleotide difference within the total 2,660 bp sequenced from each strain was observed in 2 of 29 strains; multiple sequence differences were observed between the Michigan strains and E. amylovora control strains isolated in the western United States or from Rubus spp. Alterations in virulence observable using an immature pear fruit assay were detected in three of eight Sm(R) strains examined. Our current genetic data indicate that only two Sm(R) strain genotypes (strains containing pEA29::Tn5393 with Tn5393

  6. Erwinia resistentie in Zantedeschia : eindverslag juni 2003

    NARCIS (Netherlands)

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

    2003-01-01

    Het gewas Zantedeschia heeft zich de afgelopen jaren razendsnel ontwikkeld als nieuw siergewas. De veredeling van Zantedeschia staat echter nog in de kinderschoenen. De ontwikkeling van kennis, technieken en materiaal op het gebied van Zantedeschia is van belang met het oog op de positie die het

  7. Antifungal activity of medicinal plant extracts against phytopathogenic fungus Alternaria spp Actividad antifúngica de extractos de plantas medicinales contra el hongo fitopatógeno Alternaria spp

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    Paola Díaz Dellavalle

    2011-06-01

    Full Text Available The aim of the study was to evaluate the antifungal activity of extracts of 10 plant species used in traditional Uruguayan medicine against the phytopathogenic fungus Alternaria spp. The plants were selected on the basis of their reported ethnobotanical uses. Aqueous, saline buffer and acid extracts of different plant species were screened in vitro for their antifungal activity against Alternaria spp. For the antifungal evaluation we used a microspectrophotometric assay. Minimal inhibitory concentration (MIC and minimum fungicidal concentration (MFC of the extracts were determined. Three solvents were assayed on different tissues of the plants and among the 29 evaluated extracts, 31% of the extracts inhibited growth, similar to the effects of a chemical fungicide. Acid extracts of the plants were more effective than the aqueous or buffer extracts against Alternaria spp. The MIC values of the extracts were determined ranging between 1.25 and 25 µg mL-1. The MFC values of the extracts ranged between 1.25 µg mL-1 (Rosmarinus officinalis L. and 10 µg mL-1 (Cynara scolymus L.. MICs and MFCs values obtained from leaves (Salvia officinalis L. and R. officinalis and seeds extracts (Salvia sclarea L. were quite comparable to values obtained with the conventional fungicide captan (2.5 µg mL-1. The extracts of Salvia sclarea, S. officinalis and R. officinalis could be considered as potential sources of antifungal compounds for treating diseases in plants. These extracts showed maximum activity, even at very low concentrations, and the same fungicide effects as chemical fungicide. We conclude from this that these extracts exhibit amazing fungicidal properties that support their traditional use as antiseptics.El objetivo de este trabajo fue evaluar la actividad antifúngica de extractos vegetales de 10 especies utilizadas en la medicina tradicional uruguaya contra el hongo fitopatógeno Alternaria spp. Las plantas fueron seleccionadas en base a usos

  8. Identificarea izolatului bacterian P5 obţinut din plante de măr

    OpenAIRE

    MAGHER Maria

    2016-01-01

    The phytopathogen Erwinia amylovora (Burrill) Winslow et al. is a bacterium having a major impact on pome fruit species causing identification problems at some development phenophases of trees. The aim of this paper was to identify the bacterial isolate P5 obtained from apple tree samples using classical and modern methods recommended for the bacterium E. amylovora (API 20E test, gas chromatography, PCR, DASELISA and indirect immunofluorescence). As a result of our investigations, the bacteri...

  9. Changes in transcriptional orientation are associated with increases in evolutionary rates of enterobacterial genes

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    Hsiung Chao

    2011-10-01

    Full Text Available Abstract Background Changes in transcriptional orientation (“CTOs” occur frequently in prokaryotic genomes. Such changes usually result from genomic inversions, which may cause a conflict between the directions of replication and transcription and an increase in mutation rate. However, CTOs do not always lead to the replication-transcription confrontation. Furthermore, CTOs may cause deleterious disruptions of operon structure and/or gene regulations. The currently existing CTOs may indicate relaxation of selection pressure. Therefore, it is of interest to investigate whether CTOs have an independent effect on the evolutionary rates of the affected genes, and whether these genes are subject to any type of selection pressure in prokaryotes. Methods Three closely related enterbacteria, Escherichia coli, Klebsiella pneumoniae and Salmonella enterica serovar Typhimurium, were selected for comparisons of synonymous (dS and nonsynonymous (dN substitution rate between the genes that have experienced changes in transcriptional orientation (changed-orientation genes, “COGs” and those that do not (same-orientation genes, “SOGs”. The dN/dS ratio was also derived to evaluate the selection pressure on the analyzed genes. Confounding factors in the estimation of evolutionary rates, such as gene essentiality, gene expression level, replication-transcription confrontation, and decreased dS at gene terminals were controlled in the COG-SOG comparisons. Results We demonstrate that COGs have significantly higher dN and dS than SOGs when a series of confounding factors are controlled. However, the dN/dS ratios are similar between the two gene groups, suggesting that the increase in dS can sufficiently explain the increase in dN in COGs. Therefore, the increases in evolutionary rates in COGs may be mainly mutation-driven. Conclusions Here we show that CTOs can increase the evolutionary rates of the affected genes. This effect is independent of the replication-transcription confrontation, which is suggested to be the major cause of inversion-associated evolutionary rate increases. The real cause of such evolutionary rate increases remains unclear but is worth further explorations.

  10. Biophysical analysis of the interaction of granulysin-derived peptides with enterobacterial endotoxins.

    Science.gov (United States)

    Chen, Xi; Howe, Jörg; Andrä, Jörg; Rössle, Manfred; Richter, Walter; da Silva, Ana Paula Galvão; Krensky, Alan M; Clayberger, Carol; Brandenburg, Klaus

    2007-10-01

    To combat infections by Gram-negative bacteria, it is not only necessary to kill the bacteria but also to neutralize pathogenicity factors such as endotoxin (lipopolysaccharide, LPS). The development of antimicrobial peptides based on mammalian endotoxin-binding proteins is a promising tool in the fight against bacterial infections, and septic shock syndrome. Here, synthetic peptides derived from granulysin (Gra-pep) were investigated in microbiological and biophysical assays to understand their interaction with LPS. We analyzed the influence of the binding of Gra-pep on (1) the acyl chain melting of the hydrophobic moiety of LPS, lipid A, by Fourier-transform spectroscopy, (2) the aggregate structure of LPS by small-angle X-ray scattering and cryo-transmission electron microscopy, and 3) the enthalpy change by isothermal titration calorimetry. In addition, the influence of Gra-pep on the incorporation of LPS and LPS-LBP (lipopolysaccharide-binding protein) complexes into negatively charged liposomes was monitored. Our findings demonstrate a characteristic change in the aggregate structure of LPS into multilamellar stacks in the presence of Gra-pep, but little or no change of acyl chain fluidity. Neutralization of LPS by Gra-pep is not due to a scavenging effect in solution, but rather proceeds after incorporation into target membranes, suggesting a requisite membrane-bound step.

  11. In vitro synthesis of intermediates involved in the assembly of enterobacterial common antigen (ECA)

    International Nuclear Information System (INIS)

    Barr, K.; Wolski, S.; Kroto, J.; Rick, P.D.

    1986-01-01

    ECA is a cell surface antigen found in all bacteria belonging to the family Enterobacteriaceae. The serological specificity of ECA is determined by a linear heteropolysaccharide comprised of trisaccharide repeat units; the component sugars are N-acetyl-D-glucosamine (GlcNAc), N-acetyl-D-mannosaminuronic acid (ManNAcUA), and 4-acetamido-D-fucose (Fuc4NAc). In vivo studies have suggested that GlcNAc-pyrophosphorylundecaprenol (GlcNAc-PP-lipid) is an intermediate in ECA synthesis. More recently, they have demonstrated UDP-GlcNAc:undecaprenylphosphate GlcNAc-1-phosphate transferase activity in cell envelope preparations of E. coli. Radioactivity from UDP-[ 3 H]Glc-NAc was incorporated into endogenous lipid acceptor, and the labeled product was characterized as GlcNAc-PP-lipid (lipid I). Transferase activity was inhibited by tunicamycin and UMP, but it was unaffected by UDP. The reaction was reversible, and the synthesis of UDP-[ 3 H]GlcNAc from UMP and [ 3 H]GlcNAc-PP-lipid was also sensitive to tunicamycin. The simultaneous addition of UDP-[ 14 C]ManNAcUA and UDP-[ 3 H]GlcNAc to cell envelope preparations resulted in the synthesis of a more polar lipid (lipid II) that contained both labeled sugars in equimolar amounts. Synthesis of lipid II was dependent on prior synthesis of lipid I. Accordingly, [ 3 H]GlcNAc-PP-lipid that had been synthesized in vivo served as an acceptor in vitro of ManNAcUA residues from UDP-ManNAcUA. Lipid II has been tentatively identified as ManNAcUA-GlcNAc-pyrophosphorylundecaprenol

  12. Microarray comparative genomic hybridisation analysis incorporating genomic organisation, and application to enterobacterial plant pathogens.

    Directory of Open Access Journals (Sweden)

    Leighton Pritchard

    2009-08-01

    Full Text Available Microarray comparative genomic hybridisation (aCGH provides an estimate of the relative abundance of genomic DNA (gDNA taken from comparator and reference organisms by hybridisation to a microarray containing probes that represent sequences from the reference organism. The experimental method is used in a number of biological applications, including the detection of human chromosomal aberrations, and in comparative genomic analysis of bacterial strains, but optimisation of the analysis is desirable in each problem domain.We present a method for analysis of bacterial aCGH data that encodes spatial information from the reference genome in a hidden Markov model. This technique is the first such method to be validated in comparisons of sequenced bacteria that diverge at the strain and at the genus level: Pectobacterium atrosepticum SCRI1043 (Pba1043 and Dickeya dadantii 3937 (Dda3937; and Lactococcus lactis subsp. lactis IL1403 and L. lactis subsp. cremoris MG1363. In all cases our method is found to outperform common and widely used aCGH analysis methods that do not incorporate spatial information. This analysis is applied to comparisons between commercially important plant pathogenic soft-rotting enterobacteria (SRE Pba1043, P. atrosepticum SCRI1039, P. carotovorum 193, and Dda3937.Our analysis indicates that it should not be assumed that hybridisation strength is a reliable proxy for sequence identity in aCGH experiments, and robustly extends the applicability of aCGH to bacterial comparisons at the genus level. Our results in the SRE further provide evidence for a dynamic, plastic 'accessory' genome, revealing major genomic islands encoding gene products that provide insight into, and may play a direct role in determining, variation amongst the SRE in terms of their environmental survival, host range and aetiology, such as phytotoxin synthesis, multidrug resistance, and nitrogen fixation.

  13. Ação do óleo essencial de Syzygium aromaticum (L. Merr. & L.M.Perry sobre as hifas de alguns fungos fitopatogênicos Action of Syzygium aromaticum (L. Merr. & L.M.Perry essential oil on the hyphae of some phytopathogenic fungi

    Directory of Open Access Journals (Sweden)

    A.R.T Costa

    2011-01-01

    Full Text Available Atualmente o uso de métodos alternativos para o controle de doenças e pragas na agricultura, visando minimizar os danos ao meio ambiente e à saúde pública é uma prática reconhecida e necessária. Este trabalho objetivou investigar a ação do óleo essencial de Syzygium aromaticum (L. Merr. & L.M.Perry sobre o crescimento micelial in vitro dos fungos fitopatogênicos Rhizoctonia solani, Fusarium solani, Fusarium oxysporum e Macrophomina phaseolina. A análise por cromatografia gasosa acoplada com espectrometria de massa possibilitou a identificação de eugenol (83,6%, acetato de eugenila (11,6% e cariofileno (4,2%. A avaliação microscópica dos micélios dos fungos evidenciou diversas alterações morfológicas, como a presença de vacúolos, desorganização dos conteúdos celulares, diminuição na nitidez da parede celular, intensa fragmentação e menor turgência das hifas. O óleo essencial de cravo apresentou atividade fungicida na concentração de 0,15% sobre o crescimento de R. solani, F. oxysporum e F. solani, entretanto não demonstrou essa atividade sobre M. phaseolina. Esses resultados indicam perspectivas favoráveis para posterior uso do óleo de cravo no controle desses fitopatógenos na agricultura.Currently, the use of alternative methods to control diseases and pests in agriculture has been a recognized and necessary practice to minimize damages to the environment and public health. This study aimed to investigate the action of clove [Syzygium aromaticum (L. Merr. & L.M.Perry] essential oil on the in vitro mycelial growth of the phytopathogenic fungi Rhizoctonia solani, Fusarium solani, Fusarium oxysporum and Macrophomina phaseolina. Analysis by gas chromatography-mass spectrometry allowed the identification of eugenol (83.6%, eugenyl acetate (11.6% and caryophyllene (4.2%. Microscopic evaluation of mycelia showed several morphological changes such as presence of vacuoles, cell content disorganization, decreased

  14. Membrane Vesicles and Lactamase in Erwinia herbicola Essam A ...

    African Journals Online (AJOL)

    yakoub@AHMED

    ... parent strain (Kadurugamuwa et al. 1998). Beveridge (1999) reported that MVs are spherical bilayered membranous structures (50 - 250 nm in diameter) released from the surface of gram-negative bacteria. Hayashi et al. (2002) concluded that an autolysin mutant of Porphyromonas gingivalis constructed and produced ...

  15. Erwinia pyrifoliae, a new pathogen on strawberry in the Netherlands

    NARCIS (Netherlands)

    Wenneker, M.; Bergsma-Vlami, M.M.

    2015-01-01

    BACKGROUND: During the late spring of 2013 strawberry plants grown under protection (Fragaria x ananassa cv. Elsanta) were found at several locations in the Netherlands showing an intense brown to black discoloration of their immature fruits, their fruit calyx and the attached stems. OBJECTIVE:

  16. The Structural Basis of Erwinia rhapontici Isomaltulose Synthase

    Science.gov (United States)

    Xu, Zheng; Li, Sha; Li, Jie; Li, Yan; Feng, Xiaohai; Wang, Renxiao; Xu, Hong; Zhou, Jiahai

    2013-01-01

    Sucrose isomerase NX-5 from Erwiniarhapontici efficiently catalyzes the isomerization of sucrose to isomaltulose (main product) and trehalulose (by-product). To investigate the molecular mechanism controlling sucrose isomer formation, we determined the crystal structures of native NX-5 and its mutant complexes E295Q/sucrose and D241A/glucose at 1.70 Å, 1.70 Å and 2.00 Å, respectively. The overall structure and active site architecture of NX-5 resemble those of other reported sucrose isomerases. Strikingly, the substrate binding mode of NX-5 is also similar to that of trehalulose synthase from Pseudomonasmesoacidophila MX-45 (MutB). Detailed structural analysis revealed the catalytic RXDRX motif and the adjacent 10-residue loop of NX-5 and isomaltulose synthase PalI from Klebsiella sp. LX3 adopt a distinct orientation from those of trehalulose synthases. Mutations of the loop region of NX-5 resulted in significant changes of the product ratio between isomaltulose and trehalulose. The molecular dynamics simulation data supported the product specificity of NX-5 towards isomaltulose and the role of the loop330-339 in NX-5 catalysis. This work should prove useful for the engineering of sucrose isomerase for industrial carbohydrate biotransformations. PMID:24069347

  17. In vitro effect of Bacillus thuringiensis strains and Cry proteins in phytopathogenic fungi of paddy rice-field Efeito in vitro de cepas e proteínas Cry de Bacillus thuringiensis em fungos fitopatogênicos da cultura do arroz irrigado

    Directory of Open Access Journals (Sweden)

    Neiva Knaak

    2007-09-01

    Full Text Available Cry1Ab and Cry1Ac strains and proteins synthesized by Bacillus thuringiensis thuringiensis and B. thuringiensis kurstaki were assessed in the following phytopathogens: Rhizoctonia solani,Pyricularia grisea,Fusarium oxysporum and F. solani, which had their micelial growth decreased after incubation in the presence of the bacterial strains. As to Cry proteins, there were no inhibition halo development in the assessed concentrations.As cepas e proteínas Cry1Ab e Cry1Ac sintetizadas por Bacillus thuringiensis thuringiensis e B. thuringiensis kurstaki, foram avaliadas nos fitopatógenos: Rhizoctonia solani,Pyricularia grisea,Fusarium oxysporum e F. solani, os quais tiveram seu crescimento micelial reduzido após a incubação na presença das cepas bacterianas. Em relação às proteínas Cry, não houve formação de halo de inibição nas concentrações avaliadas.

  18. Selection of endophytic fungi from comfrey (Symphytum officinale L. for in vitro biological control of the phytopathogen Sclerotinia sclerotiorum (Lib. Seleção de fungos endofíticos de confrei (Symphytum officinale L., buscando controle biológico in vitro do fitopatógeno Sclerotinia sclerotiorum (Lib.

    Directory of Open Access Journals (Sweden)

    Rafaeli Rocha

    2009-03-01

    Full Text Available Biological control consists of using one organism to attack another that may cause economic damage to crops. Integrated Pest Management (IPM is a very common strategy. The white mold produced by Sclerotiniasclerotiorum (Lib. causes considerable damage to bean crops. This fungus is a soil inhabitant, the symptoms of which are characterized by water-soaked lesions covered by a white cottony fungal growth on the soil surface and/or the host plant. Possible biological control agents taken from plants are being investigated as phytopathogen inhibitors. These are endophytic microorganisms that inhabit the intercellular spaces of vegetal tissues and are often responsible for antimicrobial production. The objective of the present study was to select endophytic fungi isolated from comfrey (Symphytumofficinale L. leaves with in vitro antagonist potential against the phytopathogenic fungus S. sclerotiorum. Twelve isolates of endophytic fungi and a pathogenic strain of S. sclerotiorum were used in the challenge method. With the aid of this method, four endophytes with the best antagonistic activity against S. sclerotiorum were selected. Pathogen growth inhibition zones were considered indicative of antibiosis. The percentages of pathogenic mycelia growth were measured both with and without the antagonist, resulting in growth reductions of 46.7% to 50.0% for S. sclerotiorum. These analyses were performed by evaluating the endophytic/pathogenic mycelia growth in mm/day over an eight-day period of antagonistic tests.O controle biológico consiste no uso de organismos que atacam outros que causam danos a culturas de plantas. Esta é uma estratégia muito utilizada no Controle Integrado de Pragas (CIP. O mofo branco, causado por Sclerotiniasclerotiorum (Lib., causa danos em culturas de feijão. Este fungo é encontrado no solo e seus sintomas são caracterizados por lesões úmidas cobertas por micélios algodonosos, crescidos a partir do solo e/ou da planta

  19. Atividade antimicrobiana de óleos essenciais no controle de alguns fitopatógenos fúngicos in vitro e no tratamento de sementes Antimicrobial activity of essential oils on the in vitro control of some fungal phytopathogens and on seed treatment

    Directory of Open Access Journals (Sweden)

    T. Hillen

    2012-01-01

    Full Text Available Este trabalho verificou o efeito dos óleos essenciais (OE extraídos de Eremanthus erythropappus (candeia, Cymbopogon martinii (palmarosa e de Rosmarinus officinalis (alecrim no crescimento micelial de alguns fitopatógenos fúngicos e no tratamento de sementes de milho, soja e feijão. No teste in vitro, alíquotas de 20, 40, 60, 100, 200, 500 e 1000 μL de cada um dos óleos essenciais foram distribuídas na superfície do meio de cultura. Posteriormente, discos de meio de cultura com micélio de Alternaria carthami, Alternaria sp. e Rhizoctonia solani foram transferidos para o centro de cada placa. O crescimento foi mensurado e calculada a taxa de inibição do crescimento micelial (ICM. Para verificar o efeito dos OE na germinação das sementes utilizou-se a aplicação deles por fumigação. Foi avaliada a percentagem de sementes germinadas e a incidência de patógenos nas sementes. Sobre o crescimento micelial, o óleo de palmarosa inibiu completamente todos os patógenos fúngicos, independentemente da concentração. Já os óleos de candeia e alecrim foram melhores quando foram adicionadas alíquotas superiores a 200 μL. Os óleos influenciaram diferentemente a germinação e a sanidade das sementes de milho, soja e feijão.This study aimed to verify the effect of essential oils (EO extracted from Eremanthus erythropappus ("candeia" Cymbopogon martinii ("palmarosa" and Rosmarinus officinalis (rosemary on the mycelial growth of some fungal phytopathogens, as well as on the treatment of corn, soybean and bean seeds. In the in vitro test, aliquots of 20, 40, 60, 100, 200, 500 and 1000 μL of each essential oil were distributed on the surface of the culture medium. Then, discs of culture medium with mycelium of Alternaria carthami, Alternaria sp and Rhizoctonia solani were transferred to the center of each plate. Growth was measured and the mycelial growth inhibition rate (MGI was calculated. To verify the effect of EO on seed

  20. Identificarea izolatului bacterian P5 obţinut din plante de măr

    Directory of Open Access Journals (Sweden)

    MAGHER Maria

    2016-06-01

    Full Text Available The phytopathogen Erwinia amylovora (Burrill Winslow et al. is a bacterium having a major impact on pome fruit species causing identification problems at some development phenophases of trees. The aim of this paper was to identify the bacterial isolate P5 obtained from apple tree samples using classical and modern methods recommended for the bacterium E. amylovora (API 20E test, gas chromatography, PCR, DASELISA and indirect immunofluorescence. As a result of our investigations, the bacterium Erwinia amylovora was clearly identified. Taking into consideration the quarantine status of the phytopathogen E. amylovora, it is necessary to carry out the phytosanitary test sin order to monitor the presence of fire blight on pome fruit species as well as to implement timely the required preventive and curative control measures. Rezumat. Fitopatogenul Erwinia amylovora (Burrill Winslow et al. este o bacterie cu un impact deosebit asupra speciilor pomicole sămânţoase, care prezintă probleme de identificare la anumite fenofaze de dezvoltare a pomilor. Scopul acestei lucrări a fost de a identifica izolatul bacterian P5, obţinut din probe de măr, prin metode clasice şi contemporane recomandate pentru bacteria E. amylovora (testul API 20E, gaz-cromatografie, PCR, DAS-ELISA, imunofluorescenţă indirectă. În rezultatul cercetărilor a fost identificată bacteria Erwinia amylovora. Având în vedere statutul de carantină al fitopatogenului E. amylovora, se impune necesitatea efectuării obligatorii a sondajelor fitosanitare pentru monitorizarea prezenţei focului bacterian la culturile pomicole sămânţoase şi efectuarea la timp a măsurilor preventive şi curative de combatere.

  1. Tolerogenic dendritic cells pulsed with enterobacterial extract suppress development of colitis in the severe combined immunodeficiency transfer model

    DEFF Research Database (Denmark)

    Pedersen, Anders Elm; Gad, M; Kristensen, N N

    2007-01-01

    and experimental allergic encephalitis, but the effect in inflammatory bowel disease has not yet been demonstrated. In severe combined immunodeficient (SCID) mice, adoptively transferred CD4(+) CD25(-) T cells repopulate the lymphoid tissues and lead to development of chronic colitis characterized by CD4(+) T...

  2. Myconanoparticles: synthesis and their role in phytopathogens management

    Science.gov (United States)

    Alghuthaymi, Mousa A.; Almoammar, Hassan; Rai, Mahindra; Said-Galiev, Ernest; Abd-Elsalam, Kamel A.

    2015-01-01

    Nanotechnology can offer green and eco-friendly alternatives for plant disease management. Apart from being eco-friendly, fungi are used as bio-manufacturing units, which will provide an added benefit in being easy to use, as compared to other microbes. The non-pathogenic nature of some fungal species in combination with the simplicity of production and handling will improve the mass production of silver nanoparticles. Recently, a diverse range of fungi have been screened for their ability to create silver nanoparticles. Mycosynthesis of gold, silver, gold–silver alloy, selenium, tellurium, platinum, palladium, silica, titania, zirconia, quantum dots, usnic acid, magnetite, cadmium telluride and uraninite nanoparticles has also been reported by various researchers. Nanotechnological application in plant pathology is still in the early stages. For example, nanofungicides, nanopesticides and nanoherbicides are being used extensively in agriculture practices. Remote activation and monitoring of intelligent nano-delivery systems can assist agricultural growers of the future to minimize fungicides and pesticides use. Nanoparticle-mediated gene transfer would be useful for improvement of crops resistant to pathogens and pest. This review critically assesses the role of fungi in the synthesis of nanoparticles, the mechanism involved in the synthesis, the effect of different factors on the reduction of metal ions in developing low-cost techniques for the synthesis and recovery of nanoparticles. Moreover, the application of nanoparticles in plant disease control, antimicrobial mechanisms, and nanotoxicity on plant ecosystem and soil microbial communities has also been discussed in detail. PMID:26019636

  3. Inhibition of growth of some phytopathogenic and mycotoxigenic ...

    African Journals Online (AJOL)

    STORAGESEVER

    2008-08-18

    Aug 18, 2008 ... usage have been undertaken to reduce or prevent conta- mination of ... Fungicide usage is more feasible but the effects of fungicides on non-target organisms coupled with their poor biodegradability are unwelcome. Other ways of ... ted in a plastic bag and transported to the laboratory at the. University of ...

  4. Antifungal activity of rice straw extract on some phytopathogenic fungi

    African Journals Online (AJOL)

    user

    2012-09-04

    Sep 4, 2012 ... Key words: Rice straw, allelochemicals, antifungal, Aspergillus flavus, Alternaria alternata, Botrytis cinerea, amylase, protease ..... Identification and quantification of compounds in a series of allelopathic and non- allelopathic rice root exudates. J. Chem. Ecol. 30:1647-1662. Timmer LW, Peever TL, Solel Z, ...

  5. Reactive oxygen species in phytopathogenic fungi: signaling, development, and disease.

    Science.gov (United States)

    Heller, Jens; Tudzynski, Paul

    2011-01-01

    Reactive oxygen species (ROS) play a major role in pathogen-plant interactions: recognition of a pathogen by the plant rapidly triggers the oxidative burst, which is necessary for further defense reactions. The specific role of ROS in pathogen defense is still unclear. Studies on the pathogen so far have focused on the importance of the oxidative stress response (OSR) systems to overcome the oxidative burst or of its avoidance by effectors. This review focuses on the role of ROS for fungal virulence and development. In the recent years, it has become obvious that (a) fungal OSR systems might not have the predicted crucial role in pathogenicity, (b) fungal pathogens, especially necrotrophs, can actively contribute to the ROS level in planta and even take advantage of the host's response, (c) fungi possess superoxide-generating NADPH oxidases similar to mammalian Nox complexes that are important for pathogenicity; however, recent data indicate that they are not directly involved in pathogen-host communication but in fungal differentiation processes that are necessary for virulence. Copyright © 2011 by Annual Reviews. All rights reserved.

  6. Anti-phytopathogenic activities of macro-algae extracts.

    Science.gov (United States)

    Jiménez, Edra; Dorta, Fernando; Medina, Cristian; Ramírez, Alberto; Ramírez, Ingrid; Peña-Cortés, Hugo

    2011-01-01

    Aqueous and ethanolic extracts obtained from nine Chilean marine macro-algae collected at different seasons were examined in vitro and in vivo for properties that reduce the growth of plant pathogens or decrease the injury severity of plant foliar tissues following pathogen infection. Particular crude aqueous or organic extracts showed effects on the growth of pathogenic bacteria whereas others displayed important effects against pathogenic fungi or viruses, either by inhibiting fungal mycelia growth or by reducing the disease symptoms in leaves caused by pathogen challenge. Organic extracts obtained from the brown-alga Lessonia trabeculata inhibited bacterial growth and reduced both the number and size of the necrotic lesion in tomato leaves following infection with Botrytis cinerea. Aqueous and ethanolic extracts from the red-alga Gracillaria chilensis prevent the growth of Phytophthora cinnamomi, showing a response which depends on doses and collecting-time. Similarly, aqueous and ethanolic extracts from the brown-alga Durvillaea antarctica were able to diminish the damage caused by tobacco mosaic virus (TMV) in tobacco leaves, and the aqueous procedure is, in addition, more effective and seasonally independent. These results suggest that macro-algae contain compounds with different chemical properties which could be considered for controlling specific plant pathogens.

  7. Anti-Phytopathogenic Activities of Macro-Algae Extracts

    OpenAIRE

    Jiménez, Edra; Dorta, Fernando; Medina, Cristian; Ramírez, Alberto; Ramírez, Ingrid; Peña-Cortés, Hugo

    2011-01-01

    Aqueous and ethanolic extracts obtained from nine Chilean marine macro-algae collected at different seasons were examined in vitro and in vivo for properties that reduce the growth of plant pathogens or decrease the injury severity of plant foliar tissues following pathogen infection. Particular crude aqueous or organic extracts showed effects on the growth of pathogenic bacteria whereas others displayed important effects against pathogenic fungi or viruses, either by inhibiting fungal myceli...

  8. Inhibition of growth of some phytopathogenic and mycotoxigenic ...

    African Journals Online (AJOL)

    Investigation of the chemistry of the ash using Fourier-Transform Infrared (FT-IR) spectroscopy and Xray powder diffraction (XRPD) analysis revealed the presence of aluminium oxide (Al2O3), calcium carbonate (CaCO3), calcium hydroxide (Ca(OH)2), potassium chloride (KCl), potassium sulphate (K2SO4) and cobalt ...

  9. Biotransformation of the phytoalexin camalexin by the phytopathogen Rhizoctonia solani.

    Science.gov (United States)

    Pedras, M S; Khan, A Q

    2000-01-01

    The unusual metabolism of the cruciferous phytoalexin camalexin by virulent and weakly virulent isolates of the root rot fungus Rhizoctonia solani Kuhn is reported. This biotransformation proceeded via 5-hydroxycamalexin, which was further biotransformed into more polar metabolites. Importantly, the metabolites resulting from transformation of camalexin were significantly less toxic to the pathogen than camalexin. Thus, it was concluded that R. solani can detoxify camalexin through oxidation of the indole ring. The chemistry involved in the structure determination of the intermediates of this pathway, their synthesis as well as antifungal activity is described.

  10. Antifungal activity of rice straw extract on some phytopathogenic fungi

    African Journals Online (AJOL)

    The antifungal activity of allelochemicals extracted from rice straw on the radial growth rate and the activity of some hydrolyzing enzymes of Aspergillus flavus, Alternaria alternata and Botrytis cinerea were studied in vitro. Five different concentrations (2, 4, 6, 8 and 10%, w/v) of water, methanol and acetone extracts of rice ...

  11. Anti-Phytopathogenic Activities of Macro-Algae Extracts

    Directory of Open Access Journals (Sweden)

    Ingrid Ramírez

    2011-05-01

    Full Text Available Aqueous and ethanolic extracts obtained from nine Chilean marine macro-algae collected at different seasons were examined in vitro and in vivo for properties that reduce the growth of plant pathogens or decrease the injury severity of plant foliar tissues following pathogen infection. Particular crude aqueous or organic extracts showed effects on the growth of pathogenic bacteria whereas others displayed important effects against pathogenic fungi or viruses, either by inhibiting fungal mycelia growth or by reducing the disease symptoms in leaves caused by pathogen challenge. Organic extracts obtained from the brown-alga Lessonia trabeculata inhibited bacterial growth and reduced both the number and size of the necrotic lesion in tomato leaves following infection with Botrytis cinerea. Aqueous and ethanolic extracts from the red-alga Gracillaria chilensis prevent the growth of Phytophthora cinnamomi, showing a response which depends on doses and collecting-time. Similarly, aqueous and ethanolic extracts from the brown-alga Durvillaea antarctica were able to diminish the damage caused by tobacco mosaic virus (TMV in tobacco leaves, and the aqueous procedure is, in addition, more effective and seasonally independent. These results suggest that macro-algae contain compounds with different chemical properties which could be considered for controlling specific plant pathogens.

  12. ACTIVITY OF NATURAL PRODUCTS AGAINST SOME PHYTOPATHOGENIC FUNGI.

    Science.gov (United States)

    La Torre, A; Caradonia, F; Gianferro, M; Molinu, M G; Battaglia, V

    2014-01-01

    The requirement of environmental protection and food safety is perceived with always major interest by public opinion and it is consistent with European Union legislation on the sustainable use of pesticides (Directive 2009/128/EC). This directive requires member states to promote low pesticide-input, giving priority to non-chemical methods and low risk plant protection products. In order to contribute to the achievement of these objectives antifungal activity of natural substances, characterized by a good toxicological and ecotoxicological profile, was tested. Essential oil of Melaleuca alternifolia, essential oil of Syzygium aromaticum and extract from Mimosa tenuiflora were tested against Alternaria alternata, Botrytis cinerea and Fusarium oxysporum f. sp. lycopersici (races 1 and 2). In vitro tests involved determination of radial growth of the colonies of fungi in the presence of varying concentrations of tested products in agar media and determination of germination percentage in the presence of tested product at various concentrations. The products based on essential oil of M. alternifolia were also tested in vivo on tomato fruits wounded and artificially inoculated with A. alternata or with B. cinerea. The in vitro tests showed the antifungal activity of both essential oils instead the extract from M. tenuiflora exhibited poor antifungal activity and only against A. alternata and B. cinerea. The results on tomato fruits showed inhibition of grey mould and black mould by essential oil of M. alternifolia. The antifungal activity increased with increasing concentrations. In conclusion, the obtained results in the present study showed promising prospects for the utilisation of investigated products to reduce the using of antifungal chemicals and to achieve a more sustainable use of pesticides.

  13. Myconanoparticles: synthesis and their role in phytopathogens management

    International Nuclear Information System (INIS)

    Alghuthaymi, Mousa A.; Almoammar, Hassan; Rai, Mahindra; Said-Galiev, Ernest; Abd-Elsalam, Kamel A.

    2015-01-01

    Nanotechnology can offer green and eco-friendly alternatives for plant disease management. Apart from being eco-friendly, fungi are used as bio-manufacturing units, which will provide an added benefit in being easy to use, as compared to other microbes. The non-pathogenic nature of some fungal species in combination with the simplicity of production and handling will improve the mass production of silver nanoparticles. Recently, a diverse range of fungi have been screened for their ability to create silver nanoparticles. Mycosynthesis of gold, silver, gold-silver alloy, selenium, tellurium, platinum, palladium, silica, titania, zirconia, quantum dots, usnic acid, magnetite, cadmium telluride and uraninite nanoparticles has also been reported by various researchers. Nanotechnological application in plant pathology is still in the early stages. For example, nanofungicides, nanopesticides and nanoherbicides are being used extensively in agriculture practices. Remote activation and monitoring of intelligent nano-delivery systems can assist agricultural growers of the future to minimize fungicides and pesticides use. Nanoparticle-mediated gene transfer would be useful for improvement of crops resistant to pathogens and pest. This review critically assesses the role of fungi in the synthesis of nanoparticles, the mechanism involved in the synthesis, the effect of different factors on the reduction of metal ions in developing low-cost techniques for the synthesis and recovery of nanoparticles. Moreover, the application of nanoparticles in plant disease control, antimicrobial mechanisms, and nanotoxicity on plant ecosystem and soil microbial communities has also been discussed in detail. Keywords: disease management; antimicrobial action; phytotoxicity; nanophytopathology

  14. The complete mitochondrial genome of the phytopathogenic fungus Phomopsis longicolla

    Czech Academy of Sciences Publication Activity Database

    Koloniuk, Igor; Hrabáková, Lenka; Petrzik, Karel

    2016-01-01

    Roč. 27, č. 6 (2016), s. 3979-3980 ISSN 1940 -1736 R&D Projects: GA MŠk LH13136; GA MŠk(CZ) EE2.3.30.0032 Institutional support: RVO:60077344 Keywords : Diaporthales * Mitogenome * Phomopsis Subject RIV: EB - Genetics ; Molecular Biology OBOR OECD: Genetics and heredity (medical genetics to be 3) Impact factor: 3.350, year: 2016

  15. A mitovirus isolated from the phytopathogenic fungus Alternaria brassicicola.

    Science.gov (United States)

    Chen, Yi; Shang, Hong Hong; Yang, Hong Qi; Da Gao, Bi; Zhong, Jie

    2017-09-01

    Alternaria brassicicola is one of the causal agents of alternaria blackspot in rapeseed. In this study, a dsRNA segment was isolated and sequenced from the fungus. The complete nucleotide sequence of the dsRNA was 2506 bp in length and, using the fungal mitochondrial genetic code, was predicted to contain a single large open reading frame (ORF) in the positive strand. This ORF was predicted to encode a protein with 719 amino acids that contains characteristic conserved motifs of the RNA-dependent RNA polymerase (RdRp). BLAST analysis revealed that this protein had significant sequence similarity to the RdRp from viruses of the genus Mitovirus. These results indicated that the dsRNA segment represents the replicative form of a mitovirus, which is tentatively designated "Alternaria brassicicola mitovirus 1" (AbMV1) and is a new member of the genus Mitovirus in the family Narnaviridae.

  16. Phytopathogenic bacteria in the system of modern agriculture.

    Science.gov (United States)

    Patyka, V P; Pasichnyk, L A

    2014-01-01

    The stages of studying bacterial diseases of crops and weeds at various farming systems have been characterized, biological properties have been investigated and pathogens identified using traditional and modern molecular genetic methods of research.

  17. In vitro screening of soil bacteria for inhibiting phytopathogenic fungi ...

    African Journals Online (AJOL)

    At present, the greatest interest resides with the development and application of specific biocontrol agent for the control of diseases on plant and this form the focus of this work. Several soil bacteria were evaluated in vitro for their effectiveness on the basis of their ability to suppress fungi in plate inhibition assays. 51 strains ...

  18. Antibacterial Activity of Phenolic Compounds Against the Phytopathogen Xylella fastidiosa

    OpenAIRE

    Maddox, Christina E.; Laur, Lisa M.; Tian, Li

    2010-01-01

    Xylella fastidiosa is a pathogenic bacterium that causes diseases in many crop species, which leads to considerable economic loss. Phenolic compounds (a group of secondary metabolites) are widely distributed in plants and have shown to possess antimicrobial properties. The anti-Xylella activity of 12 phenolic compounds, representing phenolic acid, coumarin, stilbene and flavonoid, was evaluated using an in vitro agar dilution assay. Overall, these phenolic compounds were effective in inhibiti...

  19. Alveolarides: Antifungal Peptides from Microascus alveolaris Active against Phytopathogenic Fungi.

    Science.gov (United States)

    Fotso, Serge; Graupner, Paul; Xiong, Quanbo; Gilbert, Jeffrey R; Hahn, Don; Avila-Adame, Cruz; Davis, George; Sumiyoshi, Kengo

    2018-01-26

    Three novel cyclodepsipeptides, alveolarides A (1), B (2), and C (3), each possessing the rare 2,3-dihydroxy-4-methyltetradecanoic acid unit and a β-phenylalanine amino acid residue, along with the known peptide scopularide were isolated and identified from the culture broth of Microascus alveolaris strain PF1466. The pure compounds were evaluated for biological activity, and alveolaride A (1) provided strong in vitro activity against the plant pathogens Pyricularia oryzae, Zymoseptoria tritici, and Ustilago maydis. Moderate activity of alveolaride A was observed under in planta conditions against Z. tritici, Puccinia triticina, and Phakopsora pachyrhizi. Structures of 1, 2, and 3 were determined by detailed analysis of NMR (1D and 2D) and mass spectrometry data. The partial absolute configuration of alveolaride A (1) was established.

  20. Enterobacter agglomerans: the clinically important plant pathogen.

    Science.gov (United States)

    Geere, I. W.

    1977-01-01

    During a 5-month period Enterobacter agglomerans, now described as a member of the phytopathogenic genus Erwinia, was isolated from 13 patients in a general hospital; in 1 patient it was isolated from two sites. In six instances the organism was the sole pathogen isolated, in two instances it may have contributed to infection and in the remaining instances it was probably a transient saprophyte. The strains showed some variation in biochemical reactions but were similar in colonial morphology and were consistently sensitive to several antibiotics. Although this organism is prevalent in the general environment and usually relatively benign, it does have a potential for nosocomial infection. PMID:837319

  1. Fitodefensivos em plantas medicinais: macromoléculas hidrofílicas de folhas de mil folhas (Achillea millefolium L. inibem o crescimento in vitro de bactérias fitopatogênicas Agrochemicals in medicinal plants: hydrophilic macromolecules from leaves of "mil folhas" (Achillea millefolium L. inhibit in vitro growth of phytopathogenic bacteria

    Directory of Open Access Journals (Sweden)

    N.G. Tessarollo

    2013-01-01

    Full Text Available Extratos aquosos da planta medicinal Achillea millefolium contêm macromoléculas de interesse para desenvolver fitodefensivos para a agricultura. Duas frações de mil folhas foram obtidas por ultrafiltração, E1 (contendo moléculas maiores que 30 kDa, e E3 (peptídeos entre 1 e 10 kDa que inibiram o crescimento das bactérias fitopatogênicas Ralstonia solanacearum, gram-negativa, e Clavibacter michiganensis subsp. michiganensis, gram-positiva, com dependência de concentração. Os valores de concentração inibitória mínima (CIM para ambos os extratos e bactérias foram baixos, entre 20 e 80µM. A CIM relativa à proteína total evidenciou a presença de macromoléculas muito ativas em E3, embora com baixa concentração proteica. E3 se aplica à prospecção de peptídeos antimicrobianos. Estimar a CIM relativa à quantidade de amostra vegetal valorizou o potencial antimicrobiano natural de E1, que contém alta concentração proteica. E1e E3 se aplicam ao desenvolvimento de fitodefensivos para uso biotecnológico. A ultrafiltração fracionou as amostras de forma nativa, rápida, e com baixo custo; além de dessalinizar, clarificar, purificar, e concentrar E1 e E3. Esse estudo inédito sobre a separômica e a ação antimicrobiana de extratos macromoleculares aquosos de mil folhas sugere que plantas cicatrizantes podem apresentar grande potencial para desenvolver fitodefensivos agrícolas naturais não danosos, à semelhança de medicamentos fitoterápicos.Aqueous extracts from the medicinal plant Achillea millefolium contain macromolecules of interest to develop agrochemicals for agriculture. Two fractions of "mil folhas" were obtained by ultrafiltration, E1 (containing molecules larger than 30 kDa and E3 (peptides between 1 and 10 kDa, which inhibited the growth of phytopathogenic bacteria Ralstonia solanacearum, gram-negative, and Clavibacter michiganensis subsp. michiganensis, gram-positive, concentration-dependent. The values of

  2. Antimicrobial activity of olive solutions from stored Alpeorujo against plant pathogenic microorganisms.

    Science.gov (United States)

    Medina, Eduardo; Romero, Concepcion; de Los Santos, Berta; de Castro, Antonio; Garcia, Aranzazu; Romero, Fernando; Brenes, Manuel

    2011-07-13

    The aim of this work was to assess the in vitro antimicrobial effects that wastewaters from alpeorujo oil extraction have against phytopathogenic bacteria and fungi. Alpeorujo was stored for 6 months and then processed to extract its oil, pomace, and a new liquid waste (OWSA), which was characterized by its content in phenolic compounds. OWSA at 20% decreased bu >4 log the population of Erwinia spp., Pseudomonas spp., and Clavibacter spp. viable cells in test tubes, whereas OWSA at 50% in agar medium was necessary to inhibit mycelial growth of most fungi. It was found that the bactericidal effect was due to the joint action of low molecular mass phenolic compounds, although neither hydroxytyrosol, its glucosides, hydroxytyrosol glycol, nor a glutaraldehyde-like compound individually explained this bioactivity. Hence, OWSA constitutes a promising natural solution to fight plant phytopathogenic bacteria and fungi.

  3. Seca de ponteiros de Eucalyptus spp. causada por Erwinia psidii no Brasil

    OpenAIRE

    Arriel, Daniele Aparecida Alvarenga

    2012-01-01

    A partir de 2009 vem sendo observada uma nova doença em eucalipto de etiologia bacteriana desconhecida, caracterizada por seca de ponteiros, murcha, lesões em ramos, pecíolo e nervura central, acompanhado de exsudação bacteriana macroscópica e microscópica. Até o presente a enfermidade já foi constatada em plantios clonais de híbridos de Eucalyptus urophylla x E. grandis, E. grandis e em plantios seminais de E. dunnii nos estados de São Paulo e Rio Grande do Sul. Devido à importância econômic...

  4. The Production, Purification and Properties of the Biopolymer Levan Produced by the Bacterium Erwinia Herbicola

    Science.gov (United States)

    1989-08-01

    after comparison with ATCC 11142 Acetobacter pasteurianus. ATCC 15953 Microbacterium 2 laevanlformans. and QMB 1624 Bacillus coagulans (B, gub^jlu...is produced by several bacteria, including Brwjnja herbicola. Streptococcus salivarius. Pseudomonas prunicola. Bacillus subtilis. and Actinomvcetes...J, i X I i rtfi ii i IT PTT-n T~^L^^^^ r1 •» ■ ■ 11 ■ ■ ■ i >■ 111 I’I I’.VIT fi-H-rtfi 1 -f ’ •■» i | i r , , , Levan from Bacillus

  5. Evaluation of three differential media for detection of Enterobacter agglomerans (Erwinia herbicola).

    Science.gov (United States)

    Bucher, C; von Graevenitz, A

    1982-01-01

    Dextrin-fuchsine-sulfite medium (DFS), Rimler-Shotts agar (RS), and a new lysine-ornithine-mannitol agar (LOM) were tested for detection of Enterobacter agglomerans. In human stools, LOM and DFS were most sensitive at coliform-to-E. agglomerans ratios of less than or equal to 10(2) and E. agglomerans inocula of greater than 10(2) per plate. Both LOM and DFS detected one strain in 254 stools, but RS proved to be inhibitory. PMID:6809787

  6. Evaluation of three differential media for detection of Enterobacter agglomerans (Erwinia herbicola).

    OpenAIRE

    Bucher, C; von Graevenitz, A

    1982-01-01

    Dextrin-fuchsine-sulfite medium (DFS), Rimler-Shotts agar (RS), and a new lysine-ornithine-mannitol agar (LOM) were tested for detection of Enterobacter agglomerans. In human stools, LOM and DFS were most sensitive at coliform-to-E. agglomerans ratios of less than or equal to 10(2) and E. agglomerans inocula of greater than 10(2) per plate. Both LOM and DFS detected one strain in 254 stools, but RS proved to be inhibitory.

  7. Epidemiology of Erwinia pyrifoliae, a new pathogen on strawberry in the Netherlands

    NARCIS (Netherlands)

    Wenneker, M.; Bergsma-Vlami, M.; Steen, van der J.J.M.

    2017-01-01

    During the late spring of 2013 strawberry plants (Fragaria × ananassa Duch. 'Elsanta') were showing an intense blackening of their immature fruits, their fruit calyxes and the peduncule were found at several locations (greenhouses) in The Netherlands. Symptoms include brown petals, green young

  8. Genetic modification of potato against microbial diseases: in vitro and in planta activity of a dermaseptin B1 derivative, MsrA2.

    Science.gov (United States)

    Osusky, Milan; Osuska, Lubica; Kay, William; Misra, Santosh

    2005-08-01

    Dermaseptin B1 is a potent cationic antimicrobial peptide found in skin secretions of the arboreal frog Phyllomedusa bicolor. A synthetic derivative of dermaseptin B1, MsrA2 (N-Met-dermaseptin B1), elicited strong antimicrobial activities against various phytopathogenic fungi and bacteria in vitro. To assess its potential for plant protection, MsrA2 was expressed at low levels (1-5 microg/g of fresh tissue) in the transgenic potato (Solanum tuberosum L.) cv. Desiree. Stringent challenges of these transgenic potato plants with a variety of highly virulent fungal phytopathogens--Alternaria, Cercospora, Fusarium, Phytophthora, Pythium, Rhizoctonia and Verticillium species--and with the bacterial pathogen Erwinia carotovora demonstrated that the plants had an unusually broad-spectrum and powerful resistance to infection. MsrA2 profoundly protected both plants and tubers from diseases such as late blight, dry rot and pink rot and markedly extended the storage life of tubers. Due to these properties in planta, MsrA2 is proposed as an ideal antimicrobial peptide candidate to significantly increase resistance to phytopathogens and improve quality in a variety of crops worldwide with the potential to obviate fungicides and facilitate storage under difficult conditions.

  9. Differences in the Distribution of Bi?fidobacterial and Enterobacterial Species in Human Faecal Microflora of Three Different (Children, Adults, Elderly) Age Groups

    OpenAIRE

    Gavini, Franҫoise; Cayuela, Chantal; Antoine, Jean-Michel; Lecoq, Catherine; Lefebvre, Bernard; Membré, Jeanne-Marie; Neut, Christel

    2011-01-01

    To determine if there is any change with age in the distribution of Bifidobacterium and Enterobacteriaceae species in human intestinal microflora, strains were isolated from a total of 54 samples of human faeces (15 children, 3–15 years old; 17 adults, 30–46 years old; 22 elderly, 69–89 years old). The bifidobacteria were identified at species level using a numerical phenotypic identification system developed on the Internet (http://kounou.lille.inra.fr) and DNA–DNA hy...

  10. Pesquisa de enterobacteriáceas e enterovírus em crianças normais e com quadros diarréicos agudos

    Directory of Open Access Journals (Sweden)

    José Alberto N. Candeias

    1968-12-01

    Full Text Available Num grupo de 263 crianças de 0 a 24 meses de idade, atendidas de outubro de 1963 a setembro de 1964 no Centro de Saúde da Lapa, na Capital de São Paulo, foram feitos exames de fezes para a pesquisa de Escherichia coli, do grupo da gastroenterite infantil, Shigella, Salmonella, poliovírus, vírus Coxsackie e vírus ECHO. O agente mais freqüentemente isolado foi o vírus da poliomielite (15,97% de positividade, seguindo-se-lhe a Escherichia coli G.E.I. (10,65%, Shigella (9,51%, vírus Coxsackie (7,22%, Salmonella em 3,04% dos casos e vírus ECHO no mesmo percentual. Em 16,22% das 111 que apresentaram resultado positivo, o exame revelou a presença de dois ou mais agentes. Os agentes acima referidos foram isolados de 42,21% da totalidade das crianças examinadas. Dentre as 167 que apresentavam diarréia, a porcentagem de positividade chegou a 47,90%, e nas 96 que nao a referiam, desceu a 32,29%. Sòmente em relação às shigelas foi possível evidenciar associação entre isolamento de microrganismos e quadro diarréico. Para a E. coli 0111, salmonelas e vírus Coxsackie A, os resultados foram apenas sugestivos desta associação, e para os demais agentes não houve evidência alguma favorável à mesma.A sample of 263 children between 0 and 24 months of age, among those attending the Lapa Health Center, São Paulo, Brazil, from October 1963 to September 1964 was studied. Cultures were performed for enteropathogenic Escherichia coli, Shigella, Salmonella, poliovirus, Coxsackie virus and ECHO virus in the feces. The commonest agent isolated was poliovirus (15.97% positive followed by enteropathogenic Escherichia coli (10.65%, Shigella (9.51%, Coxsackie (7.22% and finally Salmonella and ECHO (3.04% each. Of the 111 subjects that presented positive results, 16,22% were positive for two or more different agents. The above mentioned microorganisms were present in 42.21% of the children examined; of the 167 that presented diarrhea the percentage of positives was 47.90% and of the 96 that had no diarrhea, 32.29%. Association between diarrhea and the presence of an etiological agent was only demonstrated for the Shigella.Results were only suggestive for such association regarding E. coli 0111, Salmonella and Coxsackie A virus. Regarding the other agents there was no evidence of association.

  11. Isolation and 16s rdna sequence analysis of bacteria from dieback affected mango orchards in southern pakistan

    International Nuclear Information System (INIS)

    Khan, I.A.; Khan, A.; Asif, H.; Azim, M.K.; Muhlbach, H.P.

    2014-01-01

    A broad range of microorganisms are involved in various mango plant diseases such as fungi, algae and bacteria. In order to study the role of bacteria in mango dieback, a survey of infected mango plants in southern Pakistan was carried out. A number of bacterial isolates were obtained from healthy looking and infected mango trees, and their characterization was undertaken by colony PCR and subsequent sequence analysis of 16S rDNA. These analyses revealed the presence of various genera including Acinetobacter, Bacillus, Burkholderia, Cronobacter, Curtobacterium, Enterobacter, Erwinia, Exiguobacterium, Halotelea, Lysinibacillus, Micrococcus, Microbacterium, Pantoea, Pseudomonas, Salmonella and Staphylococcus. It is noteworthy that several members of these genera have been reported as plant pathogens. The present study provided baseline information regarding the phytopathogenic bacteria associated with mango trees in southern Pakistan. (author)

  12. Anthraquinones isolated from Cassia tora (Leguminosae) seed show an antifungal property against phytopathogenic fungi.

    Science.gov (United States)

    Kim, Young-Mi; Lee, Chi-Hoon; Kim, Hyo-Gyung; Lee, Hoi-Seon

    2004-10-06

    The fungicidal activities of Cassia tora extracts and their active principles were determined against Botrytis cineria, Erysiphe graminis, Phytophthora infestans, Puccinia recondita, Pyricularia grisea, and Rhizoctonia solani using a whole plant method in vivo and were compared with synthetic fungicides and three commercially available anthraquinones. The responses varied with the plant pathogen tested. At 1 g/L, the chloroform fraction of C. tora showed a strong fungicidal activity against B. cinerea, E. graminis, P. infestans, and R. solani. Emodin, physcion, and rhein were isolated from the chloroform fraction using chromatographic techniques and showed strong and moderate fungicidal activities against B. cinerea, E. graminis, P. infestans, and R. solani. Furthermore, aloe-emodin showed strong and moderate fungicidal activities against B. cinerea and R. solani, respectively, but did not inhibit the growth of E. graminis, P. infestans, P. recondita, and Py. grisea. Little or no activity was observed for anthraquinone and anthraquinone-2-carboxylic acid when tested at 1 g/L. Chlorothalonil and dichlofluanid as synthetic fungicides were active against P. infestans and B. cinerea at 0.05 g/L, respectively. Our results demonstrate the fungicidal actions of emodin, physcion, and rhein from C. tora.

  13. Suppression of phytopathogenic fungi by hexane extract of Nepenthes ventricosa x maxima leaf.

    Science.gov (United States)

    Shin, Kwang-Soo; Lee, Samkeun; Cha, Byeong Jin

    2007-12-01

    The hexane extract of Nepenthes ventricosa x maxima leaf exhibited antifungal activity against Alternaria alternata, Aspergillus niger, Bipolaris oryzae, Fusarium oxysporum, Phytophthora capsici, Rhizoctonia solani, Rhizopus stolonifer var. stolonifer and Sclerotinia sclerotiorum.

  14. Suppressive effects of metabolites from Photorhabdus spp. and Xenorhabdus spp. on phytopathogens of peach and pecan

    Science.gov (United States)

    Our objective was to determine the suppressive abilities of bacterial metabolites derived from Photorhabdus and Xenorhabdus spp. on Glomerella cingulata, Phomopsis sp., Phytophthora cactorum, and Fusicladosporium effusum, which are fungal or oomycete pathogens of pecan, and Monilinia fructicola, a f...

  15. Growth and enzymatic responses of phytopathogenic fungi to glucose in culture media and soil

    Directory of Open Access Journals (Sweden)

    Beatriz de Oliveira Costa

    2012-03-01

    Full Text Available The effect of inoculation of Aspergillus flavus, Fusarium verticillioides, and Penicillium sp. in Dystrophic Red Latosol (DRL and Eutroferric Red Latosol (ERL soils with or without glucose on the total carbohydrate content and the dehydrogenase and amylase activities was studied. The fungal growth and spore production in culture medium with and without glucose were also evaluated. A completely randomized design with factorial arrangement was used. The addition of glucose in the culture medium increased the growth rate of A. flavus and Penicillium sp. but not of F. verticillioides. The number of spores increased 1.2 for F. verticillioides and 8.2 times for A. flavus in the medium with glucose, but was reduced 3.5 times for Penicillium sp. The total carbohydrates contents reduced significantly according to first and second degree equations. The consumption of total carbohydrates by A. flavus and Penicillium sp. was higher than the control or soil inoculated with F. verticillioides. The addition of glucose to soils benefited the use of carbohydrates, probably due to the stimulation of fungal growth. Dehydrogenase activity increased between 1.5 to 1.8 times (p <0.05 in soils with glucose and inoculated with the fungi (except F. verticillioides, in relation to soil without glucose. Amylase activity increased 1.3 to 1.5 times due to the addition of glucose in the soil. Increased amylase activity was observed in the DRL soil with glucose and inoculated with A. flavus and Penicillium sp. when compared to control.

  16. Bacterial chitinase with phytopathogen control capacity from suppressive soil revealed by functional metagenomics.

    Science.gov (United States)

    Hjort, Karin; Presti, Ilaria; Elväng, Annelie; Marinelli, Flavia; Sjöling, Sara

    2014-03-01

    Plant disease caused by fungal pathogens results in vast crop damage globally. Microbial communities of soil that is suppressive to fungal crop disease provide a source for the identification of novel enzymes functioning as bioshields against plant pathogens. In this study, we targeted chitin-degrading enzymes of the uncultured bacterial community through a functional metagenomics approach, using a fosmid library of a suppressive soil metagenome. We identified a novel bacterial chitinase, Chi18H8, with antifungal activity against several important crop pathogens. Sequence analyses show that the chi18H8 gene encodes a 425-amino acid protein of 46 kDa with an N-terminal signal peptide, a catalytic domain with the conserved active site F175DGIDIDWE183, and a chitinase insertion domain. Chi18H8 was expressed (pGEX-6P-3 vector) in Escherichia coli and purified. Enzyme characterization shows that Chi18H8 has a prevalent chitobiosidase activity with a maximum activity at 35 °C at pH lower than 6, suggesting a role as exochitinase on native chitin. To our knowledge, Chi18H8 is the first chitinase isolated from a metagenome library obtained in pure form and which has the potential to be used as a candidate agent for controlling fungal crop diseases. Furthermore, Chi18H8 may also answer to the demand for novel chitin-degrading enzymes for a broad range of other industrial processes and medical purposes.

  17. Ancestral state reconstruction infers phytopathogenic origins of sooty blotch and flyspeck fungi on apple.

    Science.gov (United States)

    Ismail, Siti Izera; Batzer, Jean Carlson; Harrington, Thomas C; Crous, Pedro W; Lavrov, Dennis V; Li, Huanyu; Gleason, Mark L

    2016-01-01

    Members of the sooty blotch and flyspeck (SBFS) complex are epiphytic fungi in the Ascomycota that cause economically damaging blemishes of apples worldwide. SBFS fungi are polyphyletic, but approx. 96% of SBFS species are in the Capnodiales. Evolutionary origins of SBFS fungi remain unclear, so we attempted to infer their origins by means of ancestral state reconstruction on a phylogenetic tree built utilizing genes for the nuc 28S rDNA (approx. 830 bp from near the 59 end) and the second largest subunit of RNA polymerase II (RPB2). The analyzed taxa included the well-known genera of SBFS as well as non-SBFS fungi from seven families within the Capnodiales. The non-SBFS taxa were selected based on their distinct ecological niches, including plant-parasitic and saprophytic species. The phylogenetic analyses revealed that most SBFS species in the Capnodiales are closely related to plant-parasitic fungi. Ancestral state reconstruction provided strong evidence that plant-parasitic fungi were the ancestors of the major SBFS lineages. Knowledge gained from this study may help to better understand the ecology and evolution of epiphytic fungi. © 2016 by The Mycological Society of America.

  18. Dolabra nepheliae on rambutan and lychee represents a novel lineage of phytopathogenic Eurotiomycetes.

    Science.gov (United States)

    Rossman, Amy Y; Schoch, Conrad L; Farr, David F; Nishijima, Kate; Keith, Lisa; Goenaga, Ricardo

    2010-07-01

    Rambutan (Nephelium lappaceum) and lychee (Litchi chinensis) are tropical trees in the Sapindaceae that produce delicious edible fruits and are increasingly cultivated in tropical regions. These trees are afflicted with a stem canker disease associated with the ascomycete Dolabra nepheliae. Previously known from Asia and Australia, this fungus was recently reported from Hawaii and Puerto Rico. The sexual and asexual states of Dolabra nepheliae are redescribed and illustrated. In addition, the ITS and large subunit of the nuclear ribosomal DNA plus fragments from the genes RPB2, TEF1, and the mitochondrial small ribosomal subunit were sequenced for three isolates of D. nepheliae and compared with other sequences of ascomycetes. It was determined that D. nepheliae represents a new lineage within the Eurotiomycetes allied with Phaeomoniella chlamydospora, the causal agent of Petri grapevine decline.

  19. Rapid and efficient extraction of genomic DNA from different phytopathogenic fungi using DNAzol reagent.

    Science.gov (United States)

    Guo, Jian-Rong; Schnieder, F; Abd-Elsalam, K A; Verreet, J A

    2005-01-01

    A modified procedure using the commercial DNAzol reagent was successfully applied to extract genomic DNA from 25 fungal species. The DNA yield varied from 306 to 1,927 microg g(-1) dry mycelia and the A(260)/A(280) ratio from 1.59 to 1.93. Compared with the method of J.L. Cenis (Nucleic Acids Res. 1992, 20: 2380) this procedure generated a higher DNA yield from 17 species and a higher A(260)/A(280) ratio from 23 species. But for four species, Cenis (1992) method was more suitable. No inhibitor of polymerase chain reaction was evident for the DNA extracted by the modified procedure, whereas some inhibitors remained in DNA of eight species extracted by the previous method.

  20. Suppression of polygalacturonase gene expression in the phytopathogenic fungus Ophiostoma novo-ulmi by RNA interference.

    Science.gov (United States)

    Carneiro, Joyce S; de la Bastide, Paul Y; Chabot, Meghan; Lerch, Lindsey; Hintz, William E

    2010-05-01

    The fungal pathogen, Ophiostomo novo-ulmi, has been responsible for the rapid decline of American elm (Ulmus americana) across North America and remains a serious threat to surviving elm populations. The production of pectinolytic polygalacturonase enzymes has been implicated as a virulence factor for many fungal pathogens, including O. novo-ulmi. Previous work has shown that the targeted disruption of the endopolygalacturonase gene locus epg1 of O. novo-ulmi reduced, but did not eliminate pectinase activity. In the present study, we evaluated the use of RNA interference (RNAi) as a method of suppressing expression of the epg1 locus in O. novo-ulmi and compared its efficiency to the gene disruption method. While there was a reduction in epg1-specific mRNA transcripts and in the amount of polygalacturonase enzyme secreted for both methods of gene regulation, neither method completely suppressed the expression of pectinase activity. There was, however, a significantly greater reduction in both transcript levels and secreted enzyme observed for some of the RNAi transformants. As the first demonstration of RNAi in O. novo-ulmi, this method of gene regulation shows promise in future studies of gene expression and pathogenicity. Copyright 2010 Elsevier Inc. All rights reserved.

  1. The early response during the interaction of fungal phytopathogen and host plant.

    Science.gov (United States)

    Shen, Yilin; Liu, Na; Li, Chuang; Wang, Xin; Xu, Xiaomeng; Chen, Wan; Xing, Guozhen; Zheng, Wenming

    2017-05-01

    Plants can be infected by a variety of pathogens, most of which can cause severe economic losses. The plants resist the invasion of pathogens via the innate or acquired immune system for surviving biotic stress. The associations between plants and pathogens are sophisticated beyond imaging and the interactions between them can occur at a very early stage after their touching each other. A number of researchers in the past decade have shown that many biochemical events appeared even as early as 5 min after their touching for plant disease resistance response. The early molecular interactions of plants and pathogens are likely to involve protein phosphorylation, ion fluxes, reactive oxygen species (ROS) and other signalling transduction. Here, we reviewed the recent progress in the study for molecular interaction response of fungal pathogens and host plant at the early infection stage, which included many economically important crop fungal pathogens such as cereal rust fungi, tomato Cladosporium fulvum , rice blast and so on. By dissecting the earlier infection stage of the diseases, the avirulent/virulent genes of pathogen or resistance genes of plant could be defined more clearly and accurately, which would undoubtedly facilitate fungal pathogenesis study and resistant crop breeding. © 2017 The Authors.

  2. Phytochemistry, antioxidant potential and antifungal of Byrsonima crassifolia on soil phytopathogen control

    Directory of Open Access Journals (Sweden)

    B. S. Andrade

    2017-07-01

    Full Text Available Abstract The use of chemical defensives to control fungal diseases has by consequence to impact negatively over the environment and human health, this way, the use of plant extracts with antifungal properties along with proper cultural management makes viable an alternative plant production control, specially for familiar and organic cultures. The objective of this study was to perform phytochemical and antioxidant analysis of Byrsonima crassifolia (canjiqueira barks and evaluate its antifungal potential over Fusarium solani and Sclerotinia sclerotiorum mycelial growth. The ethanol extract from plants collected in Pantanal, Mato Grosso do Sul, Brazil was submitted to phytochemical prospection, total phenol and flavonoids quantification and antioxidant activiy determination (DPPH. To evaluate antifungal activity concentrations of 800, 1200, 1600, 2000 and 2400 µg 100 mL-1 of ethanol extract were used. Which concentration was separately incorporated in agar (PDA and shed in Petri dishes, followed by the fungi mycelial disc where the colonies diameter was measured daily. Negatives control with agar without extract and agar with an ethanol solution were used. The B. crassifolia ethanol extract presented inhibitory activity over the fungi studied where concentrations of 800 and 1600 µg 100 mL-1, inhibited 38% of the mycelial growth of F. solani; to S. sclerotiorum the best concentration was 2400 µg 100 mL1, reducing 37.5%. The antifungal bark extract potential of this specie is attributed to phenolic compounds and to triterpenes derivatives.

  3. Development of Conductive Polymer Analysis for the Rapid Detection and Identification of Phytopathogenic Microbes

    Science.gov (United States)

    A. Dan Wilson; D.G. Lester; C.S. Oberle

    2004-01-01

    Conductive polymer analysis, a type of electronic aroma detection technology, was evaluated for its efficacy in the detection, identification, and discrimination of plant-pathogenic microorganisms on standardized media and in diseased plant tissues. The method is based on the acquisition of a diagnostic electronic fingerprint derived from multisensor responses to...

  4. Volatile compounds emitted by diverse phytopathogenic microorganisms promote plant growth and flowering through cytokinin action

    Czech Academy of Sciences Publication Activity Database

    Sanchez-Lopez, A.; Baslam, M.; De Diego, N.; Jose Munoz, F.; Bahaji, A.; Almagro, G.; Ricarte-Bermejo, A.; Garcia-Gomez, P.; Li, J.; Humplík, J.F.; Novák, Ondřej; Spíchal, L.; Doležal, Karel; Baroja-Fernandez, E.; Pozueta-Romero, J.

    2016-01-01

    Roč. 39, č. 12 (2016), s. 2592-2608 ISSN 0140-7791 R&D Projects: GA MŠk(CZ) LO1204 Institutional support: RVO:61389030 Keywords : exceptionally high-levels * tandem mass-spectrometry * arabidopsis-thaliana * nitric-oxide * bacterial volatiles * floral transition * anthocyanin biosynthesis * transgenic arabidopsis * liquid-chromatography * organic-compounds * cytokinin * flowering * growth promotion * microbial volatile compounds * photoregulation * photosynthesis * plant-microbe interaction * starch Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 6.173, year: 2016

  5. Induction of infection in Teosinte (Zea diploperennis through the phytopathogen Ustilago maydis.

    Directory of Open Access Journals (Sweden)

    Martha Concepción Pérez Díaz

    2017-01-01

    Full Text Available Introduction: The corn and teosinte share morphological and molecular similarities latter being those that support the theory of teosinte (Zea parviglumis as its predecessor, both species are attacked by specific pathogens like Ustilago maydis. Objective: To analyze the infectious process that presents U. maydis on the variety of the teosinte Zea diploperennis. Materials and Methods: We used the strain of U. maydis FB-D12, which was kept a culture media rich in nutrients (CPES pH 7.0. Viable cells without morphological alterations to the inoculation method of puncture in teosinte seedlings were used. Monitoring of infection was carried out every 24 hours by measuring concentration of chlorophyll and plant tissue through microscopic observation Results: In the seedlings of Zea diploperennis inoculated with U. maydis the symptoms of the infection were presented, wilt and chlorosis in the leaves; The chlorosis was confirmed with the low concentration of chlorophyll 12 days later to the inoculation. In the microscopic observation of cuts of the tissue plant was found mycelium long and branched from the third day of the inoculation, until the appearance of tumors in seedlings of 45 days. Conclusions: The typical signs of infection with Ustilago maydis in the variety of teosinte Zea diploperennis do not differ from those reported for corn. Ustilago maydis presents its full life cycle within the plant, confirming that the diploperennis variety is susceptible.

  6. Trichoderma volatiles effecting Arabidopsis: from inhibition to protection against phytopathogenic fungi

    Directory of Open Access Journals (Sweden)

    Metwaly eRamadan

    2015-09-01

    Full Text Available Trichoderma species are present in many ecosystems and some strains have the ability to reduce the severity of plant diseases by activating various defence pathways via specific biologically active signaling molecules. Hence we investigated the effects of low molecular weight volatile compounds of Trichoderma asperellum IsmT5 on Arabidopsis thaliana. During co-cultivation of T. asperellum IsmT5 without physical contact to A. thaliana we observed smaller but vital and robust plants. The exposed plants exhibit increased trichome numbers, accumulation of defence-related compounds such as H2O2, anthocyanin, camalexin, and increased expression of defence related genes. We conclude that A. thaliana perceives the Trichoderma volatiles as stress compounds and subsequently initiates multilayered adaptations including activation of signaling cascades to withstand this environmental influence.The prominent headspace volatile of T. asperellum IsmT5 was identified to be 6-pentyl-α-pyrone, which was solely applied to A. thaliana to verify the growth and defence reactions. Most noticeable is that A. thaliana preexposed to 6PP showed significantly reduced symptoms when challenged with Botrytis cinerea and Alternaria brassicicola, indicating that defence-activated plants subsequently became more resistant to pathogen attack. Together, these results support that products that are based on Trichoderma volatiles have the potential being a useful biocontrol agent in agriculture.

  7. Designer phytoalexins: probing camalexin detoxification pathways in the phytopathogen Rhizoctonia solani.

    Science.gov (United States)

    Pedras, M Soledade C; Liu, Jun

    2004-04-07

    To probe the specificity of a camalexin detoxifying enzyme(s) produced by Rhizoctonia solani, the putative 5-camalexin hydroxylase (5-CAHY), the naturally occurring phytoalexin 1-methylcamalexin and designer phytoalexins in which the H-5 of camalexin was replaced with either a methyl group or a fluorine atom were synthesised. This investigation showed that biotransformation of 5-fluorocamalexin by R. solani was substantially slower than that of camalexin (12 days vs. six to eight hours), 5-methylcamalexin (5-6 days) or 1-methylcamalexin (5-6 days). Antifungal bioassays showed that 5-fluorocamalexin, 5-methylcamalexin and 1-methylcamalexin were more inhibitory to R. solani than camalexin, whereas their metabolic products displayed substantially lower inhibitory activity. It was concluded that detoxification via oxidation of the indole moiety of camalexins is predominant in the biotransformation of both camalexin and 5-methylcamalexin and likely catalysed by a specific 5-CAHY. By contrast, the pathways for detoxification of 1-methylcamalexin and 5-fluorocamalexin are likely catalysed by non-specific "house-keeping" enzymes. Most importantly, because 1- methylcamalexin showed stronger antifungal activity and was metabolised at substantially slower rate than camalexin this work suggested that, from a plant's perspective 1-methylcamalexin could be a more effective antifungal defence than camalexin.

  8. Peltaster fructicola genome reveals evolution from an invasive phytopathogen to an ectophytic parasite

    OpenAIRE

    Chao Xu; Huan Chen; Mark L. Gleason; Jin-Rong Xu; Huiquan Liu; Rong Zhang; Guangyu Sun

    2016-01-01

    Sooty blotch and flyspeck (SBFS) fungi are unconventional plant pathogens that cause economic losses by blemishing the surface appearance of infected fruit. Here, we introduce the 18.14-Mb genome of Peltaster fructicola, one of the most prevalent SBFS species on apple. This undersized assembly contains only 8,334 predicted protein-coding genes and a very small repertoire of repetitive elements. Phylogenomics and comparative genomics revealed that P. fructicola had undergone a reductive evolut...

  9. Essential oils from Algerian species of Mentha as new bio-control agents against phytopathogen strains.

    Science.gov (United States)

    Benomari, Fatima Zahra; Andreu, Vanessa; Kotarba, Jules; Dib, Mohammed El Amine; Bertrand, Cédric; Muselli, Alain; Costa, Jean; Djabou, Nassim

    2017-09-02

    Chemical composition and antifungal activity of essential oils of Algerian Mentha species were studied. Chemical compositions of different Mentha species oils (Mentha rotundifolia, M. spicata, M. pulegium, and M. piperita) were investigated by capillary GC and GC/MS, and their antifungal activities were evaluated by means of paper disc diffusion method and minimum inhibitory concentration (MIC) assays. In total, 98 components from all Mentha species were identified. All oils were rich in monoterpene-oxygenated components. In addition, we reported fumigant antifungal activity of Algerian Mentha essential oils against four fungi: Botrytis cinerea, Penicillium expansum, Monilinia laxa, and M. fructigena. All oils demonstrated very good inhibition especially against B. cinerea, M. laxa, and M. fructigena. Both Monilinia fungi were extremely sensitive to all Algerian Mentha oils, which suggests that Mentha essential oils have the potential to be used as bio-pesticides to protect fruit trees, such as apple and pear trees, and provides an alternative to chemical pesticides.

  10. Fumigant antifungal activity of essential oil components from Acorus gramineus against three phytopathogenic fungi.

    Science.gov (United States)

    Lee, Yeon-Suk; Kim, Junheon; Lee, Sang-Gil; Shin, Sang-Chul; Park, Il-Kwon

    2008-01-01

    Analysis by gas chromatography-mass spectrometry led to the identification of 26 compounds in Acorus gramineus essential oil. The antifungal activity of the identified compounds was tested singularly by using standard compounds. Allyl isothiocyanate and cis-asarone showed inhibition rates of 100% against P. cactorum at 28 mg/l air. In a test with C. parasitica and E. circinatum, allyl isothiocyante and cis-asarone showed moderate activity at 28 mg/l air.

  11. Ancestral state reconstruction infers phytopathogenic origins of sooty blotch and flyspeck fungi on apple

    NARCIS (Netherlands)

    Ismail, Siti Izera; Batzer, Jean Carlson; Harrington, Thomas C.; Crous, Pedro W.; Lavrov, Dennis V.; Li, Huanyu; Gleason, Mark L.

    2016-01-01

    Members of the sooty blotch and flyspeck (SBFS) complex are epiphytic fungi in the Ascomycota that cause economically damaging blemishes of apples worldwide. SBFS fungi are polyphyletic, but approx. 96% of SBFS species are in the Capnodiales. Evolutionary origins of SBFS fungi remain unclear, so we

  12. Ancestral state reconstruction infers phytopathogenic origins of sooty blotch and flyspeck fungi on apple

    NARCIS (Netherlands)

    Ismail, Siti Izera; Batzer, Jean Carlson; Harrington, Thomas C.; Crous, Pedro W.; Lavrov, Dennis V.; Li, Huanyu; Gleason, Mark L.

    2016-01-01

    Members of the sooty blotch and flyspeck (SBFS) complex are epiphytic fungi in the Ascomycota that cause economically damaging blemishes of apples worldwide. SBFS fungi are polyphyletic, but approx. 96% of SBFS species are in the Capnodiales. Evolutionary origins of SBFS fungi remain unclear, so

  13. Peltaster fructicola genome reveals evolution from an invasive phytopathogen to an ectophytic parasite.

    Science.gov (United States)

    Xu, Chao; Chen, Huan; Gleason, Mark L; Xu, Jin-Rong; Liu, Huiquan; Zhang, Rong; Sun, Guangyu

    2016-03-11

    Sooty blotch and flyspeck (SBFS) fungi are unconventional plant pathogens that cause economic losses by blemishing the surface appearance of infected fruit. Here, we introduce the 18.14-Mb genome of Peltaster fructicola, one of the most prevalent SBFS species on apple. This undersized assembly contains only 8,334 predicted protein-coding genes and a very small repertoire of repetitive elements. Phylogenomics and comparative genomics revealed that P. fructicola had undergone a reductive evolution, during which the numbers of orphan genes and genes involved in plant cell wall degradation, secondary metabolism, and secreted peptidases and effectors were drastically reduced. In contrast, the genes controlling 1,8-dihydroxynaphthalene (DHN)-melanin biosynthesis and appressorium-mediated penetration were retained substantially. Additionally, microscopic examination of the surfaces of infected apple indicated for the first time that P. fructicola can not only dissolve epicuticular waxes but also partially penetrate the cuticle proper. Our findings indicate that genome contraction, characterized mainly by the massive loss of pathogenicity-related genes, has played an important role in the evolution of P. fructicola (and by implication other SBFS species) from a plant-penetrating ancestor to a non-invasive ectophyte, displaying a novel form of trophic interaction between plants and fungi.

  14. Production of extracellular lipase by the phytopathogenic fungus Fusarium solani FS1

    OpenAIRE

    Maia, Maria de Mascena Diniz; Morais, Marcia Maria Camargo de; Morais Jr., Marcos Antonio de; Melo, Eduardo Henrique Magalhães; Lima Filho, José Luiz de

    1999-01-01

    A Brazilian strain of Fusarium solani was tested for extracellular lipase production in peptone-olive oil medium. The fungus produced 10,500 U.l-1 of lipase after 72 hours of cultivation at 25oC in shake-flask at 120 rpm in a medium containing 3% (w/v) peptone plus 0.5% (v/v) olive oil. Glucose (1% w/v) was found to inhibit the inductive effect of olive oil. Peptone concentrations below 3% (w/v) resulted in a reduced lipase production while increased olive oil concentration (above 0.5%) did n...

  15. Fungal secondary metabolites as inhibitors of infection-related morphogenesis in phytopathogenic fungi.

    Science.gov (United States)

    Thines, Eckhard; Anke, Heidrun; Weber, Roland W S

    2004-01-01

    The life-cycle of many plant-pathogenic fungi, especially those infecting aerial plant organs, contains several specific developmental stages. If these are sufficiently distinct in their physiology from vegetative hyphal growth, they present potential targets for non-fungitoxic plant protectants. The present review identifies such targets especially in the pre-penetration stages of the infection cycle of Magnaporthe grisea and other fungi infecting from air-borne spores. Examples of non-toxic natural products with activity against spore germination, attachment, appressorium formation, appressorium maturation and penetration of the host surface are given. In contrast, no substances selectively active against in planta growth or sporulation appear to be known. The selective activity of numerous secondary metabolites against specific infection stages without accompanying toxicity against vegetatively growing hyphae indicates a direction for the development of future natural product-derived fungicides which are more easily degraded in the environment and possess fewer non-target effects. Such substances are produced by many saprotrophic and endophytic fungi in pure culture. The paucity of data on the production of biologically active substances in natural situations limits the interpretation of their ecophysiological significance for the producer.

  16. Optimization for rapid synthesis of silver nanoparticles and its effect on phytopathogenic fungi

    Science.gov (United States)

    Krishnaraj, C.; Ramachandran, R.; Mohan, K.; Kalaichelvan, P. T.

    In this present study, silver nanoparticles were synthesized by green chemistry approach using Acalypha indica leaf extract as reducing agents. The reaction medium employed in the synthesis process was optimized to attain better yield, controlled size and stability. Further, the biosynthesized silver nanoparticles were conformed through UV-vis spectrum, XRD and HR-TEM analyses. Different concentration of silver nanoparticles were tested to know the inhibitory effect of fungal plant pathogens namely Alternaria alternata, Sclerotinia sclerotiorum, Macrophomina phaseolina, Rhizoctonia solani, Botrytis cinerea and Curvularia lunata. Interestingly, 15 mg concentration of silver nanoparticles showed excellent inhibitory activity against all the tested pathogens. Thus, the obtained results clearly suggest that silver nanoparticles may have important applications in controlling various plant diseases caused by fungi.

  17. Synthesis, Biological Evaluation, and Molecular Modeling Studies of New Oxadiazole-Stilbene Hybrids against Phytopathogenic Fungi

    Science.gov (United States)

    Jian, Weilin; He, Daohang; Song, Shaoyun

    2016-08-01

    Natural stilbenes (especially resveratrol) play important roles in plant protection by acting as both constitutive and inducible defenses. However, their exogenous applications on crops as fungicidal agents are challenged by their oxidative degradation and limited availability. In this study, a new class of resveratrol-inspired oxadiazole-stilbene hybrids was synthesized via Wittig-Horner reaction. Bioassay results indicated that some of the compounds exhibited potent fungicidal activity against Botrytis cinerea in vitro. Among these stilbene hybrids, compounds 11 showed promising inhibitory activity with the EC50 value of 144.6 μg/mL, which was superior to that of resveratrol (315.6 μg/mL). Remarkably, the considerably abnormal mycelial morphology was observed in the presence of compound 11. The inhibitory profile was further proposed by homology modeling and molecular docking studies, which showed the possible interaction of resveratrol and oxadiazole-stilbene hybrids with the cytochrome P450-dependent sterol 14α-demethylase from B. cinerea (BcCYP51) for the first time. Taken together, these results would provide new insights into the fungicidal mechanism of stilbenes, as well as an important clue for biology-oriented synthesis of stilbene hybrids with improved bioactivity against plant pathogenic fungi in crop protection.

  18. Plant-phytopathogen interactions: bacterial responses to environmental and plant stimuli.

    Science.gov (United States)

    Leonard, Simon; Hommais, Florence; Nasser, William; Reverchon, Sylvie

    2017-05-01

    Plant pathogenic bacteria attack numerous agricultural crops, causing devastating effects on plant productivity and yield. They survive in diverse environments, both in plants, as pathogens, and also outside their hosts as saprophytes. Hence, they are confronted with numerous changing environmental parameters. During infection, plant pathogens have to deal with stressful conditions, such as acidic, oxidative and osmotic stresses; anaerobiosis; plant defenses; and contact with antimicrobial compounds. These adverse conditions can reduce bacterial survival and compromise disease initiation and propagation. Successful bacterial plant pathogens must detect potential hosts and also coordinate their possibly conflicting programs for survival and virulence. Consequently, these bacteria have a strong and finely tuned capacity for sensing and responding to environmental and plant stimuli. This review summarizes our current knowledge of the signals and genetic circuits that affect survival and virulence factor expression in three important and well-studied plant pathogenic bacteria with wide host ranges and the capacity for long-term environmental survival. These are: Ralstonia solanacerarum, a vascular pathogen that causes wilt disease; Agrobacterium tumefaciens, a biotrophic tumorigenic pathogen responsible for crown gall disease and Dickeya, a brute force apoplastic pathogen responsible for soft-rot disease. © 2016 Society for Applied Microbiology and John Wiley & Sons Ltd.

  19. Maize ZmRACK1 Is Involved in the Plant Response to Fungal Phytopathogens

    Directory of Open Access Journals (Sweden)

    Baosheng Wang

    2014-05-01

    Full Text Available The receptor for activated C kinase 1 (RACK1 belongs to a protein subfamily containing a tryptophan-aspartic acid-domain (WD repeat structure. Compelling evidence indicates that RACK1 can interact with many signal molecules and affect different signal transduction pathways. In this study, we cloned a maize RACK1 gene (ZmRACK1 by RT-PCR. The amino acid sequence of ZmRACK1 had seven WD repeats in which there were typical GH (glycine-histidine and WD dipeptides. Comparison with OsRACK1 from rice revealed 89% identity at the amino acid level. Expression pattern analysis by RT-PCR showed that ZmRACK1 was expressed in all analyzed tissues of maize and that its transcription in leaves was induced by abscisic acid and jasmonate at a high concentration. Overexpression of ZmRACK1 in maize led to a reduction in symptoms caused by Exserohilum turcicum (Pass. on maize leaves. The expression levels of the pathogenesis-related protein genes, PR-1 and PR-5, increased 2.5–3 times in transgenic maize, and reactive oxygen species production was more active than in the wild-type. Yeast two-hybrid assays showed that ZmRACK1 could interact with RAC1, RAR1 and SGT1. This study and previous work leads us to believe that ZmRACK1 may form a complex with regulators of plant disease resistance to coordinate maize reactions to pathogens.

  20. Potential Pseudomonas Isolated from Soybean Rhizosphere as Biocontrol against Soilborne Phytopathogenic Fungi

    Directory of Open Access Journals (Sweden)

    ARI SUSILOWATI

    2011-06-01

    Full Text Available Plants are liable to be attacked by soilborne fungal pathogens which are responsible to reduce plant growth and losses in yield. In Indonesia, indigenous soybeans’ rhizobacteria such as antifungal producing Pseudomonas sp. have not many been reported yet. Therefore, the potential of the Pseudomonas sp. as biocontrol agent should be deeply explored. The aim of this study was to screen the indigenous soybeans’ rhizobacteria Pseudomonas sp. that possessing biocontrol characters against soilborne mainly i.e. Sclerotium rolfsii, Fusarium oxysporum, and Rhizoctonia solani, in vitro and in planta. Eleven isolates identified Pseudomonas sp. CRB numbered by CRB-3, CRB-16, CRB-17, CRB-31, CRB-44, CRB-75, CRB-80, CRB-86, CRB-102, CRB-109, and CRB-112 were affirmed to be candidates of biocontrol agents toward the soilborne fungal pathogens. Pseudomonas sp. CRB inhibited growth of the pathogenic fungi approximately 11.1-60.0% in vitro. Among of them, 7 isolates were also produced siderophore, 2 isolates produced chitinase, and 4 isolates produced hydrogen cyanide. Seed coating with the Pseudomonas sp. CRB accomplished disease suppression in planta about 14.3-100% in sterile soil condition and 5.2-52.6% in non sterile soil condition. Consistency in high performance more than 30% of disease suppression in non sterile soil condition suggested that 5 isolates i.e. CRB-16, CRB-44, CRB-86, CRB-102, and CRB-109 isolates have great promising to be developed as biocontrol agents of soilborne pathogenic fungi.

  1. Molecular Genetic Methods Implementation for Phytopathogen Identification in Forest Stands and Nurseries of the Russian Federation

    Directory of Open Access Journals (Sweden)

    T. S. Alimova

    2014-08-01

    Full Text Available The results of the application of molecular genetics methods for the analysis of the plant pathogens present in forest plantations and nurseries of the Russian Federation, including doughnut fungus and annosum root rot are presented. The prospects and benefits of using DNA analysis for early diagnosis of plant diseases without isolation of the pathogen in pure culture, shortening time of analysis, and the possibility of mass screening are discussed.

  2. Gall-ID: tools for genotyping gall-causing phytopathogenic bacteria

    Directory of Open Access Journals (Sweden)

    Edward W. Davis II

    2016-07-01

    Full Text Available Understanding the population structure and genetic diversity of plant pathogens, as well as the effect of agricultural practices on pathogen evolution, is important for disease management. Developments in molecular methods have contributed to increase the resolution for accurate pathogen identification, but those based on analysis of DNA sequences can be less straightforward to use. To address this, we developed Gall-ID, a web-based platform that uses DNA sequence information from 16S rDNA, multilocus sequence analysis and whole genome sequences to group disease-associated bacteria to their taxonomic units. Gall-ID was developed with a particular focus on gall-forming bacteria belonging to Agrobacterium, Pseudomonas savastanoi, Pantoea agglomerans, and Rhodococcus. Members of these groups of bacteria cause growth deformation of plants, and some are capable of infecting many species of field, orchard, and nursery crops. Gall-ID also enables the use of high-throughput sequencing reads to search for evidence for homologs of characterized virulence genes, and provides downloadable software pipelines for automating multilocus sequence analysis, analyzing genome sequences for average nucleotide identity, and constructing core genome phylogenies. Lastly, additional databases were included in Gall-ID to help determine the identity of other plant pathogenic bacteria that may be in microbial communities associated with galls or causative agents in other diseased tissues of plants. The URL for Gall-ID is http://gall-id.cgrb.oregonstate.edu/.

  3. The Influence of Learning on Host Plant Preference in a Significant Phytopathogen Vector, Diaphorina citri.

    Directory of Open Access Journals (Sweden)

    Dara G Stockton

    Full Text Available Although specialist herbivorous insects are guided by innate responses to host plant cues, host plant preference may be influenced by experience and is not dictated by instinct alone. The effect of learning on host plant preference was examined in the Asian citrus psyllid, Diaphorina citri; vector of the causal agent of citrus greening disease or huanglongbing. We investigated: a whether development on specific host plant species influenced host plant preference in mature D. citri; and b the extent of associative learning in D. citri in the form of simple and compound conditioning. Learning was measured by cue selection in a 2-choice behavioral assay and compared to naïve controls. Our results showed that learned responses in D. citri are complex and diverse. The developmental host plant species influenced adult host plant preference, with female psyllids preferring the species on which they were reared. However, such preferences were subject to change with the introduction of an alternative host plant within 24-48 hrs, indicating a large degree of experience-dependent response plasticity. Additionally, learning occurred for multiple sensory modalities where novel olfactory and visual environmental cues were associated with the host plant. However, males and females displayed differing discriminatory abilities. In compound conditioning tasks, males exhibited recognition of a compound stimulus alone while females were capable of learning the individual components. These findings suggest D. citri are dynamic animals that demonstrate host plant preference based on developmental and adult experience and can learn to recognize olfactory and visual host plant stimuli in ways that may be sex specific. These experience-based associations are likely used by adults to locate and select suitable host plants for feeding and reproduction and may suggest the need for more tailored lures and traps, which reflect region-specific cultivars or predominate Rutaceae in the area being monitored.

  4. In vitro antifungal activity of Ocimum selloi essential oil and methylchavicol against phytopathogenic fungi1

    Directory of Open Access Journals (Sweden)

    Larissa Corrêa Bomfim Costa

    Full Text Available The efficacy of Ocimum selloi essential oil was evaluated for controlling the growth of mycelia and spores germination Moniliophthora perniciosa. Six compounds (99.89% of the total oil were identified by GC-MS, of which methyl chavicol, methyl eugenol, β-caryophyllene, germacrene-D, bicyclogermacrene and spathulenol. Essential oil was tested for antifungal activity, which was determined by disc diffusion and minimum inhibitory concentration (MIC determination methods. Application of the oil reduced mycelial growth in a dose dependent manner, with maximum inhibition being observed at concentration of 1,000 ppm. Such antifungal activity could be attributed to methyl chavicol since the pure compound was shown to be similarly effective against Moniliophthora perniciosa at 1,000 ppm. The oil when applied at a concentration of 1,000 ppm, reduced the spore germination of Colletotrichum gloeosporioides and M. perniciosa by 93 and 87%, respectively, but had no effect on the Alternaria alternata. It is concluded that the oil from O. selloi and its major constituent, methyl chavicol, are efficient in inhibiting M. perniciosa, but less effective against C. gloeosporioides and A. alternata. The results obtained from this work may contribute to the development of alternative anti-fungal agents to protect the cacao crop from fungal disease.

  5. Quantitative assessment of phytopathogenic fungi in various substrates using a DNA macroarray

    NARCIS (Netherlands)

    Lievens, B.; Brouwer, M.; Vanachter, A.C.R.C.; Lévesque, C.A.; Cammue, B.P.A.; Thomma, B.P.H.J.

    2005-01-01

    Detection, identification and quantification of plant pathogens are the cornerstones of preventive plant disease management. To detect multiple pathogens in a single assay, DNA array technology currently is the most suitable technique. However, for sensitive detection, polymerase chain reaction

  6. Unraveling the Protein Network of Tomato Fruit in Response to Necrotrophic Phytopathogenic Rhizopus nigricans

    Science.gov (United States)

    Pan, Xiaoqi; Zhu, Benzhong; Luo, Yunbo; Fu, Daqi

    2013-01-01

    Plants are endowed with a sophisticated defense mechanism that gives signals to plant cells about the immediate danger from surroundings and protects them from pathogen invasion. In the search for the particular proteins involved in fruit defense responses, we report here a comparative analysis of tomato fruit (Solanum lycopersicum cv. Ailsa Craig) infected by Rhizopus nigricans Ehrenb, which is a significant contributor to postharvest rot disease in fresh tomato fruits. In total, four hundred forty-five tomato proteins were detected in common between the non-infected group and infected tomato fruit of mature green. Forty-nine differentially expressed spots in 2-D gels were identified, and were sorted into fifteen functional groups. Most of these proteins participate directly in the stress response process, while others were found to be involved in several equally important biological processes: protein metabolic process, carbohydrate metabolic process, ethylene biosynthesis, and cell death and so on. These responses occur in different cellular components, both intra- and extracellular spaces. The differentially expressed proteins were integrated into several pathways to show the regulation style existing in tomato fruit host. The composition of the collected proteins populations and the putative functions of the identified proteins argue for their roles in pathogen-plant interactions. Collectively results provide evidence that several regulatory pathways contribute to the resistance of tomato fruit to pathogen. PMID:24023804

  7. Leaf-cutting ant fungi produce cell wall degrading pectinase complexes reminiscent of phytopathogenic fungi

    DEFF Research Database (Denmark)

    Schiøtt, Morten; Rogowska-Wrzesinska, Adelina; Roepstorff, Peter

    2010-01-01

    Leaf-cutting (attine) ants use their own fecal material to manure fungus gardens, which consist of leaf material overgrown by hyphal threads of the basidiomycete fungus Leucocoprinus gongylophorus that lives in symbiosis with the ants. Previous studies have suggested that the fecal droplets conta...

  8. Chitosan effects on phytopathogenic fungi and seed germination of Jatropha curcas L.

    Directory of Open Access Journals (Sweden)

    Diana Pabón-Baquero

    2015-01-01

    Full Text Available Jatropha curcas es una planta con gran potencial agrícola e industrial. En este estudio se aislaron dos hongos de semillas no germinadas. Los aislamientos fúngicos se identificaron morfológica y molecularmente como Fusarium equiseti y Curvularia lunata. Los efectos del quitosano se evaluaron sobre el crecimiento micelial, esporulación y germinación de esporas de F. equiseti y C. lunata. Además, se estudió el efecto sobre la germinación de las semillas de J. curcas. Los resultados demostraron que todas las concentraciones probadas de quitosano (0.5, 1.0, 2.0 y 4.0 mg·mL-1 inhibieron el crecimiento micelial de los hongos. Las respuestas de esporulación y germinación de esporas fueron diferentes dependiendo de la especie fúngica; el quitosano inhibió completamente la esporulación C. lunata y la germinación de esporas de F. equiseti. La inoculación con F. equiseti y C. lunata redujo la germinación de semillas de J. curcas 20 y 26.6 %, respectivamente; sin embargo, la aplicación de quitosano antes de la inoculación inhibió la actividad patogénica. En conclusión, el quitosano no afectó la germinación de las semillas y causó efectos inhibitorios en F. equiseti y C. lunata. Este es el primer reporte del efecto del quitosano en J. curcas.

  9. Exploring Western Ghats microbial diversity for antagonistic microorganisms against fungal phytopathogens of pepper and chickpea

    Directory of Open Access Journals (Sweden)

    B.N. RAMKUMAR

    2015-08-01

    Full Text Available Newly isolated microbial cultures from Western Ghat soil samples of Kerala region in India were screened for antagonistic activity by well diffusion and dual culture plating against Phytophthora capsici and Rhizoctonia solani, infecting pepper and chickpea, respectively. Bioactive samples were made by varying solvent extraction of the culture broths of the potent isolates belongs to Actinomycetes, Pseudomonas, Bacillus and Trichoderma. The efficacy of the isolates to produce other potent antifungal metabolites such as cell wall degrading enzymes, HCN and volatile compounds were also checked. Treatment with antagonistic isolates in vivo under greenhouse conditions revealed significant reduction of the disease intensity of foot rot disease of black pepper and collar rot of chick pea.

  10. Synthetic blends of volatile, phytopathogen-induced odorants can be used to manipulate vector behavior.

    Directory of Open Access Journals (Sweden)

    Alexander eAksenov

    2014-12-01

    Full Text Available Volatile organic compounds (VOCs are emitted from all plants and these VOCs are important means of communication between plants and insects. It has been documented that pathogen infections alter VOC profiles rendering infected plants more attractive to specific vectors transmitting these pathogens than uninfected plants, thus potentially aiding in pathogen propagation. Mimicking these chemical cues might enable insect attraction away from the plant or disruption of host finding behavior of the vector. However, the practical implications have not been fully explored. We used citrus, Diaphorina citri and huanglongbing (HLB as a model host-vector-disease system because HLB threatens citrus production worldwide and is similar to other critical diseases of food crops, such as Zebra Chip affecting potato. We formulated a synthetic chemical blend using selected HLB-specific biomarker compounds, and tested the blend with the Attenu assay system for chemosensory proteins. The Attenu assay system is a procedure that identifies interactions between insect chemosensory proteins and their ligands. We found that an equimolar mixture of compounds mimicking the volatile profile of HLB-infected citrus bound chemosensory proteins. Further investigation of this blend in laboratory behavioral assays resulted in development of a synthetic lure that was more attractive to D. citri than natural citrus tree volatiles. This strategy could provide a new route to produce chemical lures for vector population control for a variety of plant and/or animal systems and it may result in the development of a practical lure for monitoring vectors of disease, such as D. citri.

  11. Fungicidal property of Curcuma longa L. rhizome-derived curcumin against phytopathogenic fungi in a greenhouse.

    Science.gov (United States)

    Kim, Moo-Key; Choi, Gyung-Ja; Lee, Hoi-Seon

    2003-03-12

    Fungicidal activity of Curcuma longa rhizome-derived materials against Botrytis cineria, Erysiphe graminis, Phytophthora infestans, Puccinia recondita, Pyricularia oryzae, and Rhizoctonia solani was tested using a whole plant method in vivo. It was compared with synthetic fungicides and four commercially available compounds derived from C. longa. The response varied with the tested plant pathogen. At 1000 mg/L, the hexane extract of C. longa showed fungicidal activities against E.graminis, P. infestans, and R. solani, and the ethyl acetate extract of C. longa showed fungicidal activities against B. cineria, P. infestans, Pu. recondita, and R. solani. Curcumin was isolated from the ethyl acetate fraction using chromatographic techniques and showed fungicidal activities against P. infestans, Pu. recondita, and R. solani with 100, 100, and 63% control values at 500 mg/L and 85, 76, and 45% control values at 250 mg/L, respectively. In the test with components derived from C. longa, turmerone exhibited weak activity against E. graminis, but no activity was observed from treatments with borneol, 1,8-cineole, sabinene, and turmerone. In comparison, potent fungicidal activity with chlorothalonil against P. infestans at 50 mg/L and dichlofluanid against B. cinerea at 50 mg/L was exhibited. These results may be an indication of at least one of the fungicidal actions of curcumin derived from C. longa.

  12. Genome structure and reproductive behaviour influence the evolutionary potential of a fungal phytopathogen.

    Directory of Open Access Journals (Sweden)

    Guillaume Daverdin

    Full Text Available Modern agriculture favours the selection and spread of novel plant diseases. Furthermore, crop genetic resistance against pathogens is often rendered ineffective within a few years of its commercial deployment. Leptosphaeria maculans, the cause of phoma stem canker of oilseed rape, develops gene-for-gene interactions with its host plant, and has a high evolutionary potential to render ineffective novel sources of resistance in crops. Here, we established a four-year field experiment to monitor the evolution of populations confronted with the newly released Rlm7 resistance and to investigate the nature of the mutations responsible for virulence against Rlm7. A total of 2551 fungal isolates were collected from experimental crops of a Rlm7 cultivar or a cultivar without Rlm7. All isolates were phenotyped for virulence and a subset was genotyped with neutral genetic markers. Virulent isolates were investigated for molecular events at the AvrLm4-7 locus. Whilst virulent isolates were not found in neighbouring crops, their frequency had reached 36% in the experimental field after four years. An extreme diversity of independent molecular events leading to virulence was identified in populations, with large-scale Repeat Induced Point mutations or complete deletion of AvrLm4-7 being the most frequent. Our data suggest that increased mutability of fungal genes involved in the interactions with plants is directly related to their genomic environment and reproductive system. Thus, rapid allelic diversification of avirulence genes can be generated in L. maculans populations in a single field provided that large population sizes and sexual reproduction are favoured by agricultural practices.

  13. Rhizospheric fungi of Panax notoginseng: diversity and antagonism to host phytopathogens

    Directory of Open Access Journals (Sweden)

    Cui-Ping Miao

    2016-04-01

    Conclusion: Our results suggest that diverse fungi including potential pathogenic ones exist in the rhizosphere soil of 2-yr-old P. notoginseng and that antagonistic isolates may be useful for biological control of pathogens.

  14. Phytochemistry, antioxidant potential and antifungal of Byrsonima crassifolia on soil phytopathogen control.

    Science.gov (United States)

    Andrade, B S; Matias, R; Corrêa, B O; Oliveira, A K M; Guidolin, D G F; Roel, A R

    2018-02-01

    The use of chemical defensives to control fungal diseases has by consequence to impact negatively over the environment and human health, this way, the use of plant extracts with antifungal properties along with proper cultural management makes viable an alternative plant production control, specially for familiar and organic cultures. The objective of this study was to perform phytochemical and antioxidant analysis of Byrsonima crassifolia (canjiqueira) barks and evaluate its antifungal potential over Fusarium solani and Sclerotinia sclerotiorum mycelial growth. The ethanol extract from plants collected in Pantanal, Mato Grosso do Sul, Brazil was submitted to phytochemical prospection, total phenol and flavonoids quantification and antioxidant activiy determination (DPPH). To evaluate antifungal activity concentrations of 800, 1200, 1600, 2000 and 2400 µg 100 mL-1 of ethanol extract were used. Which concentration was separately incorporated in agar (PDA) and shed in Petri dishes, followed by the fungi mycelial disc where the colonies diameter was measured daily. Negatives control with agar without extract and agar with an ethanol solution were used. The B. crassifolia ethanol extract presented inhibitory activity over the fungi studied where concentrations of 800 and 1600 µg 100 mL-1, inhibited 38% of the mycelial growth of F. solani; to S. sclerotiorum the best concentration was 2400 µg 100 mL1, reducing 37.5%. The antifungal bark extract potential of this specie is attributed to phenolic compounds and to triterpenes derivatives.

  15. Characterization of volatile constituents of Haplopappus greenei and studies on the antifungal activity against phytopathogens.

    Science.gov (United States)

    Demirci, Betul; Baser, Kemal Husnu Can; Tabanca, Nurhayat; Wedge, David E

    2006-04-19

    Essential oil of Haplopappus greenei A. Gray was obtained by hydrodistillation of aerial parts, which were subsequently analyzed by gas chromatography and gas chromatography-mass spectrometry. Major components were identified as carvacrol (8.7%), beta-pinene (7.6%), trans-pinocarveol (6.2%), and caryophyllene oxide (5.8%), respectively. In total, 104 components representing 84.9% of the investigated essential oil were characterized. Furthermore, the essential oil was evaluated for antimalarial, antimicrobial, and antifungal activities. However, only antifungal activity was observed against the strawberry anthracnose-causing fungal plant pathogens Colletotrichum acutatum, Colletotrichum fragariae, and Colletotrichum gloeosporioides using the direct overlay bioautography assay. Major essential oil components were also evaluated for antifungal activity; the carvacrol standard demonstrated nonselective activity against the three Colletotrichum species and the other compounds were inactive.

  16. Association between virulence and triazole tolerance in the phytopathogenic fungus Mycosphaerella graminicola.

    Directory of Open Access Journals (Sweden)

    Lina Yang

    Full Text Available Host resistance and synthetic antimicrobials such as fungicides are two of the main approaches used to control plant diseases in conventional agriculture. Although pathogens often evolve to overcome host resistance and antimicrobials, the majority of reports have involved qualitative host - pathogen interactions or antimicrobials targeting a single pathogen protein or metabolic pathway. Studies that consider jointly the evolution of virulence, defined as the degree of damage caused to a host by parasite infection, and antimicrobial resistance are rare. Here we compared virulence and fungicide tolerance in the fungal pathogen Mycosphaerella graminicola sampled from wheat fields across three continents and found a positive correlation between virulence and tolerance to a triazole fungicide. We also found that quantitative host resistance selected for higher pathogen virulence. The possible mechanisms responsible for these observations and their consequences for sustainable disease management are discussed.

  17. IN VITRO ANTIFUNGAL ACTIVITY OF ESSENTIAL OILS ON GROWTH OF PHYTOPATHOGENIC FUNGI

    Directory of Open Access Journals (Sweden)

    Jasenka Ćosić

    2010-12-01

    Full Text Available Eleven essential oils (clove, rosemary, cinnamon leaf, sage, scots pine, neroli, peppermint, aniseed, caraway, lavander, common thyme were tested for in vitro antifungal activity on twelve plant pathogenic fungi (Fusarium graminearum, F. verticillioides, F. subglutinans, F. oxysporum, F. avenaceum, Diaporthe helianthi, Diaporthe phaseolorum var. caulivora, Phomopsis longicolla, P. viticola, Helminthosporium sativum, Colletotrichum coccodes, Thanatephorus cucumeris. The results indicated that all oils except scots pine and neroli had antifungal activity against some or all tested fungi. The best antifungal activity had common thyme, cinnamon leaf, clove and aniseed oils. When compared to control, scots pine, neroli and sage oils stimulated mycelium growth of some investigated fungi.

  18. Biosynthesis of silver nanoparticles by phytopathogen Xanthomonas oryzae pv. oryzae strain BXO8.

    Science.gov (United States)

    Narayanan, Kannan Badri; Sakthivel, Natarajan

    2013-09-28

    Extracellular biogenic synthesis of silver nanoparticles with various shapes using the rice bacterial blight bacterium Xanthomonas oryzae pv. oryzae BXO8 is reported. The synthesized silver nanoparticles were characterized by UV-Vis spectroscopy, powder X-ray diffractometry (XRD), scanning electron microscopy, energy dispersive X-ray spectrometry, and highresolution transmission electron microscopy (HR-TEM). Based on the evidence of HR-TEM, the synthesized particles were found to be spherical, with anisotropic structures such as triangles and rods, with an average size of 14.86 nm. The crystalline nature of silver nanoparticles was evident from the bright circular spots in the SAED pattern, clear lattice fringes in the high-resolution TEM images, and peaks in the XRD pattern. The FTIR spectrum showed that biomolecules containing amide and carboxylate groups are involved in the reduction and stabilization of the silver nanoparticles. Using such a biological method for the synthesis of silver nanoparticles is a simple, viable, cost-effective, and environmentally friendly process, which can be used in antimicrobial therapy.

  19. 2-Amino-3-(Oxirane-2,3-Dicarboxamido)-Propanoyl-Valine, an Effective Peptide Antibiotic from the Epiphyte Pantoea agglomerans 48b/90 ▿

    Science.gov (United States)

    Sammer, Ulrike F.; Völksch, Beate; Möllmann, Ute; Schmidtke, Michaela; Spiteller, Peter; Spiteller, Michael; Spiteller, Dieter

    2009-01-01

    The epiphyte Pantoea agglomerans 48b/90, which has been isolated from soybean leaves, belongs to the Enterobacteriaceae, as does the plant pathogen Erwinia amylovora, which causes fire blight on rosaceous plants such as apples and leads to severe economic losses. Since P. agglomerans efficiently antagonizes phytopathogenic bacteria, the P. agglomerans strain C9-1 is used as a biocontrol agent (BlightBan C9-1). Here we describe the bioassay-guided isolation of a peptide antibiotic that is highly active against the plant pathogen E. amylovora and pathovars of Pseudomonas syringae, and we elucidate its structure. Bioassay-guided fractionation using anion-exchange chromatography followed by hydrophobic interaction liquid chromatography yielded the bioactive, highly polar antibiotic. The compound was identified as 2-amino-3-(oxirane-2,3-dicarboxamido)-propanoyl-valine by using high-resolution electrospray ionization mass spectrometry and nuclear magnetic resonance techniques. This peptide was found to be produced by three of the nine P. agglomerans strains analyzed. Notably, the biocontrol strain P. agglomerans C9-1 also produces 2-amino-3-(oxirane-2,3-dicarboxamido)-propanoyl-valine. Previously, 2-amino-3-(oxirane-2,3-dicarboxamido)-propanoyl-valine has been characterized only from Serratia plymuthica. 2-Amino-3-(oxirane-2,3-dicarboxamido)-propanoyl-valine has been shown to inhibit the growth of the human pathogen Candida albicans efficiently, but its involvement in the defense of epiphytes against phytopathogenic bacteria has not been investigated so far. PMID:19820144

  20. Diversity of culturable chitinolytic bacteria from rhizospheres of agronomic plants in Japan.

    Science.gov (United States)

    Someya, Nobutaka; Ikeda, Seishi; Morohoshi, Tomohiro; Noguchi Tsujimoto, Masako; Yoshida, Takanobu; Sawada, Hiroyuki; Ikeda, Tsukasa; Tsuchiya, Kenichi

    2011-01-01

    A total of 100 isolates of chitinolytic bacteria were obtained from the rhizospheres of various agronomic plants, and the 16S rRNA gene sequences of these isolates were determined. Phylogenetic analyses revealed that 81 isolates belonged to the classes Betaproteobacteria (39 isolates) and Gammaproteobacteria (42 isolates). Of the remaining 19 isolates, 16 belonged to the phylum Firmicutes. Clustering analysis identified 6 and 3 operational taxonomic units (OTUs) in Gammaproteobacteria and Betaproteobacteria, respectively, at the genus level. The majority of chitinolytic bacteria in Gammaproteobacteria belonged to the genera Serratia, Stenotrophomonas, and Lysobacter (14, 15, and 7 isolates, respectively) while those in Betaproteobacteria belonged to the genus Mitsuaria (37 isolates). The 16 isolates placed in Firmicutes belonged to 2 genera, Paenibacillus and Bacillus (8 isolates each). The isolates in the remaining OTUs belonged to the genera Erwinia, Aeromonas, Pseudomonas, Achromobacter, Flavobacterium, and Microbacterium, in less abundance. These results showed a wide distribution of culturable chitinolytic bacteria in the rhizospheres of various agronomic plants. Considering the potential antagonistic activity of chitinolytic enzymes against phytopathogenic fungi, which is exhibited by fungal cell wall degradation, the above-mentioned native chitinolytic bacteria in rhizospheres could potentially be utilized for the biological control of soil-borne phytopathogenic fungi.

  1. 2-amino-3-(oxirane-2,3-dicarboxamido)-propanoyl-valine, an effective peptide antibiotic from the epiphyte Pantoea agglomerans 48b/90.

    Science.gov (United States)

    Sammer, Ulrike F; Völksch, Beate; Möllmann, Ute; Schmidtke, Michaela; Spiteller, Peter; Spiteller, Michael; Spiteller, Dieter

    2009-12-01

    The epiphyte Pantoea agglomerans 48b/90, which has been isolated from soybean leaves, belongs to the Enterobacteriaceae, as does the plant pathogen Erwinia amylovora, which causes fire blight on rosaceous plants such as apples and leads to severe economic losses. Since P. agglomerans efficiently antagonizes phytopathogenic bacteria, the P. agglomerans strain C9-1 is used as a biocontrol agent (BlightBan C9-1). Here we describe the bioassay-guided isolation of a peptide antibiotic that is highly active against the plant pathogen E. amylovora and pathovars of Pseudomonas syringae, and we elucidate its structure. Bioassay-guided fractionation using anion-exchange chromatography followed by hydrophobic interaction liquid chromatography yielded the bioactive, highly polar antibiotic. The compound was identified as 2-amino-3-(oxirane-2,3-dicarboxamido)-propanoyl-valine by using high-resolution electrospray ionization mass spectrometry and nuclear magnetic resonance techniques. This peptide was found to be produced by three of the nine P. agglomerans strains analyzed. Notably, the biocontrol strain P. agglomerans C9-1 also produces 2-amino-3-(oxirane-2,3-dicarboxamido)-propanoyl-valine. Previously, 2-amino-3-(oxirane-2,3-dicarboxamido)-propanoyl-valine has been characterized only from Serratia plymuthica. 2-Amino-3-(oxirane-2,3-dicarboxamido)-propanoyl-valine has been shown to inhibit the growth of the human pathogen Candida albicans efficiently, but its involvement in the defense of epiphytes against phytopathogenic bacteria has not been investigated so far.

  2. Antimicrobial activity of glycosidase inhibitory protein isolated from Cyphomandra betacea Sendt. fruit.

    Science.gov (United States)

    Ordóñez, Roxana M; Ordóñez, Adriana A L; Sayago, Jorge E; Nieva Moreno, María I; Isla, María I

    2006-06-01

    Broad-spectrum antimicrobial activity of an invertase inhibitory protein (IIP) isolated from Cyphomandra betacea ripe fruits is documented. Minimal inhibitory concentration (MIC) values were determined by agar macrodilution and broth microdilution assays. This IIP inhibited the growth of xylophagous and phytopatogenic fungi (Ganoderma applanatum, Schizophyllum commune, Lenzites elegans, Pycnoporus sanguineous, Penicillium notatum, Aspergillus niger, Phomopsis sojae and Fusarium mango) and phytopathogenic bacteria (Xanthomonas campestris pvar vesicatoria CECT 792, Pseudomonas solanacearum CECT 125, Pseudomonas corrugata CECT 124, Pseudomonas syringae pv. syringae and Erwinia carotovora var carotovora). The IIP concentration required to completely inhibit the growth of all studied fungi ranged from 7.8 to 62.5 microg/ml. Phytopatogenic bacteria were the most sensitive, with MIC values between 7.8 and 31.25 microg/ml. Antifungal and antibacterial activities can be associated with their ability to inhibit hydrolytic enzymes. Our results indicate the possible participation of IIP in the plant defense mechanism and its potential application as a biocontrol agent against phytopathogenic fungi and bacteria.

  3. 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...

  4. First report of latent infection of Cyperus rotundus caused by a biovar 3 Dickeya sp. (Syn. Erwinia chrysanthemi) in Israel

    NARCIS (Netherlands)

    Tsror, L.; Lebiush, S.; Erlich, O.; Ben-Daniel, B.; Wolf, van der J.M.

    2010-01-01

    Recent outbreaks of potato blackleg in Israel, caused by Dickeya spp., on plants grown from seed tubers imported from Northern Europe, are of a great concern. The warm climatic conditions during the growing season favour disease expression, and may result in the establishment of the pathogen in the

  5. Assessment of recent outbreaks of Dickeya sp (syn. Erwinia chrysanthemi) slow wilt in potato crops in Israel

    NARCIS (Netherlands)

    Tsror, L.; Erlich, O.; Lebiush, S.; Hazanovsky, M.; Zig, U.; Slawiak, M.; Grabe, G.; Wolf, van der J.M.; Haar, van de J.J.

    2009-01-01

    Suspected Dickeya sp. strains were obtained from potato plants and tubers collected from commercial plots. The disease was observed on crops of various cultivars grown from seed tubers imported from the Netherlands during the spring seasons of 2004-2006, with disease incidence of 2-30% ( 10% in

  6. Assessment of recent outbreaks of Dickeya sp (syn. Erwinia chrysanthemi) slow wilt in potato crops in Israel

    OpenAIRE

    Tsror, L.; Erlich, O.; Lebiush, S.; Hazanovsky, M.; Zig, U.; Slawiak, M.; Grabe, G.; Wolf, van der, J.M.; Haar, van de, J.J.

    2009-01-01

    Suspected Dickeya sp. strains were obtained from potato plants and tubers collected from commercial plots. The disease was observed on crops of various cultivars grown from seed tubers imported from the Netherlands during the spring seasons of 2004-2006, with disease incidence of 2-30% ( 10% in average). In addition to typical wilting symptoms on the foliage, in cases of severe infection, progeny tubers were rotten in the soil. Six strains were characterised by biochemical, serological and PC...

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

    OpenAIRE

    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 Candid...

  8. Evidence that antibiotic of Pantoea agglomerans E325 is produced and active against Erwinia amylovora on stigmas of pomaceous blossoms

    Science.gov (United States)

    Pantoea agglomerans strain E325, the active ingredient in a commercial product for fire blight, previously was shown to produce a unique pH-sensitive inhibitor in vitro that is specific to E. amylovora. To evaluate antibiosis as a mode of antagonism of E325, Tn5 mutagenesis was used to generate...

  9. The Association of Western Flower Thrips, Frankliniella occidentalis, with a near Erwinia Species Gut Bacteria: Transient of Permanent?

    NARCIS (Netherlands)

    de Vries, E.J.; Breeuwer, J.A.J.; Jacobs, G.; Mollema, G.

    2001-01-01

    Associations between insects and gut bacteria are ubiquitous, it is possible to make a distinction between permanent associations (called symbiosis), in which the same type of bacteria is present in more than one generation of the insect, and transient associations. Transient bacteria are ingested

  10. Public Health Risks of Enterobacterial Isolates Producing Extended-Spectrum β-Lactamases or AmpC β-Lactamases in Food and Food-Producing Animals: An EU Perspective of Epidemiology, Analytical Methods, Risk Factors, and Control Options

    DEFF Research Database (Denmark)

    Liebana, Ernesto; Carattoli, Alessandra; Coque, Teresa M.

    2013-01-01

    The blaESBL and blaAmpC genes are spread by plasmid-mediated integrons, insertion sequences, and transposons, some of which are homologous in food animals and humans. Cephalosporin usage in animal production is an important risk factor; restricting such use would be an effective control option....

  11. Biosynthesis of Enterobacterial Common Antigen: The ECA-Trace Phenotype of Salmonella Typhimurium and The Role of the rfe Gene in 08 Side-Chain Synthesis in Escherichia Coli

    Science.gov (United States)

    1993-03-18

    evaluation of sugar utilization or for identification purposes include MacConkey agar, triple sugar iron (TS!) agar, tetrazolium (TZ) medium, and brom...thymol blue (BTB) medium. TSI agar (Difco) was rehydrated, distributed to tubes and sterilized according to manufacturers instructions. MacConkey ...For MacConkey , rz, and BTB media, tbe appropriate sterile sugar solution was added after autoclaving to a final concentration of 1" ( MacConkey and rz

  12. Sequence diversity in the Dickeya fliC gene: phylogeny of the Dickeya genus and TaqMan® PCR for 'D. solani', new biovar 3 variant on potato in Europe.

    Science.gov (United States)

    Van Vaerenbergh, Johan; Baeyen, Steve; De Vos, Paul; Maes, Martine

    2012-01-01

    Worldwide, Dickeya (formerly Erwinia chrysanthemi) is causing soft rot diseases on a large diversity of crops and ornamental plants. Strains affecting potato are mainly found in D. dadantii, D. dianthicola and D. zeae, which appear to have a marked geographical distribution. Furthermore, a few Dickeya isolates from potato are attributed to D. chrysanthemi and D. dieffenbachiae. In Europe, isolates of Erwinia chrysanthemi biovar 1 and biovar 7 from potato are now classified in D. dianthicola. However, in the past few years, a new Dickeya biovar 3 variant, tentatively named 'Dickeya solani', has emerged as a common major threat, in particular in seed potatoes. Sequences of a fliC gene fragment were used to generate a phylogeny of Dickeya reference strains from culture collections and with this reference backbone, to classify pectinolytic isolates, i.e. Dickeya spp. from potato and ornamental plants. The reference strains of the currently recognized Dickeya species and 'D. solani' were unambiguously delineated in the fliC phylogram. D. dadantii, D. dianthicola and 'D. solani' displayed unbranched clades, while D. chrysanthemi, D. zeae and D. dieffenbachiae branched into subclades and lineages. Moreover, Dickeya isolates from diagnostic samples, in particular biovar 3 isolates from greenhouse ornamentals, formed several new lineages. Most of these isolates were positioned between the clade of 'D. solani' and D. dadantii as transition variants. New lineages also appeared in D. dieffenbachiae and in D. zeae. The strains and isolates of D. dianthicola and 'D. solani' were differentiated by a fliC sequence useful for barcode identification. A fliC TaqMan®real-time PCR was developed for 'D. solani' and the assay was provisionally evaluated in direct analysis of diagnostic potato samples. This molecular tool can support the efforts to control this particular phytopathogen in seed potato certification.

  13. Sequence diversity in the Dickeya fliC gene: phylogeny of the Dickeya genus and TaqMan® PCR for 'D. solani', new biovar 3 variant on potato in Europe.

    Directory of Open Access Journals (Sweden)

    Johan Van Vaerenbergh

    Full Text Available Worldwide, Dickeya (formerly Erwinia chrysanthemi is causing soft rot diseases on a large diversity of crops and ornamental plants. Strains affecting potato are mainly found in D. dadantii, D. dianthicola and D. zeae, which appear to have a marked geographical distribution. Furthermore, a few Dickeya isolates from potato are attributed to D. chrysanthemi and D. dieffenbachiae. In Europe, isolates of Erwinia chrysanthemi biovar 1 and biovar 7 from potato are now classified in D. dianthicola. However, in the past few years, a new Dickeya biovar 3 variant, tentatively named 'Dickeya solani', has emerged as a common major threat, in particular in seed potatoes. Sequences of a fliC gene fragment were used to generate a phylogeny of Dickeya reference strains from culture collections and with this reference backbone, to classify pectinolytic isolates, i.e. Dickeya spp. from potato and ornamental plants. The reference strains of the currently recognized Dickeya species and 'D. solani' were unambiguously delineated in the fliC phylogram. D. dadantii, D. dianthicola and 'D. solani' displayed unbranched clades, while D. chrysanthemi, D. zeae and D. dieffenbachiae branched into subclades and lineages. Moreover, Dickeya isolates from diagnostic samples, in particular biovar 3 isolates from greenhouse ornamentals, formed several new lineages. Most of these isolates were positioned between the clade of 'D. solani' and D. dadantii as transition variants. New lineages also appeared in D. dieffenbachiae and in D. zeae. The strains and isolates of D. dianthicola and 'D. solani' were differentiated by a fliC sequence useful for barcode identification. A fliC TaqMan®real-time PCR was developed for 'D. solani' and the assay was provisionally evaluated in direct analysis of diagnostic potato samples. This molecular tool can support the efforts to control this particular phytopathogen in seed potato certification.

  14. Effect of a pesticide on the extracellular slime production and pathogenicity of a non-target phytopathogen

    International Nuclear Information System (INIS)

    Balasubramanian, A.; Nilakantan, Gita

    1978-01-01

    Aldicarb (2 methyl thio) propionaldehyde-0-(methyl carbamoyl oxime), a systemic insecticide treatment altered the quantity and the quality of the extracellular polysaccharides (slime) produced by Pseudomonas solanacearum. Although 5 ppm (normal dose) aldicarb treatment reduced the quality of polysaccharides produced by the cells, the incorporation of 14 C (glucose) label and the reducing sugar contents was higher than the other treatments. Chromatographic analysis of the hydrolysed polysaccharides showed that aldicarb treatment altered their qualitative composition also. The extracellular polysaccharides produced by the pathogen treated with 5 ppm aldicarb caused wilting of tomato seedlings earlier than others, indicating thereby, that the wilt inducing factor in the slime was altered by the pesticide treatment. The limited translocation of the 14 C labelled polysaccharides in the wilted seedlings indicated mechanical blocking of the vascular system of the plants. (author)

  15. Biological Effects of a Carbohydrate-Binding Protein from an Annelid, Perinereis nuntia Against Human and Phytopathogenic Microorganisms

    OpenAIRE

    Sarkar M. A. Kawsar; Sarkar M. A. Mamun; Md S. Rahman; Hidetaro Yasumitsu; Yasuhiro Ozeki

    2010-01-01

    Lectins have a good scope in current clinical microbiology research. In the present study evaluated the antimicrobial activities of a D-galactose binding lectin (PnL) was purified from the annelid, Perinereis nuntia (polychaeta) by affinity chromatography. The molecular mass of the lectin was determined to be 32 kDa as a single polypeptide by SDS-PAGE under both reducing and non-reducing conditions. The hemagglutinating activity of the PnL showed against trypsinized and g...

  16. Recovery of a phytopathogenic bacterium Lonsdalea quercina from a lesser horseshoe bat in Moravian karst, Czech Republic

    Czech Academy of Sciences Publication Activity Database

    Kováčová, V.; Kolařík, Miroslav; Banďouchová, H.; Bartonička, T.; Berková, Hana; Havelková, B.; Hrudová, E.; Kohoutová, L.; Martínková, Natália; Zukal, Jan; Pikula, J.

    2018-01-01

    Roč. 48, č. 1 (2018), č. článku e12379. ISSN 1437-4781 R&D Projects: GA ČR(CZ) GA17-20286S Institutional support: RVO:61388971 ; RVO:68081766 Keywords : plants * Lonsdalea quercina * horseshoe bat Subject RIV: EE - Microbiology, Virology; GJ - Animal Vermins ; Diseases, Veterinary Medicine (UBO-W) OBOR OECD: Microbiology; Zoology (UBO-W) Impact factor: 1.547, year: 2016

  17. Horizontal gene transfer confers adaptive advantages to phytopathogenic fungi: a case study of catalase-peroxidase in Fusarium verticillioides

    Science.gov (United States)

    Horizontal gene transfer (HGT), the exchange and stable integration of genetic material between different evolutionary lineages, is widely observed in fungi. We hypothesize that successful stabilization of HGT elements provides adaptive advantages (e.g., virulence). Catalase/peroxidases (KatGs) are ...

  18. Opportunistic, human-pathogenic species in the Herpotrichiellaceae are phenotypically similar to saprobic or phytopathogenic species in the Venturiaceae

    NARCIS (Netherlands)

    Crous, P.W.; Schubert, K.; Braun, U.; Hoog, G.S. de; Hocking, A.D.; Shin, H.-D.; Groenewald, J.Z.

    2007-01-01

    Although morphologically similar, species of Cladophialophora (Herpotrichiellaceae) were shown to be phylogenetically distinct from Pseudocladosporium (Venturiaceae), which was revealed to be synonymous with the older genus, Fusicladium. Other than being associated with human disorders, species of

  19. Biosynthesis of a Complex Yersiniabactin-Like Natural Product via the mic Locus in Phytopathogen Ralstonia solanacearum▿†

    Science.gov (United States)

    Kreutzer, Martin F.; Kage, Hirokazu; Gebhardt, Peter; Wackler, Barbara; Saluz, Hans P.; Hoffmeister, Dirk; Nett, Markus

    2011-01-01

    A genome mining study in the plant pathogenic bacterium Ralstonia solanacearum GMI1000 unveiled a polyketide synthase/nonribosomal peptide synthetase gene cluster putatively involved in siderophore biosynthesis. Insertional mutagenesis confirmed the respective locus to be operational under iron-deficient conditions and spurred the isolation of the associated natural product. Bioinformatic analyses of the gene cluster facilitated the structural characterization of this compound, which was subsequently identified as the antimycoplasma agent micacocidin. The metal-chelating properties of micacocidin were evaluated in competition experiments, and the cellular uptake of gallium-micacocidin complexes was demonstrated in R. solanacearum GMI1000, indicating a possible siderophore role. Comparative genomics revealed a conservation of the micacocidin gene cluster in defined, but globally dispersed phylotypes of R. solanacearum. PMID:21724891

  20. Unraveling the in vitro secretome of the phytopathogen Botrytis cinerea to understand the interaction with its hosts

    Directory of Open Access Journals (Sweden)

    Raquel eGonzález-Fernández

    2015-10-01

    Full Text Available Botrytis cinerea is a necrotrophic fungus with high adaptability to different environments and hosts. It secretes a large number of extracellular proteins, which favor plant tissue penetration and colonization, thus contributing to virulence. Secretomics is a proteomics sub-discipline which study the secreted proteins and their secretion mechanisms, so-called secretome. By using proteomics as experimental approach, many secreted proteins by B. cinerea have been identified from in vitro experiments, and belonging to different functional categories: i cell wall-degrading enzymes such as pectinesterases, and endo-polygalacturonases; ii proteases involved in host protein degradation such as an aspartic protease; iii proteins related to the oxidative burst such as glyoxal oxidase; iv proteins which may induce the plant hypersensitive response such as a cerato-platanin domain-containing protein; and v proteins related to production and secretion of toxins such as malate dehydrogenase. In this mini-review, we made an overview of the proteomics contribution to the study and knowledge of the B. cinerea extracellular secreted proteins based on our current work carried out from in vitro experiments, and recent published papers both in vitro and in planta studies on this fungi. We hypothesize on the putative functions of these secreted proteins, and their connection to the biology of the B. cinerea interaction with its hosts.

  1. Defensins from the tick Ixodes scapularis are effective against phytopathogenic fungi and the human bacterial pathogen Listeria grayi

    Czech Academy of Sciences Publication Activity Database

    Tonk, Miray; Cabezas-Cruz, A.; Valdés, James J.; Rego, Ryan O. M.; Chrudimská, Tereza; Strnad, Martin; Šíma, Radek; Bell-Sakyi, L.; Franta, Z.; Vilcinskas, A.; Grubhoffer, Libor; Rahnamaeian, M.

    2014-01-01

    Roč. 7, DEC 3 2015 (2014), s. 554 ISSN 1756-3305 R&D Projects: GA ČR(CZ) GAP302/11/1901; GA MŠk(CZ) EE2.3.30.0032; GA ČR GP13-12816P Institutional support: RVO:60077344 Keywords : Antimicrobial peptide * Defensin * Listeria grayi * Fusarium spp * Ixodes scapularis * Tick cell line Subject RIV: EE - Microbiology, Virology Impact factor: 3.430, year: 2014

  2. An In Vitro Attempt for Controlling Severe Phytopathogens and Human Pathogens Using Essential Oils from Mediterranean Plants of Genus Schinus.

    Science.gov (United States)

    Elshafie, Hazem Salaheldin; Ghanney, Nadia; Mang, Stefania Mirela; Ferchichi, Ali; Camele, Ippolito

    2016-03-01

    Growing concerns about food safety and environmental protection enhanced the need for new and safe plant disease control strategies. The chemical composition of the three essential oils (EOs) extracted from leaves and fruits of Schinus terebinthifolius and leaves of Schinus molle, growing in Tunisia, was studied by GC and GC-MS. In all, 12 compounds were identified. The oils were mainly composed of terpene compounds. α-Pinene, α-phellandrene, and D-limonene were the major constituents. The aim of the current study was to evaluate the in vitro antimicrobial effectiveness of three EOs derived from plants of genus Schinus and extracted from leaves and fruits of S. terebinthifolius and leaves of S. molle. Both antifungal and antibacterial activities of the EOs were examined. The antifungal activity of the studied EOs was investigated against Colletotrichum acutatum and Botrytis cinerea in comparison with the systemic fungicide azoxystrobin used at 0.8 μL mL(-1). The antibacterial activity was evaluated against three strains of Gram-positive (G+ve) bacteria (Bacillus megaterium, Bacillus mojavensis and Clavibacter michiganensis) and four strains of Gram-negative (G-ve) bacteria (Escherichia coli, Xanthomonas campestris, Pseudomonas savastanoi, and Pseudomonas syringae pv. phaseolicola) compared with the synthetic antibiotic tetracycline at a concentration of 1600 μg mL(-1). The minimum inhibitory concentration of the studied EOs has been evaluated against the above microorganisms using the 96-well microplate method. Tested microorganisms exhibited different levels of sensitivity to each tested EO. All investigated EOs reduced the fungal mycelial growth when used at low concentrations from 250 to 1000 ppm and from 2000 to 8000 ppm against C. acutatum and B. cinerea, respectively. Higher concentrations of the same EOs exhibited a fungicidal effect against both mitosporic fungi. The EO extracted from leaves of S. terebinthifolius significantly inhibited the growth of tested bacterial strains. Nevertheless, E. coli showed a weak resistance toward the same EO and a high resistance toward the other two tested EOs. Finally, P. savastanoi and P. syringae pv. phaseolicola showed a high resistance toward all tested EOs.

  3. Ecological and phytohormonal aspects of plant volatile emission in response to single and dual infestations with herbivores and phytopathogens

    NARCIS (Netherlands)

    Ponzio, C.; Gols, R.; Pieterse, C.M.J.; Dicke, M.

    2013-01-01

    In their natural environment, plants are faced with a multitude of attackers, of which insect herbivores and plant pathogens are an important component. In response to these attacks, plants release volatile organic compounds (VOCs), which play an important role in the communication between plants

  4. Essential oils of Origanum compactum and Thymus vulgaris exert a protective effect against the phytopathogen Allorhizobium vitis.

    Science.gov (United States)

    Habbadi, Khaoula; Meyer, Thibault; Vial, Ludovic; Gaillard, Vincent; Benkirane, Rachid; Benbouazza, Abdellatif; Kerzaon, Isabelle; Achbani, El Hassan; Lavire, Céline

    2017-12-29

    Allorhizobium (Agrobacterium) vitis is a host-specific pathogenic bacterium that causes grapevine crown gall disease, affecting vine growth and production worldwide. The antibacterial activities of different aromatic plant essential oils were tested in vitro and in planta against A. vitis. Among the essential oils tested, those of Origanum compactum and Thymus vulgaris showed the most significant in vitro antibacterial activities, with a MIC of 0.156 and 0.312 mg/mL, respectively. A synergistic effect of these two essential oils (1:1) was observed and confirmed by the checkerboard test. Carvacrol (61.8%) and thymol (47.8%) are, respectively, the major compounds in the essential oils of O. compactum and T. vulgaris and they have been shown to be largely responsible for the antibacterial activities of their corresponding essential oils. Results obtained in vitro were reinforced by an in planta pathogenicity test. A mixture of O. compactum and T. vulgaris essential oils (1:1), inoculated into the injured stem of a tomato plant and a grapevine at 0.312 mg/mL as a preventive treatment, reduced both the number of plants developing gall symptoms and the size of the tumors.

  5. Role of sterol 3-ketoreductase sensitivity in susceptibility to the fungicide fenhexamid in Botrytis cinerea and other phytopathogenic fungi.

    Science.gov (United States)

    Debieu, Danièle; Bach, Jocelyne; Montesinos, Emeline; Fillinger, Sabine; Leroux, Pierre

    2013-05-01

    The narrow-spectrum fungicide fenhexamid was introduced into French vineyards in 2000 to control grey mould caused by a complex of two cryptic species: Botrytis cinerea, the predominant species sensitive to fenhexamid, and Botrytis pseudocinerea, naturally resistant. Fenhexamid was suggested to inhibit the 3-ketoreductase involved at C-4 demethylation steps during ergosterol biosynthesis, as revealed by its effects on the B. cinerea sterol profile. Resistance monitoring studies have hitherto identified two B. cinerea fenhexamid-resistant phenotypes, both resulting from mutations in the erg27 gene encoding 3-ketoreductase. The role of 3-ketoreductase sensitivity in fungal susceptibility to fenhexamid was investigated by studying sterol profiles and microsomal 3-ketoreductase in various fungal strains. Fenhexamid does inhibit B. cinerea 3-ketoreductase activity. Erg27 mutations causing amino acid substitutions in or near the transmembrane domain strongly decrease the affinity of fenhexamid for 3-ketoreductase. Fenhexamid has very low affinities for 3-ketoreductase in inherently resistant species, whether closely related to B. cinerea, like B. pseudocinerea, or more distantly related, like Nectria haematococca. erg27 mutation and erg27 polymorphism may therefore contribute to the unfavourable binding of fenhexamid to its target, 3-ketoreductase, explaining the acquisition of fenhexamid resistance in B. cinerea and the narrow spectrum of this fungicide. © 2012 Society of Chemical Industry.

  6. Pathogen-derived nitric oxide influences formation of the appressorium infection structure in the phytopathogenic fungus Blumeria graminis.

    Science.gov (United States)

    Prats, Elena; Carver, Tim L W; Mur, Luis A J

    2008-01-01

    Nitric oxide (NO) is an important signal in plant resistance to pathogens. Here we report that NO is also generated by Blumeria graminis f.sp. hordei as a pathogenesis determinant on barley. Infection by B. graminis f.sp. hordei is dependent on appressorium formation in order to penetrate the host. Using fluorescent dye diaminofluorescein-2 diacetate (DAF-2DA) and confocal laser scanning microscopy, transient NO generation was detected within the B. graminis f.sp. hordei appressorium during its maturation. To confirm that NO was indeed being measured, DAF-2DA fluorescence was suppressed using a NO scavenger and a mammalian NO synthase inhibitor. Both chemicals affected the number of appressorial lobes produced by the fungus. These data indicate that NO plays a key role in formation of B. graminis f.sp. hordei appressoria.

  7. New butyrolactone and other metabolites from the bark of Endlicheria arenosa against of the phytopathogen Colletotrichum tamarilloi.

    Science.gov (United States)

    Albarracín, Lady T; Delgado, Wilman; Cuca, Luis Enrique; Ávila, Mónica C

    2017-11-30

    In this work, screening of Lauraceae species for their antifungal activity against Collectotrichum tamarilloi was carried out and the ethanol extract derived from the bark of Endlicheria arenosa was found to be the best candidate. From the ethanolic extract of the bark of E. arenosa, the hexane and chloroform fractions were found to be active, from these five fatty acids were identified and two lactones were isolated. The most active fatty acid was the dodecanoic acid with a minimal inhibitory concentration (MIC) of 78.0 μM. The butyrolactone 3R,4R-licunolide A, it has not previously reported, and licunolide B show both the lowest MIC (55.3 μM). This is the first report of compounds of natural origin as growth inhibitors of C. tamarilloi.

  8. Transcription Factor Repertoire of Necrotrophic Fungal Phytopathogen Ascochyta rabiei: Predominance of MYB Transcription Factors As Potential Regulators of Secretome

    Directory of Open Access Journals (Sweden)

    Sandhya Verma

    2017-06-01

    Full Text Available Transcription factors (TFs are the key players in gene expression and their study is highly significant for shedding light on the molecular mechanisms and evolutionary history of organisms. During host–pathogen interaction, extensive reprogramming of gene expression facilitated by TFs is likely to occur in both host and pathogen. To date, the knowledge about TF repertoire in filamentous fungi is in infancy. The necrotrophic fungus Ascochyta rabiei, that causes destructive Ascochyta blight (AB disease of chickpea (Cicer arietinum, demands more comprehensive study for better understanding of Ascochyta-legume pathosystem. In the present study, we performed the genome-wide identification and analysis of TFs in A. rabiei. Taking advantage of A. rabiei genome sequence, we used a bioinformatic approach to predict the TF repertoire of A. rabiei. For identification and classification of A. rabiei TFs, we designed a comprehensive pipeline using a combination of BLAST and InterProScan software. A total of 381 A. rabiei TFs were predicted and divided into 32 fungal specific families of TFs. The gene structure, domain organization and phylogenetic analysis of abundant families of A. rabiei TFs were also carried out. Comparative study of A. rabiei TFs with that of other necrotrophic, biotrophic, hemibiotrophic, symbiotic, and saprotrophic fungi was performed. It suggested presence of both conserved as well as unique features among them. Moreover, cis-acting elements on promoter sequences of earlier predicted A. rabiei secretome were also identified. With the help of published A. rabiei transcriptome data, the differential expression of TF and secretory protein coding genes was analyzed. Furthermore, comprehensive expression analysis of few selected A. rabiei TFs using quantitative real-time polymerase chain reaction revealed variety of expression patterns during host colonization. These genes were expressed in at least one of the time points tested post infection. Overall, this study illustrates the first genome-wide identification and analysis of TF repertoire of A. rabiei. This work would provide the basis for further studies to dissect role of TFs in the molecular mechanisms during A. rabiei–chickpea interactions.

  9. Development of PCR-based detection methods for the quarantine phytopathogen Synchytrium endobioticum, causal agent of wart disease

    NARCIS (Netherlands)

    Boogert, van den P.H.J.F.; Gent-Pelzer, van M.P.E.; Bonants, P.J.M.; Boer, de S.H.; Wander, J.G.N.; Lévesque, C.A.; Leeuwen, van G.C.M.; Baayen, R.P.

    2005-01-01

    Abstract PCR-based methods were developed for the detection and quantification of the potato pathogen Synchytrium endobioticum in soil extracts and in planta. PCR primers, based on the internal transcribed spacer region of the multi-copy gene rDNA were tested for specificity, sensitivity and

  10. On-Site Molecular Detection of Soil-Borne Phytopathogens Using a Portable Real-Time PCR System.

    Science.gov (United States)

    DeShields, Joseph B; Bomberger, Rachel A; Woodhall, James W; Wheeler, David L; Moroz, Natalia; Johnson, Dennis A; Tanaka, Kiwamu

    2018-02-23

    On-site diagnosis of plant diseases can be a useful tool for growers for timely decisions enabling the earlier implementation of disease management strategies that reduce the impact of the disease. Presently in many diagnostic laboratories, the polymerase chain reaction (PCR), particularly real-time PCR, is considered the most sensitive and accurate method for plant pathogen detection. However, laboratory-based PCRs typically require expensive laboratory equipment and skilled personnel. In this study, soil-borne pathogens of potato are used to demonstrate the potential for on-site molecular detection. This was achieved using a rapid and simple protocol comprising of magnetic bead-based nucleic acid extraction, portable real-time PCR (fluorogenic probe-based assay). The portable real-time PCR approach compared favorably with a laboratory-based system, detecting as few as 100 copies of DNA from Spongospora subterranea. The portable real-time PCR method developed here can serve as an alternative to laboratory-based approaches and a useful on-site tool for pathogen diagnosis.

  11. Cyclic Lipopeptide Biosynthetic Genes and Products, and Inhibitory Activity of Plant-Associated Bacillus against Phytopathogenic Bacteria.

    Directory of Open Access Journals (Sweden)

    Isabel Mora

    Full Text Available The antibacterial activity against bacterial plant pathogens and its relationships with the presence of the cyclic lipopeptide (cLP biosynthetic genes ituC (iturin, bmyB (bacillomycin, fenD (fengycin and srfAA (surfactin, and their corresponding antimicrobial peptide products have been studied in a collection of 64 strains of Bacillus spp. isolated from plant environments. The most frequent antimicrobial peptide (AMP genes were bmyB, srfAA and fenD (34-50% of isolates. Most isolates (98.4% produced surfactin isoforms, 90.6% iturins and 79.7% fengycins. The antibacterial activity was very frequent and generally intense among the collection of strains because 75% of the isolates were active against at least 6 of the 8 bacterial plant pathogens tested. Hierarchical and correspondence analysis confirmed the presence of two clearly differentiated groups. One group consisted of Bacillus strains that showed a strong antibacterial activity, presented several cLPs genes and produced several isoforms of cLPs simultaneously, mainly composed of B. subtilis and B. amyloliquefaciens, although the last one was exclusive to this group. Another group was characterized by strains with very low or none antibacterial activity, that showed one or none of the cLP genes and produced a few or none of the corresponding cLPs, and was the most heterogenous group including B. subtilis, B. licheniformis, B. megaterium, B. pumilus, B. cereus and B. thuringiensis, although the last two were exclusive to this group. This work demonstrated that the antagonistic capacity of plant-associated Bacillus against plant pathogenic bacteria is related to the presence of cLP genes and to the production of the corresponding cLPs, and it is mainly associated to the species B. subtilis and B. amyloliquefaciens. Our findings would help to increase the yield and efficiency of screening methods to obtain candidate strains to biocontrol agents with a mechanism of action relaying on the production of antimicrobial cLPs.

  12. A re-evaluation of the taxonomy of phytopathogenic genera Dickeya and Pectobacterium using whole-genome sequencing data.

    Science.gov (United States)

    Zhang, Yucheng; Fan, Qiurong; Loria, Rosemary

    2016-06-01

    The genera Dickeya and Pectobacterium contain important plant pathogens. However, species from these genera are often poorly defined and some new isolates could not be assigned to any of the existing species. Due to their wide geographic distribution and lethality, a reliable and easy classification scheme for these pathogens is urgently needed. The low cost of next-generation sequencing has generated an upsurge of microbial genome sequences. Here, we present a phylogenomic and systematic analysis of the genera Dickeya and Pectobacterium. Eighty-three genomes from these two genera as well as two Brenneria genomes were included in this study. We estimated average nucleotide identity (ANI) and in silico DNA-DNA hybridization (isDDH) values in combination with the whole-genome-based phylogeny from 895 single-copy orthologous genes using these 85 genomes. Strains with ANI values of ≥96% and isDDH values of ≥70% were consistently grouped together in the phylogenetic tree. ANI, isDDH, and whole-genome-based phylogeny all support the elevation of Pectobacterium carotovorum's four subspecies (actinidiae, odoriferum, carotovorum, and brasiliense) to the species level. We also found some strains could not be assigned to any of the existing species, indicating these strains represent novel species. Furthermore, our study revealed at least ten tested genomes from these genera were misnamed in GenBank. This work highlights the potential of using whole genome sequences to re-evaluate current prokaryotic species definition and establish a unified prokaryotic species definition frame for taxonomically challenging genera. Copyright © 2016 Elsevier GmbH. All rights reserved.

  13. Revised Phylogeny and Novel Horizontally Acquired Virulence Determinants of the Model Soft Rot Phytopathogen Pectobacterium wasabiae SCC3193

    Science.gov (United States)

    Koskinen, Patrik; Nokso-Koivisto, Jussi; Pasanen, Miia; Broberg, Martin; Plyusnin, Ilja; Törönen, Petri; Holm, Liisa; Pirhonen, Minna; Palva, E. Tapio

    2012-01-01

    Soft rot disease is economically one of the most devastating bacterial diseases affecting plants worldwide. In this study, we present novel insights into the phylogeny and virulence of the soft rot model Pectobacterium sp. SCC3193, which was isolated from a diseased potato stem in Finland in the early 1980s. Genomic approaches, including proteome and genome comparisons of all sequenced soft rot bacteria, revealed that SCC3193, previously included in the species Pectobacterium carotovorum, can now be more accurately classified as Pectobacterium wasabiae. Together with the recently revised phylogeny of a few P. carotovorum strains and an increasing number of studies on P. wasabiae, our work indicates that P. wasabiae has been unnoticed but present in potato fields worldwide. A combination of genomic approaches and in planta experiments identified features that separate SCC3193 and other P. wasabiae strains from the rest of soft rot bacteria, such as the absence of a type III secretion system that contributes to virulence of other soft rot species. Experimentally established virulence determinants include the putative transcriptional regulator SirB, two partially redundant type VI secretion systems and two horizontally acquired clusters (Vic1 and Vic2), which contain predicted virulence genes. Genome comparison also revealed other interesting traits that may be related to life in planta or other specific environmental conditions. These traits include a predicted benzoic acid/salicylic acid carboxyl methyltransferase of eukaryotic origin. The novelties found in this work indicate that soft rot bacteria have a reservoir of unknown traits that may be utilized in the poorly understood latent stage in planta. The genomic approaches and the comparison of the model strain SCC3193 to other sequenced Pectobacterium strains, including the type strain of P. wasabiae, provides a solid basis for further investigation of the virulence, distribution and phylogeny of soft rot bacteria and, potentially, other bacteria as well. PMID:23133391

  14. Revised phylogeny and novel horizontally acquired virulence determinants of the model soft rot phytopathogen Pectobacterium wasabiae SCC3193.

    Directory of Open Access Journals (Sweden)

    Johanna Nykyri

    Full Text Available Soft rot disease is economically one of the most devastating bacterial diseases affecting plants worldwide. In this study, we present novel insights into the phylogeny and virulence of the soft rot model Pectobacterium sp. SCC3193, which was isolated from a diseased potato stem in Finland in the early 1980s. Genomic approaches, including proteome and genome comparisons of all sequenced soft rot bacteria, revealed that SCC3193, previously included in the species Pectobacterium carotovorum, can now be more accurately classified as Pectobacterium wasabiae. Together with the recently revised phylogeny of a few P. carotovorum strains and an increasing number of studies on P. wasabiae, our work indicates that P. wasabiae has been unnoticed but present in potato fields worldwide. A combination of genomic approaches and in planta experiments identified features that separate SCC3193 and other P. wasabiae strains from the rest of soft rot bacteria, such as the absence of a type III secretion system that contributes to virulence of other soft rot species. Experimentally established virulence determinants include the putative transcriptional regulator SirB, two partially redundant type VI secretion systems and two horizontally acquired clusters (Vic1 and Vic2, which contain predicted virulence genes. Genome comparison also revealed other interesting traits that may be related to life in planta or other specific environmental conditions. These traits include a predicted benzoic acid/salicylic acid carboxyl methyltransferase of eukaryotic origin. The novelties found in this work indicate that soft rot bacteria have a reservoir of unknown traits that may be utilized in the poorly understood latent stage in planta. The genomic approaches and the comparison of the model strain SCC3193 to other sequenced Pectobacterium strains, including the type strain of P. wasabiae, provides a solid basis for further investigation of the virulence, distribution and phylogeny of soft rot bacteria and, potentially, other bacteria as well.

  15. The antifungal activity of essential oils in combination with warm air flow against postharvest phytopathogenic fungi in apples

    Czech Academy of Sciences Publication Activity Database

    Franková, A.; Šmíd, J.; Bernardos, A.; Finkousová, A.; Maršík, Petr; Novotný, D.; Legarová, V.; Půlkrábek, J.; Klouček, P.

    2016-01-01

    Roč. 68, OCT (2016), s. 62-68 ISSN 0956-7135 R&D Projects: GA MŠk(CZ) LD13013 Institutional support: RVO:61389030 Keywords : Essential oil vapor phase * Antifungal * Warm air flow Subject RIV: EI - Biotechnology ; Bionics Impact factor: 3.496, year: 2016

  16. Characterization of a novel double-stranded RNA mycovirus conferring hypovirulence from the phytopathogenic fungus Botryosphaeria dothidea.

    Science.gov (United States)

    Zhai, Lifeng; Xiang, Jun; Zhang, Meixin; Fu, Min; Yang, Zuokun; Hong, Ni; Wang, Guoping

    2016-06-01

    A novel double-stranded RNA (dsRNA) virus, designated as Botryosphaeria dothidea RNA virus 1 (BdRV1), isolated from a hypovirulent strain YZN115 of Botryosphaeria dothidea was biologically and molecularly characterized. The genome of BdRV1 comprises of five dsRNAs. Each dsRNA contains a single open reading frame. The proteins encoded by dsRNA1-4 shared significant amino acid identities of 55%, 47%, 43% and 53% with the corresponding proteins of Aspergillus fumigatus tetramycovirus-1. DsRNA1, 3, and 4 of BdRV1 encoded an RNA-dependent RNA polymerase, a viral methyltransferase, and a P-A-S-rich protein, respectively. Function of proteins encoded by the dsRNA2 and dsRNA5 were unknown. BdRV1 conferred hypovirulence for its host and could be transmitted through conidia and hyphae contact. Copyright © 2016 Elsevier Inc. All rights reserved.

  17. Production of antioomycete compounds active against the phytopathogens Phytophthora sojae and Aphanomyces cochlioides by clavicipitoid entomopathogenic fungi.

    Science.gov (United States)

    Putri, Sastia Prama; Ishido, Kei-Ichi; Kinoshita, Hiroshi; Kitani, Shigeru; Ihara, Fumio; Sakihama, Yasuko; Igarashi, Yasuhiro; Nihira, Takuya

    2014-05-01

    A total of 412 strains belonging to 14 genera of clavicipitoid entomopathogenic fungi (EPF) were screened for activities against two economically important plant pathogenic oomycetes, Phytophthora sojae and Aphanomyces cochlioides. To identify the antioomycete compounds produced by EPF, the extracts of 13 highly active EPF strains were characterized in detail by high performance liquid chromatography with diode array detection and high-resolution mass spectrometric detection and antioomycete assay. The antioomycete activity of several Metarhizium extracts was associated with previously isolated aurovertins, fungerin, N-(methyl-3-oxodec-6-enoyl)-2-pyrroline, and N-(methyl-3-oxodecanoyl)-2-pyrroline. The depsipeptide beauvericin was confirmed to be one of the active principles of three strains of Isaria tenuipes, which strongly inhibited mycelial growth of both P. sojae and A. cochlioides. Two known bioactive metabolites, paecilosetin and aranorosinol A, together with a novel and potent antioomycete compound, farinomalein, were isolated from the extracts of Isaria farinosa and all compounds were confirmed to have antioomycete activity. Identification of 8 antioomycete compounds from 13 clavicipitioid EPF demonstrated a new potential use of EPF as a source of compounds for the control of soil-borne plant pathogenic oomycetes. Copyright © 2013 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

  18. Tolerance to oxidative stress is required for maximal xylem colonization by the xylem-limited bacterial phytopathogen, Xylella fastidiosa.

    Science.gov (United States)

    Wang, Peng; Lee, Yunho; Igo, Michele M; Roper, M Caroline

    2017-09-01

    Bacterial plant pathogens often encounter reactive oxygen species (ROS) during host invasion. In foliar bacterial pathogens, multiple regulatory proteins are involved in the sensing of oxidative stress and the activation of the expression of antioxidant genes. However, it is unclear whether xylem-limited bacteria, such as Xylella fastidiosa, experience oxidative stress during the colonization of plants. Examination of the X. fastidiosa genome uncovered only one homologue of oxidative stress regulatory proteins, OxyR. Here, a knockout mutation in the X. fastidiosa oxyR gene was constructed; the resulting strain was significantly more sensitive to hydrogen peroxide (H 2 O 2 ) relative to the wild-type. In addition, during early stages of grapevine infection, the survival rate was 1000-fold lower for the oxyR mutant than for the wild-type. This supports the hypothesis that grapevine xylem represents an oxidative environment and that X. fastidiosa must overcome this challenge to achieve maximal xylem colonization. Finally, the oxyR mutant exhibited reduced surface attachment and cell-cell aggregation and was defective in biofilm maturation, suggesting that ROS could be a potential environmental cue stimulating biofilm development during the early stages of host colonization. © 2016 BSPP AND JOHN WILEY & SONS LTD.

  19. Crystallization, data collection and phasing of the molybdate-binding protein of the phytopathogen Xanthomonas axonopodis pv. citri

    International Nuclear Information System (INIS)

    Santacruz, C. P.; Balan, A.; Ferreira, L. C. S.; Barbosa, J. A. R. G.

    2006-01-01

    The molybdate-binding protein (ModA) from X. axonopodis pv. citri was crystallized with sodium molybdate in the presence of PEG or sulfate. The crystal diffracted to a maximum resolution of 1.7 Å and belongs to the orthorhombic space group C222 1, with unit-cell parameters a = 68.15, b = 172.14, c = 112.04 Å. Xanthomonas axonopodis pv. citri ModA protein is the ABC periplasmic binding component responsible for the capture of molybdate. The protein was crystallized with sodium molybdate using the hanging-drop vapour-diffusion method in the presence of PEG or sulfate. X-ray diffraction data were collected to a maximum resolution of 1.7 Å using synchrotron radiation. The crystal belongs to the orthorhombic space group C222 1 , with unit-cell parameters a = 68.15, b = 172.14, c = 112.04 Å. The crystal structure was solved by molecular-replacement methods and structure refinement is in progress

  20. Crystallization, data collection and phasing of the molybdate-binding protein of the phytopathogen Xanthomonas axonopodis pv. citri

    Energy Technology Data Exchange (ETDEWEB)

    Santacruz, C. P.; Balan, A.; Ferreira, L. C. S. [Departamento de Microbiologia, Instituto de Ciências Biomédicas II, Universidade de São Paulo, São Paulo, SP (Brazil); Barbosa, J. A. R. G., E-mail: joao@lnls.br [Centro de Biologia Molecular e Estrutural (CeBiMe), Laboratório Nacional de Luz Síncrotron (LNLS), CP 6192, Campinas, SP 13084-971 (Brazil); Departamento de Microbiologia, Instituto de Ciências Biomédicas II, Universidade de São Paulo, São Paulo, SP (Brazil)

    2006-03-01

    The molybdate-binding protein (ModA) from X. axonopodis pv. citri was crystallized with sodium molybdate in the presence of PEG or sulfate. The crystal diffracted to a maximum resolution of 1.7 Å and belongs to the orthorhombic space group C222{sub 1,} with unit-cell parameters a = 68.15, b = 172.14, c = 112.04 Å. Xanthomonas axonopodis pv. citri ModA protein is the ABC periplasmic binding component responsible for the capture of molybdate. The protein was crystallized with sodium molybdate using the hanging-drop vapour-diffusion method in the presence of PEG or sulfate. X-ray diffraction data were collected to a maximum resolution of 1.7 Å using synchrotron radiation. The crystal belongs to the orthorhombic space group C222{sub 1}, with unit-cell parameters a = 68.15, b = 172.14, c = 112.04 Å. The crystal structure was solved by molecular-replacement methods and structure refinement is in progress.

  1. Mechanisms of Bactericide Resistance in Phytopathogenic Bacteria(Abstracts of the Research by the Winners of the Young Scientist Award)

    OpenAIRE

    Masami, NAKAJIMA; School of Agriculture, Ibaraki University

    2002-01-01

    Bactericides containing copper and streptomycin have been widely used to control bacterial plant diseases. However, the efficacy of copper and streptomycin have been reduced by the development of copper- and streptomycin-resistant bacterial strains. Understanding the mechanism of resistance is necessary for the prevention and management of resistance. In this study, the mechanisms of copper and streptomycin resistance in Pseudomonas syringae pv. actinidiae were analyzed.

  2. Presence and antimicrobial profile of gram-negative facultative anaerobe rods in patients with chronic periodontitis and gingivitis.

    Science.gov (United States)

    Gamboa, Fredy; García, Dabeiba-Adriana; Acosta, Adriana; Mizrahi, Deborah; Paz, Andreína; Martínez, Diana; Arévalo, Azucena; Aristizabal, Fabio; Abba, Martín

    2013-01-01

    Chronic periodontitis is a multifactorial infectious disease associated with Gram-negative anaerobes which are part of the subgingival microflora. In recent years, studies have been conducted to assess the presence of Gram-negative facultative anaerobes (Enterobacteriaceae) and their participation in the development and progression of chronic periodontitis. The aim of this study was to determine the presence of Enterobacteriaceae in patients with chronic periodontitis and gingivitis and to assess antimicrobial susceptibility of clinical isolates. A descriptive, observational study was performed including 64 patients with chronic periodontitis and 22 patients with gingivitis. Microbiological samples were taken from the gingival sulcus using paper points, which then were placed in thioglycollate broth. Samples were incubated for 4 hours at 37 degrees C and finally replated on MacConkey agar Bacteria were identified using the API-20E system (Biomerieux, France) and antimicrobial susceptibility was determined using the disk diffusion method. The evaluation of samples showed presence of 29 enterobacterial species distributed as follows: 7 in the group with gingivitis and 22 in the group with chronic periodontitis. In the chronic periodontitis group the most common species were: K. oxytoca n = 5, S. liquefaciens n = 4 and K. pneumoniae and E. coli with n = 3. The gingivitis group had the highest frequency of Erwinia sp. (n = 2). Clinical isolates showed very low sensitivity levels to beta-lactam ampicillin and amoxicillin/ clavulanic acid, 17.2% and 27.6% respectively, and higher sensitivity levels to ciprofloxacin (96.6%), amikacin (79.3%), gentamicin (68.9%) and ceftazidime, ceftriaxone, kanamycin and trimethoprimsulfa (65.5%). In conclusion, the existence of a high frequency of enterobacteria in patients with chronic periodontitis and gingivitis shows that periodontologists should pay greater attention to prevention protocols, and develop mechanical and antimicrobial

  3. Description of a new disease on Erythrina sp. in Martinique (French West Indies) and preliminary characterization of the causal agent as a novel Erwinia species

    OpenAIRE

    Sutra, Lydie; Prior, P.; Perlemoine, K.; Risede,; Cao-Van, P.; Gardan, Louis

    1999-01-01

    In spring 1995, symptoms of partial defoliation were observed an Erythrina indica var. fastigiata trees, commonly used as windbreaks in banana plantations in Martinique (French West Indies). Browning of the bark surface was consistently observed at the base of defoliated branches. Bacteria were isolated as nearly pure cultures from typical necrotic lesions an the bark. Results of Gram stain, staining of flagella and biochemical tests indicated that all isolates belonged to the Enterobacteriac...

  4. Scavenging Iron: A Novel Mechanism of Plant Immunity Activation by Microbial Siderophores1[C][W

    Science.gov (United States)

    Aznar, Aude; Chen, Nicolas W.G.; Rigault, Martine; Riache, Nassima; Joseph, Delphine; Desmaële, Didier; Mouille, Grégory; Boutet, Stéphanie; Soubigou-Taconnat, Ludivine; Renou, Jean-Pierre; Thomine, Sébastien; Expert, Dominique; Dellagi, Alia

    2014-01-01

    Siderophores are specific ferric iron chelators synthesized by virtually all microorganisms in response to iron deficiency. We have previously shown that they promote infection by the phytopathogenic enterobacteria Dickeya dadantii and Erwinia amylovora. Siderophores also have the ability to activate plant immunity. We have used complete Arabidopsis transcriptome microarrays to investigate the global transcriptional modifications in roots and leaves of Arabidopsis (Arabidopsis thaliana) plants after leaf treatment with the siderophore deferrioxamine (DFO). Physiological relevance of these transcriptional modifications was validated experimentally. Immunity and heavy-metal homeostasis were the major processes affected by DFO. These two physiological responses could be activated by a synthetic iron chelator ethylenediamine-di(o-hydroxyphenylacetic) acid, indicating that siderophores eliciting activities rely on their strong iron-chelating capacity. DFO was able to protect Arabidopsis against the pathogenic bacterium Pseudomonas syringae pv tomato DC3000. Siderophore treatment caused local modifications of iron distribution in leaf cells visible by ferrocyanide and diaminobenzidine-H2O2 staining. Metal quantifications showed that DFO causes a transient iron and zinc uptake at the root level, which is presumably mediated by the metal transporter iron regulated transporter1 (IRT1). Defense gene expression and callose deposition in response to DFO were compromised in an irt1 mutant. Consistently, plant susceptibility to D. dadantii was increased in the irt1 mutant. Our work shows that iron scavenging is a unique mechanism of immunity activation in plants. It highlights the strong relationship between heavy-metal homeostasis and immunity. PMID:24501001

  5. Champacyclin, a New Cyclic Octapeptide from Streptomyces Strain C42 Isolated from the Baltic Sea

    Directory of Open Access Journals (Sweden)

    Alexander Pesic

    2013-12-01

    Full Text Available New isolates of Streptomyces champavatii were isolated from marine sediments of the Gotland Deep (Baltic Sea, from the Urania Basin (Eastern Mediterranean, and from the Kiel Bight (Baltic Sea. The isolates produced several oligopeptidic secondary metabolites, including the new octapeptide champacyclin (1a present in all three strains. Herein, we report on the isolation, structure elucidation and determination of the absolute stereochemistry of this isoleucine/leucine (Ile/Leu = Xle rich cyclic octapeptide champacyclin (1a. As 2D nuclear magnetic resonance (NMR spectroscopy could not fully resolve the structure of (1a, additional information on sequence and configuration of stereocenters were obtained by a combination of multi stage mass spectrometry (MSn studies, amino acid analysis, partial hydrolysis and subsequent enantiomer analytics with gas chromatography positive chmical ionization/electron impact mass spectrometry (GC-PCI/EI-MS supported by comparison to reference dipeptides. Proof of the head-to-tail cyclization of (1a was accomplished by solid phase peptide synthesis (SPPS compared to an alternatively side chain cyclized derivative (2. Champacyclin (1a is likely synthesized by a non-ribosomal peptide synthetase (NRPS, because of its high content of (d-amino acids. The compound (1a showed antimicrobial activity against the phytopathogen Erwinia amylovora causing the fire blight disease of certain plants.

  6. Two novel antimicrobial defensins from rice identified by gene coexpression network analyses.

    Science.gov (United States)

    Tantong, Supaluk; Pringsulaka, Onanong; Weerawanich, Kamonwan; Meeprasert, Arthitaya; Rungrotmongkol, Thanyada; Sarnthima, Rakrudee; Roytrakul, Sittiruk; Sirikantaramas, Supaart

    2016-10-01

    Defensins form an antimicrobial peptides (AMP) family, and have been widely studied in various plants because of their considerable inhibitory functions. However, their roles in rice (Oryza sativa L.) have not been characterized, even though rice is one of the most important staple crops that is susceptible to damaging infections. Additionally, a previous study identified 598 rice genes encoding cysteine-rich peptides, suggesting there are several uncharacterized AMPs in rice. We performed in silico gene expression and coexpression network analyses of all genes encoding defensin and defensin-like peptides, and determined that OsDEF7 and OsDEF8 are coexpressed with pathogen-responsive genes. Recombinant OsDEF7 and OsDEF8 could form homodimers. They inhibited the growth of the bacteria Xanthomonas oryzae pv. oryzae, X. oryzae pv. oryzicola, and Erwinia carotovora subsp. atroseptica with minimum inhibitory concentration (MIC) ranging from 0.6 to 63μg/mL. However, these OsDEFs are weakly active against the phytopathogenic fungi Helminthosporium oryzae and Fusarium oxysporum f.sp. cubense. This study describes a useful method for identifying potential plant AMPs with biological activities. Copyright © 2016 Elsevier Inc. All rights reserved.

  7. Antiproliferative, antifungal, and antibacterial activities of endophytic alternaria species from cupressaceae.

    Science.gov (United States)

    Soltani, Jalal; Hosseyni Moghaddam, Mahdieh S

    2014-09-01

    Recent research has shown the bioprospecting of endophytic fungi from Cupressaceae. Here, we further uncover that the healthy cypress plants such as Cupressus arizonica, Cupressus sempervirens var. cereiformis, and Thuja orientalis host highly bioactive endophytic Alternaria fungal species. Indeed, endophytic Alternaria alternata, Alternaria pellucida, and Alternaria tangelonis were recovered from healthy Cupressaceous trees. Biodiversity and bioactivity of recovered endophytic Alternaria species were a matter of biogeography and host identity. We further extracted such Alternaria's metabolites and highlighted their significant antiproliferative, growth inhibitory, and antibacterial activities against the model target fungus Pyricularia oryzae and the model pathogenic bacteria Bacillus sp., Erwinia amylovora, and Pseudomonas syringae. In vitro assays also indicated that endophytic Alternaria species significantly inhibited the growth of cypress fungal phytopathogens Diplodia seriata, Phaeobotryon cupressi, and Spencermartinsia viticola. In conclusion, since the recovered Alternaria species were originally reported as pathogenic and allergenic fungi, our findings suggest a possible ecological niche for them inside the foliar tissues of Cupressaceous trees. Moreover, in this study, the significant bioactivities of endophytic Alternaria species in association with Cupressaceae plant family are reported.

  8. Phenotypic comparison of clinical and plant-beneficial strains of Pantoea agglomerans.

    Science.gov (United States)

    Bonaterra, Anna; Badosa, Esther; Rezzonico, Fabio; Duffy, Brion; Montesinos, Emilio

    2014-06-01

    Certain strains of Pantoea are used as biocontrol agents for the suppression of plant diseases. However, their commercial registration is hampered in some countries because of biosafety concerns. This study compares clinical and plant-beneficial strains of P. agglomerans and related species using a phenotypic analysis approach in which plant-beneficial effects, adverse effects in nematode models, and toxicity were evaluated. Plant-beneficial effects were determined as the inhibition of apple fruit infection by Penicillium expansum and apple flower infection by Erwinia amylovora. Clinical strains had no general inhibitory activity against infection by the fungal or bacterial plant pathogens, as only one clinical strain inhibited P. expansum and three inhibited E. amylovora. By contrast, all biocontrol strains showed activity against at least one of the phytopathogens, and three strains were active against both. The adverse effects in animals were evaluated in the plant-parasitic nematode Meloidogyne javanica and the bacterial-feeding nematode Caenorhabditis elegans. Both models indicated adverse effects of the two clinical strains but not of any of the plant-beneficial strains. Toxicity was evaluated by means of hemolytic activity in blood, and genotoxicity with the Ames test. None of the strains, whether clinical or plant-beneficial, showed any evidence of toxicity. Copyright© by the Spanish Society for Microbiology and Institute for Catalan Studies.

  9. CorA, the magnesium/nickel/cobalt transporter, affects virulence and extracellular enzyme production in the soft rot pathogen Pectobacterium carotovorum.

    Science.gov (United States)

    Kersey, Caleb M; Agyemang, Paul A; Dumenyo, C Korsi

    2012-01-01

    Pectobacterium carotovorum (formerly Erwinia carotovora ssp. carotovora) is a phytopathogenic bacterium that causes soft rot disease, characterized by water-soaked soft decay, resulting from the action of cell wall-degrading exoenzymes secreted by the pathogen. Virulence in soft rot bacteria is regulated by environmental factors, host and bacterial chemical signals, and a network of global and gene-specific bacterial regulators. We isolated a mini-Tn5 mutant of P. carotovorum that is reduced in the production of extracellular pectate lyase, protease, polygalacturonase and cellulase. The mutant is also decreased in virulence as it macerates less host tissues than its parent and is severely impaired in multiplication in planta. The inactivated gene responsible for the reduced virulent phenotype was identified as corA. CorA, a magnesium/nickel/cobalt membrane transporter, is the primary magnesium transporter for many bacteria. Compared with the parent, the CorA(-) mutant is cobalt resistant. The mutant phenotype was confirmed in parental strain P. carotovorum by marker exchange inactivation of corA. A functional corA(+) DNA from P. carotovorum restored exoenzyme production and pathogenicity to the mutants. The P. carotovorum corA(+) clone also restored motility and cobalt sensitivity to a CorA(-) mutant of Salmonella enterica. These data indicate that CorA is required for exoenzyme production and virulence in P. carotovorum. © 2011 THE AUTHORS. MOLECULAR PLANT PATHOLOGY © 2011 BSPP AND BLACKWELL PUBLISHING LTD.

  10. Quorum sensing-controlled Evr regulates a conserved cryptic pigment biosynthetic cluster and a novel phenomycin-like locus in the plant pathogen, Pectobacterium carotovorum.

    Science.gov (United States)

    Williamson, Neil R; Commander, Paul M B; Salmond, George P C

    2010-07-01

    Pectobacterium carotovorum SCRI193 is a phytopathogenic Gram-negative bacterium. In this study, we have identified a novel cryptic pigment biosynthetic locus in P. carotovorum SCRI193 which we have called the Pectobacterium orange pigment (pop) cluster. The pop cluster is flanked by two tRNA genes and contains genes that encode non-ribosomal peptide synthases and polyketide synthase and produces a negatively charged polar orange pigment. Orange pigment production is activated when an adjacent transcriptional activator sharing sequence similarity with the Erwinia virulence regulator (Evr) is overexpressed. Evr was shown to positively activate its own transcription and that of the pigment biosynthetic genes and an unlinked locus encoding a phenomycin homologue. In addition, the expression of Evr and orange pigment production was shown to be regulated by N-(3-oxohexanoyl)-HSL (OHHL) quorum sensing and have a virulence phenotype in potato. Finally, by comparative genomics and Southern blotting we demonstrate that this pigment biosynthetic cluster is present in multiple P. carotovorum spp., Pectobacterium brasiliensis 1692 and a truncated version of the cluster is present in Pectobacterium atrosepticum. The conserved nature of this cluster in P. carotovorum and P. brasiliensis suggests that the pop cluster has an important function in these broad-host-range soft rotting bacteria, which is no longer required in the narrow-host-range P. atrosepticum SCRI1043.

  11. Oorzaak Erwiniaproblemen dahlia vooral Dickeya dianthicola

    NARCIS (Netherlands)

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

    2012-01-01

    Dahlia kan, net als een aantal andere bol- en knolgewassen, worden aangetast door Erwinia. Bacterieverwelkingsziekte in dahlia wordt veroorzaakt door Erwinia chrysanthemi, een bacterie die tegenwoordig Dickeya heet. PPO Bloembollen onderzocht welke problemen Dickeya veroorzaakt bij dahlia en wat er

  12. In Vitro and In Vivo Plant Growth Promoting Activities and DNA Fingerprinting of Antagonistic Endophytic Actinomycetes Associates with Medicinal Plants.

    Science.gov (United States)

    Passari, Ajit Kumar; Mishra, Vineet Kumar; Gupta, Vijai Kumar; Yadav, Mukesh Kumar; Saikia, Ratul; Singh, Bhim Pratap

    2015-01-01

    Endophytic actinomycetes have shown unique plant growth promoting as well as antagonistic activity against fungal phytopathogens. In the present study forty-two endophytic actinomycetes recovered from medicinal plants were evaluated for their antagonistic potential and plant growth-promoting abilities. Twenty-two isolates which showed the inhibitory activity against at least one pathogen were subsequently tested for their plant-growth promoting activities and were compared genotypically using DNA based fingerprinting, including enterobacterial repetitive intergenic consensus (ERIC) and BOX repetitive elements. Genetic relatedness based on both ERIC and BOX-PCR generates specific patterns corresponding to particular genotypes. Exponentially grown antagonistic isolates were used to evaluate phosphate solubilization, siderophores, HCN, ammonia, chitinase, indole-3-acetic acid production, as well as antifungal activities. Out of 22 isolates, the amount of indole-3-acetic acid (IAA) ranging between 10-32 μg/ml was produced by 20 isolates and all isolates were positive for ammonia production ranging between 5.2 to 54 mg/ml. Among 22 isolates tested, the amount of hydroxamate-type siderophores were produced by 16 isolates ranging between 5.2 to 36.4 μg/ml, while catechols-type siderophores produced by 5 isolates ranging from 3.2 to 5.4 μg/ml. Fourteen isolates showed the solubilisation of inorganic phosphorous ranging from 3.2 to 32.6 mg/100ml. Chitinase and HCN production was shown by 19 and 15 different isolates, respectively. In addition, genes of indole acetic acid (iaaM) and chitinase (chiC) were successively amplified from 20 and 19 isolates respectively. The two potential strains Streptomyces sp. (BPSAC34) and Leifsonia xyli (BPSAC24) were tested in vivo and improved a range of growth parameters in chilli (Capsicum annuum L.) under greenhouse conditions. This study is the first published report that actinomycetes can be isolated as endophytes from within these

  13. Plasmid DNA studies in Lactobacillus plantarum strains isolated from olive fermentations: production of and immunity to plantaricin OL15 is associated to a 9.6 Kb plasmid (pOL15

    Directory of Open Access Journals (Sweden)

    Mourad, Kacem

    2007-06-01

    Full Text Available Previously 12 Lactobacillus plantarum strains were isolated from fermented olives. Among these, only L. plantarum OL15 produced bacteriocin (plantaricin OL15. In this study, the 12 strains were examined for plasmid DNA content. Of these, 9 strains have shown one to three plasmid bands ranging in size from 5.4 to 12.2 kb. L. plantarum OL15 exhibited one plasmid (9.6 kb which was named pOL15. After curing with novobiocin and ethidium bromide, the plasmid profile analysis of non producing derivatives, showed that the 9.6 kb plasmid pOL15 harbored by the parental strain had been lost in all cases and none of them regained the ability to produce plantaricin OL15 suggesting that the production of plantaricin OL15 is plasmid linked. Plantaricin OL15 was not inactived by amylase and lipase suggesting that plantaricin OL15 activity was not dependent on the presence of either a carbohydrate or lipid moiety. Plantaricin OL15 showed activity against lactic acid bacteria of different species and also against olive spoilage and phytopathogenic bacteria, including Pseudomonas and Erwinia.En un estudio previo, se aislaron 12 cepas de Lactobacillus plantarum a partir de aceitunas fermentadas. Entre ellas, solo L. plantarum OL15 produjo bacteriocinas (plantaricin OL15. En este estudio, se examinó el contenido de AND plásmido en las 12 cepas citadas. Entre ellas, 9 cepas han mostrado de una a tres bandas de plásmido con tamaños en el rango de 5.4 a 12.2 kb. L. plantarum OL15 exhibió un plásmido (9.6 kb que se denominó pOL15. Después del curado con novobiocina y bromuro de etidio, la pérdida del plásmido pOL15 asociada a la pérdida de su facultad para producir plantaricin OL15, sugiere que la producción de plantaricina OL15 está ligada al plásmido. La plantaricin OL15 no se inactivó por amilasa ni por lipasa sugiriendo que su actividad no es dependiente de la presencia de carbohidratos o lípidos. La plantaricina OL15 mostró actividad frente a

  14. Mini-Tn7 Insertion in an Artificial attTn7 Site Enables Depletion of the Essential Master Regulator CtrA in the Phytopathogen Agrobacterium tumefaciens.

    Science.gov (United States)

    Figueroa-Cuilan, Wanda; Daniel, Jeremy J; Howell, Matthew; Sulaiman, Aliyah; Brown, Pamela J B

    2016-08-15

    Mechanistic studies of many processes in Agrobacterium tumefaciens have been hampered by a lack of genetic tools for characterization of essential genes. In this study, we used a Tn7-based method for inducible control of transcription from an engineered site on the chromosome. We demonstrate that this method enables tighter control of inducible promoters than plasmid-based systems and can be used for depletion studies. The method enables the construction of depletion strains to characterize the roles of essential genes in A. tumefaciens Here, we used the strategy to deplete the alphaproteobacterial master regulator CtrA and found that depletion of this essential gene results in dramatic rounding of cells, which become nonviable. Agrobacterium tumefaciens is a bacterial plant pathogen and natural genetic engineer. Thus, studies of essential processes, including cell cycle progression, DNA replication and segregation, cell growth, and division, may provide insights for limiting disease or improving biotechnology applications. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

  15. Comparison of genetic variability between Czech and foreign isolates of phytopathogenic bacteria Clavibacter michiganensis subsp. sepedonicus by Rep-PCR technique

    Czech Academy of Sciences Publication Activity Database

    Fousek, Jan; Mráz, Ivan; Petrzik, Karel

    2002-01-01

    Roč. 47, - (2002), s. 450-454 ISSN 0015-5632 R&D Projects: GA ČR GA522/00/0887 Keywords : Clavibacter michiganensis * genetic variability Subject RIV: EE - Microbiology, Virology Impact factor: 0.979, year: 2002

  16. The Filament-specific Rep1-1 Repellent of the Phytopathogen Ustilago maydis Forms Functional Surface-active Amyloid-like Fibrils

    NARCIS (Netherlands)

    Teertstra, Wieke R.; van der Velden, Gisela J.; de Jong, Jan F.; Kruijtzer, John A. W.; Liskamp, Rob M. J.; Kroon-Batenburg, Loes M. J.; Muller, Wally H.; Gebbink, Martijn F. B. G.; Wosten, Han A. B.

    2009-01-01

    Repellents of the maize pathogen Ustilago maydis are involved in formation of hydrophobic aerial hyphae and in cellular attachment. These peptides, called Rep1-1 to Rep1-11, are encoded by the rep1 gene and result from cleavage of the precursor protein Rep1 during passage of the secretion pathway.

  17. Molecular characterization of a bipartite double-stranded RNA virus and its satellite-like RNA co-infecting the phytopathogenic fungus Sclerotinia sclerotiorum

    Directory of Open Access Journals (Sweden)

    Lijiang eLiu

    2015-05-01

    Full Text Available A variety of mycoviruses have been found in Sclerotinia sclerotiorum. In this study, we report a novel mycovirus Sclerotinia sclerotiorum botybirnavirus 1 (SsBRV1 that was originally isolated from the hypovirulent strain SCH941 of S. sclerotiorum. SsBRV1 has rigid spherical virions that are ~38 nm in diameter, and three dsRNA segments (dsRNA1, 2 and 3 with lengths of 6.4, 6.0 and 1.7 kbp, respectively were packaged in the virions. dsRNA1 encodes a cap-pol fusion protein, and dsRNA2 encodes a polyprotein with unknown functions but contributes to the formation of virus particles. The dsRNA3 is dispensable and may be a satellite-like RNA (SatlRNA of SsBRV1. Although phylogenetic analysis of the RdRp domain demonstrated that SsBRV1 is related to Botrytis porri RNA virus 1 (BpRV1 and Ustilago maydis dsRNA virus-H1 (UmV-H1, the structure proteins of SsBRV1 do not have any significant sequence similarities with other known viral proteins with the exception of those of BpRV1. SsBRV1 carrying dsRNA3 seems to have no obvious effects on the colony morphology, but can significantly reduce the growth rate and virulence of S. sclerotiorum. Notably, a growth hormone receptor binding domain (GHBP, Pfam12772 is detected in ORF2-encoded protein of SsBRV1, which have not been reported in any other viruses. These findings provide new insights into the virus taxonomy, virus evolution and the interactions between SsBRV1 and the fungal hosts.

  18. Increased metabolite production by deletion of an HDA1-type histone deacetylase in the phytopathogenic fungi, Magnaporthe oryzae (Pyricularia oryzae) and Fusarium asiaticum.

    Science.gov (United States)

    Maeda, K; Izawa, M; Nakajima, Y; Jin, Q; Hirose, T; Nakamura, T; Koshino, H; Kanamaru, K; Ohsato, S; Kamakura, T; Kobayashi, T; Yoshida, M; Kimura, M

    2017-11-01

    Histone deacetylases (HDACs) play an important role in the regulation of chromatin structure and gene expression. We found that dark pigmentation of Magnaporthe oryzae (anamorph Pyricularia oryzae) ΔMohda1, a mutant strain in which an orthologue of the yeast HDA1 was disrupted by double cross-over homologous recombination, was significantly stimulated in liquid culture. Analysis of metabolites in a ΔMohda1 mutant culture revealed that the accumulation of shunt products of the 1,8-dihydroxynaphthalene melanin and ergosterol pathways were significantly enhanced compared to the wild-type strain. Northern blot analysis of the ΔMohda1 mutant revealed transcriptional activation of three melanin genes that are dispersed throughout the genome of M. oryzae. The effect of deletion of the yeast HDA1 orthologue was also observed in Fusarium asiaticum from the Fusarium graminearum species complex; the HDF2 deletion mutant produced increased levels of nivalenol-type trichothecenes. These results suggest that histone modification via HDA1-type HDAC regulates the production of natural products in filamentous fungi. Natural products of fungi have significant impacts on human welfare, in both detrimental and beneficial ways. Although HDA1-type histone deacetylase is not essential for vegetative growth, deletion of the gene affects the expression of clustered secondary metabolite genes in some fungi. Here, we report that such phenomena are also observed in physically unlinked genes required for melanin biosynthesis in the rice blast fungus. In addition, production of Fusarium trichothecenes, previously reported to be unaffected by HDA1 deletion, was significantly upregulated in another Fusarium species. Thus, the HDA1-inactivation strategy may be regarded as a general approach for overproduction and/or discovery of fungal metabolites. © 2017 The Society for Applied Microbiology.

  19. The MAP kinase-encoding gene MgFus3 of the non-appressorium phytopathogen Mycosphaerella graminicola is required for penetration and in vitro pycnidia formation

    NARCIS (Netherlands)

    Cousin, A.; Mehrabi, R.; Guilleroux, M.; Dufresne, M.; Lee, van der T.A.J.; Waalwijk, C.; Langin, T.; Kema, G.H.J.

    2006-01-01

    In eukaryotes, a family of serine/threonine protein kinases known as mitogen-activated protein kinases (MAPKs) is involved in the transduction of a variety of extracellular signals and in the regulation of growth and development. We identified a MAPK-encoding gene in Mycosphaerella graminicola

  20. The Endo-Arabinanase BcAra1 Is a Novel Host-Specific Virulence Factor of the Necrotic Fungal Phytopathogen Botrytis cinerea

    DEFF Research Database (Denmark)

    Nafisi, Majse; Stranne, Maria; Zhang, Lisha

    2014-01-01

    The plant cell wall is one of the first physical interfaces encountered by plant pathogens and consists of polysaccharides, of which arabinan is an important constituent. During infection, the necrotrophic plant pathogen Botrytis cinerea secretes a cocktail of plant cell-wall-degrading enzymes...

  1. Mycoparasitism of endophytic fungi isolated from reed on soilborne phytopathogenic fungi and production of cell wall-degrading enzymes in vitro.

    Science.gov (United States)

    Cao, Ronghua; Liu, Xiaoguang; Gao, Kexiang; Mendgen, Kurt; Kang, Zhensheng; Gao, Jianfeng; Dai, Yang; Wang, Xue

    2009-12-01

    Antagonism of three endophytic fungi isolated from common reed (Phragmites australis) against eight soilborne pathogenic fungi was investigated on potato dextrose agar by light microscopy, scanning electron microscopy, and transmission electron microscopy. Inhibitory zones were not observed. The microscopical studies suggested that the endophytes inhibit growth of soilborne pathogens by means of coiling around hyphae and, after penetration, the degradation of hyphal cytoplasm. Since penetration of hyphae seems to play a major role in parasitism, we studied the production of cell wall degrading enzymes by the three endophytes. Choiromyces aboriginum produced higher activities of beta-1,3-glucanases compared to Stachybotrys elegans and Cylindrocarpon sp. For C. aboriginum and S. elegans, colloidal chitin was the best substrate for the induction of beta-1,3-glucanases and chitinases, respectively. This result suggests that mycoparasitism by endophytes on soilborne plant pathogens can be explained by their mycoparasitic activity.

  2. Crystallization, data collection and data processing of maltose-binding protein (MalE) from the phytopathogen Xanthomonas axonopodis pv. citri

    International Nuclear Information System (INIS)

    Souza, C. S.; Ferreira, L. C. S.; Thomas, L.; Barbosa, J. A. R. G.; Balan, A.

    2009-01-01

    The Xanthomonas axonopodis pv. citri maltose-binding protein MalE has been crystallized at 293 K using the hanging-drop vapour-diffusion method. Maltose-binding protein is the periplasmic component of the ABC transporter responsible for the uptake of maltose/maltodextrins. The Xanthomonas axonopodis pv. citri maltose-binding protein MalE has been crystallized at 293 K using the hanging-drop vapour-diffusion method. The crystal belonged to the primitive hexagonal space group P6 1 22, with unit-cell parameters a = 123.59, b = 123.59, c = 304.20 Å, and contained two molecules in the asymetric unit. It diffracted to 2.24 Å resolution

  3. Metabolomic-Based Study of the Leafy Gall, the Ecological Niche of the Phytopathogen Rhodococcus fascians, as a Potential Source of Bioactive Compounds

    Directory of Open Access Journals (Sweden)

    Pierre Duez

    2013-06-01

    Full Text Available Leafy gall is a plant hyperplasia induced upon Rhodococcus fascians infection. Previously, by genomic and transcriptomic analysis, it has been reported that, at the early stage of symptom development, both primary and secondary metabolisms are modified. The present study is based on the hypothesis that fully developed leafy gall, could represent a potential source of new bioactive compounds. Therefore, non-targeted metabolomic analysis of aqueous and chloroform extracts of leafy gall and non-infected tobacco was carried out by 1H-NMR coupled to principal component analysis (PCA and orthogonal projections to latent structures-discriminant analysis (OPLS-DA. Polar metabolite profiling reflects modifications mainly in the primary metabolites and in some polyphenolics. In contrast, main modifications occurring in non-polar metabolites concern secondary metabolites, and gas chromatography and mass spectrometry (GC-MS evidenced alterations in diterpenoids family. Analysis of crude extracts of leafy galls and non-infected tobacco leaves exhibited a distinct antiproliferative activity against all four tested human cancer cell lines. A bio-guided fractionation of chloroformic crude extract yield to semi-purified fractions, which inhibited proliferation of glioblastoma U373 cells with IC50 between 14.0 and 2.4 µg/mL. Discussion is focused on the consequence of these metabolic changes, with respect to plant defense mechanisms following infection. Considering the promising role of diterpenoid family as bioactive compounds, leafy gall may rather be a propitious source for drug discovery.

  4. Molecular evolution and phylogenetic analysis of biocontrol genes acquired from SCoT polymorphism of mycoparasitic Trichoderma koningii inhibiting phytopathogen Rhizoctonia solani Kuhn.

    Science.gov (United States)

    Gajera, H P; Hirpara, Darshna G; Katakpara, Zinkal A; Patel, S V; Golakiya, B A

    2016-11-01

    The biocontrol agent Trichoderma (T. harzianum, T. viride, T. virens, T. hamantum, T. koningii, T. pseudokoningii and Trichoderma species) inhibited variably (15.32 - 88.12%) the in vitro growth of Rhizoctonia solani causing root rot in cotton. The T. koningii MTCC 796 evidenced highest (88.12%) growth inhibition of test pathogen followed by T. viride NBAII Tv23 (85.34%). Scanning electron microscopic study confirmed mycoparasitism for MTCC 796 and Tv23 which were capable of completely overgrowing on R. solani by degrading mycelia, coiling around the hyphae with hook-like structures. The antagonists T. harzianum NBAII Th1 and, T. virens NBAII Tvs12 exhibited strong antibiosis and formed 2-4 mm zone of inhibition for 70.28% and 46.62%, respectively growth inhibition of test pathogen. Mycoparasitism is a strong mode of action for biocontrol activity compared with antibiosis. The antagonists Trichoderma strains were performed for start codon targeted (SCoT) polymorphism to acquire biocontrol genes from potent antagonist. The six unique SCoT fragments amplified by genomic DNA of best mycoparasitic antagonist MTCC 796 strain are subjected to DNA sequencing resulted to confirm two functional sequences for activity related to biocontrol genes. The phylogenetic and molecular evolution of functional 824 bp of SCoT-3 (920) and 776 bp of SCoT-6 (806) fragments signify sequence homology with biocontrol genes endochitinase (partial cds of 203 amino acids) and novel hmgR genes (partial cds of 239 amino acids), respectively and the same were annotated and deposited in NCBI GenBank database. The hmgR gene is liable to be express hmg - CoA reductase which is a key enzyme for regulation of terpene biosynthesis and mycoparasitic strains produced triterpenes during antagonism to inhibit growth of fungal pathogen as evidenced with GC-MS profile. The biocontrol genes are found in best antagonist T. koningii MTCC 796 for mycoparasitic activity to restrain the growth of test pathogen R. solani. Copyright © 2016 Elsevier B.V. All rights reserved.

  5. Draft Genome Sequence of Streptomyces sp. Strain Wb2n-11, a Desert Isolate with Broad-Spectrum Antagonism against Soilborne Phytopathogens

    Energy Technology Data Exchange (ETDEWEB)

    Köberl, Martina; White, Richard A.; Erschen, Sabine; El-Arabi, Tarek F.; Jansson, Janet K.; Berg, Gabriele

    2015-08-06

    Streptomyces sp. strain Wb2n-11, isolated from native desert soil, exhibited broad-spectrum antagonism against plant pathogenic fungi, bacteria and nematodes. The 8.2 Mb draft genome reveals genes putatively responsible for its promising biocontrol activity and genes which enable the soil bacterium to directly interact beneficially with plants.

  6. Ultraviolet-Mediated Activation of Photo toxins from Peganum Harmala L. Seedlings to Control both Human-and Phyto-Pathogenic Microorganisms and Tumor Cells

    International Nuclear Information System (INIS)

    Kord, M.; Khafagi, I.; Dewedar, A.

    2003-01-01

    The medicinal plant Peganum harmala L. (zygophyllaceae) contains a number of Beta-carboline alkaloids, which are photosensitizers to bacteria, yeasts and eukaryotic cells in the presence of sunlight and artificial sources of long-wave UV radiation (365 nm). Ultraviolet irradiation of ten-day old aseptically germinated Peganum harmala inoculated on bacterial and yeast bioassay plates elicits strong phototoxic antimicrobials. Callus as well as crude methanol extracts of in vitro cultures were also investigated for the accumulation of photosensitizers. High performance liquid chromatographic analyses of irradiated and control tissues followed by fluorescent detection at 302 nm revealed the formation of serotonin (5-hydroxytryptamine) in irradiated tissues only. Eluted compounds detected at 330 nm revealed more than ten-fold accumulation of harmine, isoharmine and harmol in irradiated tissues. Moreover, several simple beta-carboline alkaloids were produced through irradiation with UV such as harmalanine and harmalacidine. UV-induced phototoxicity was proven against phyto pathogenic bacteria and human-pathogenic bacteria and yeasts. Photo-induced cytotoxicity was observed from two different toxicity bioassays, which are Artemia saline and potato discs tumor assay. The selective UV-dependent biological activities may imply a pharmacological potential of Peganum harmala in the control of infectious diseases and tumor tissues

  7. bcpmr1 encodes a P-type Ca(2+)/Mn(2+)-ATPase mediating cell-wall integrity and virulence in the phytopathogen Botrytis cinerea.

    Science.gov (United States)

    Plaza, Verónica; Lagües, Yanssuy; Carvajal, Mauro; Pérez-García, Luis A; Mora-Montes, Hector M; Canessa, Paulo; Larrondo, Luis F; Castillo, Luis

    2015-03-01

    The cell wall of fungi is generally composed of an inner skeletal layer consisting of various polysaccharides surrounded by a layer of glycoproteins. These usually contain both N- and O-linked oligosaccharides, coupled to the proteins by stepwise addition of mannose residues by mannosyltransferases in the endoplasmic reticulum and the Golgi apparatus. In yeast, an essential luminal cofactor for these mannosyltransferases is Mn(2+) provided by the Ca(2+)/Mn(2+)-ATPase known as Pmr1. In this study, we have identified and characterized the Botrytis cinerea pmr1 gene, the closest homolog of yeast PMR1. We hypothesized that bcpmr1 also encodes a Ca(2+)/Mn(2+)-ATPase that plays an important role in the protein glycosylation pathway. Phenotypic analysis showed that bcpmr1 null mutants displayed a significant reduction in conidial production, radial growth and diameter of sclerotia. Significant alterations in hyphal cell wall composition were observed including a 83% decrease of mannan levels and an increase in the amount of chitin and glucan. These changes were accompanied by a hypersensitivity to cell wall-perturbing agents such as Calcofluor white, Congo red and zymolyase. Importantly, the Δbcpmr1 mutant showed reduced virulence in tomato (leafs and fruits) and apple (fruits) and reduced biofilm formation. Together, our results highlight the importance of bcpmr1 for protein glycosylation, cell wall structure and virulence of B. cinerea. Copyright © 2015 Elsevier Inc. All rights reserved.

  8. Effectiveness of a full-scale horizontal slow sand filter for controlling phytopathogens in recirculating hydroponics: From microbial isolation to full microbiome assessment.

    Science.gov (United States)

    Prenafeta-Boldú, Francesc X; Trillas, Isabel; Viñas, Marc; Guivernau, Miriam; Cáceres, Rafaela; Marfà, Oriol

    2017-12-01

    The microbial disinfestation efficiency of an innovative horizontal-flow slow sand filter (HSSF) for treating nutrient solution spent from an experimental closed-loop nursery was evaluated by means of a combination of culture-dependent and independent molecular techniques. A dense inoculum of the fungal plant pathogen Fusarium oxysporum f.sp. lycopersici was applied in the fertigation system (10 6 cells per mL). Indigenous and introduced populations of eubacteria and fungi were assessed in the nutrient solution, the HSSF influent/effluent, and a sand bed transect by isolation on selective media, as well as by quantitative qPCR and next-generation sequencing (NGS) on target ribosomal genes. The HSSF effectively reduced viable Fusarium propagules and fungal gene content with an efficiency consistently above 99.9% (5 orders of magnitude down). On the other hand, Fusarium cells accumulated in the sand bed, indicating that physical entrapment was the main removal mechanism. The viability of retained Fusarium cells tended to decrease in time, so that treatment efficiency might be enhanced by antagonistic species from the genera Bacillus, Pseudomonas, and Trichoderma, also identified in the sand bed. Indigenous bacterial populations from the HSSF effluent were reduced by 87.2% and 99.9% in terms of colony forming units and gene counts, respectively, when compared to the influent. Furthermore, microbial populations from the HSSF effluent were different from those observed in the sand bed and the influent. In summary, the HSSF microbial disinfestation efficiency is comparable to that reported for other more intensive and costly methodologies, while allowing a significant recovery of water and nutrients. Copyright © 2017 Elsevier B.V. All rights reserved.

  9. Ectopic Expression in Arabidopsis thaliana of an NB-ARC Encoding Putative Disease Resistance Gene from Wild Chinese Vitis pseudoreticulata Enhances Resistance to Phytopathogenic Fungi and Bacteria

    Directory of Open Access Journals (Sweden)

    Zhifeng eWen

    2015-12-01

    Full Text Available Plant resistance proteins mediate pathogen recognition and activate innate immune responses to restrict pathogen proliferation. One common feature of these proteins is an NB-ARC domain. In this study, we characterized a gene encoding a protein with an NB-ARC domain from wild Chinese grapevine Vitis pseudoreticulata accession Baihe-35-1, which was identified in a transcriptome analysis of the leaves following inoculation with Erysiphe necator (Schw., a causal agent of powdery mildew. Transcript levels of this gene, designated VpCN (GenBank accession number KT265084, increased strongly after challenge of grapevine leaves with E. necator. The deduced amino acid sequence was predicted to contain an NB-ARC domain in the C-terminus and an RxCC-like domain similar to CC domain of Rx protein in the N-terminus. Ectopic expression of VpCN in Arabidopsis thaliana resulted in either a wild-type phenotype or a dwarf phenotype. The phenotypically normal transgenic A. thaliana showed enhance resistance to A. thaliana powdery mildew Golovinomyces cichoracearum, as well as to a virulent bacterial pathogen Pseudomonas syringae pv. tomato DC3000. Moreover, promoter::GUS (β-glucuronidase analysis revealed that powdery mildew infection induced the promoter activity of VpCN in grapevine leaves. Finally, a promoter deletion analysis showed that TC rich repeat elements likely play an important role in the response to E. necator infection. Taken together, our results suggest that VpCN contribute to powdery mildew disease resistant in grapevine.

  10. Ectopic Expression in Arabidopsis thaliana of an NB-ARC Encoding Putative Disease Resistance Gene from Wild Chinese Vitis pseudoreticulata Enhances Resistance to Phytopathogenic Fungi and Bacteria.

    Science.gov (United States)

    Wen, Zhifeng; Yao, Liping; Wan, Ran; Li, Zhi; Liu, Chonghuai; Wang, Xiping

    2015-01-01

    Plant resistance proteins mediate pathogen recognition and activate innate immune responses to restrict pathogen proliferation. One common feature of these proteins is an NB-ARC domain. In this study, we characterized a gene encoding a protein with an NB-ARC domain from wild Chinese grapevine Vitis pseudoreticulata accession "Baihe-35-1," which was identified in a transcriptome analysis of the leaves following inoculation with Erysiphe necator (Schw.), a causal agent of powdery mildew. Transcript levels of this gene, designated VpCN (GenBank accession number KT265084), increased strongly after challenge of grapevine leaves with E. necator. The deduced amino acid sequence was predicted to contain an NB-ARC domain in the C-terminus and an RxCC-like domain similar to CC domain of Rx protein in the N-terminus. Ectopic expression of VpCN in Arabidopsis thaliana resulted in either a wild-type phenotype or a dwarf phenotype. The phenotypically normal transgenic A. thaliana showed enhance resistance to A. thaliana powdery mildew Golovinomyces cichoracearum, as well as to a virulent bacterial pathogen Pseudomonas syringae pv. tomato DC3000. Moreover, promoter::GUS (β-glucuronidase) analysis revealed that powdery mildew infection induced the promoter activity of VpCN in grapevine leaves. Finally, a promoter deletion analysis showed that TC rich repeat elements likely play an important role in the response to E. necator infection. Taken together, our results suggest that VpCN contribute to powdery mildew disease resistant in grapevine.

  11. Genomic comparison of the endophyte Herbaspirillum seropedicae SmR1 and the phytopathogen Herbaspirillum rubrisubalbicans M1 by suppressive subtractive hybridization and partial genome sequencing.

    Science.gov (United States)

    Monteiro, Rose A; Balsanelli, Eduardo; Tuleski, Thalita; Faoro, Helison; Cruz, Leonardo M; Wassem, Roseli; de Baura, Valter A; Tadra-Sfeir, Michelle Z; Weiss, Vinícius; DaRocha, Wanderson D; Muller-Santos, Marcelo; Chubatsu, Leda S; Huergo, Luciano F; Pedrosa, Fábio O; de Souza, Emanuel M

    2012-05-01

    Herbaspirillum rubrisubalbicans M1 causes the mottled stripe disease in sugarcane cv. B-4362. Inoculation of this cultivar with Herbaspirillum seropedicae SmR1 does not produce disease symptoms. A comparison of the genomic sequences of these closely related species may permit a better understanding of contrasting phenotype such as endophytic association and pathogenic life style. To achieve this goal, we constructed suppressive subtractive hybridization (SSH) libraries to identify DNA fragments present in one species and absent in the other. In a parallel approach, partial genomic sequence from H. rubrisubalbicans M1 was directly compared in silico with the H. seropedicae SmR1 genome. The genomic differences between the two organisms revealed by SSH suggested that lipopolysaccharide and adhesins are potential molecular factors involved in the different phenotypic behavior. The cluster wss probably involved in cellulose biosynthesis was found in H. rubrisubalbicans M1. Expression of this gene cluster was increased in H. rubrisubalbicans M1 cells attached to the surface of maize root, and knockout of wssD gene led to decrease in maize root surface attachment and endophytic colonization. The production of cellulose could be responsible for the maize attachment pattern of H. rubrisubalbicans M1 that is capable of outcompeting H. seropedicae SmR1. © 2012 Federation of European Microbiological Societies. Published by Blackwell Publishing Ltd. All rights reserved.

  12. Eco-friendly PEG-based controlled release nano-formulations of Mancozeb: Synthesis and bioefficacy evaluation against phytopathogenic fungi Alternaria solani and Sclerotium rolfsii.

    Science.gov (United States)

    Majumder, Sujan; Shakil, Najam A; Kumar, Jitendra; Banerjee, Tirthankar; Sinha, Parimal; Singh, Braj B; Garg, Parul

    2016-12-01

    Controlled release (CR) nano-formulations of Mancozeb (manganese-zinc double salt of N,N-bisdithiocarbamic acid), a protective fungicide, have been prepared using laboratory-synthesized poly(ethylene glycols) (PEGs)-based functionalized amphiphilic copolymers without using any surfactants or external additives. The release kinetics of the developed Mancozeb CR formulations were studied and compared with that of commercially available 42% suspension concentrate and 75% wettable powder. Maximum amount of Mancozeb was released on 42nd day for PEG-600 and octyl chain, PEG-1000 and octyl chain, and PEG-600 and hexadecyl chain, on 35th day for PEG-1000 and hexadecyl chain, on 28th day for PEG-1500 and octyl chain, PEG-2000 and octyl chain, PEG-1500 and hexadecyl chain, and PEG-2000 and hexadecyl chain in comparison to both commercial formulations (15th day). The diffusion exponent (n value) of Mancozeb in water ranged from 0.42 to 0.62 in tested formulations. The half-release (t 1/2 ) values ranged from 17.35 to 35.14 days, and the period of optimum availability of Mancozeb ranged from 18.54 to 35.42 days. Further, the in vitro bioefficacy evaluation of developed formulations was done against plant pathogenic fungi Alternaria solani and Sclerotium rolfsii by poison food technique. Effective dose for 50% inhibition in mgL -1 (ED 50 ) values of developed formulations varied from 1.31 to 2.79 mg L -1 for A. solani, and 1.60 to 3.14 mg L -1 for S. rolfsii. The present methodology is simple, economical, and eco-friendly for the development of environment-friendly CR formulations of Mancozeb. These formulations can be used to optimize the release of Mancozeb to achieve disease control for the desired period depending upon the matrix of the polymer used. Importantly, the maximum amount of active ingredient remains available for a reasonable period after application. In addition, the developed CR formulations were found to be suitable for fungicidal applications, allowing use of Mancozeb in lower doses.

  13. A eukaryotic-acquired gene by a biotrophic phytopathogen allows prolonged survival on the host by counteracting the shut-down of plant photosynthesis

    KAUST Repository

    Garavaglia, Betiana S.

    2010-01-28

    Xanthomonas citri pv. citri, the bacteria responsible for citrus canker posses a biological active plant natriuretic peptide (PNP)-like protein, not present in any other bacteria. PNPs are a class of extracellular, systemically mobile peptides that elicit a number of plant responses important in homeostasis and growth. Previously, we showed that a Xanthomonas citri pv. citri mutant lacking the PNP-like protein XacPNP produced more necrotic lesions in citrus leaves than wild type infections and suggested a role for XacPNP in the regulation of host homeostasis. Here we have analyzed the proteome modifications observed in citrus leaves infected with the wild type and XacPNP deletion mutant bacteria. While both of them cause downregulation of enzymes related to photosynthesis as well as chloroplastic ribosomal proteins, proteins related to defense responses are up-regulated. However, leaves infiltrated with the XacPNP deletion mutant show a more pronounced decrease in photosynthetic proteins while no reduction in defense related proteins as compared to the wild-type pathogen. This suggests that XacPNP serves the pathogen to maintain host photosynthetic efficiency during pathogenesis. The results from the proteomics analyses are consistent with our chlorophyll fluorescence data and transcript analyses of defense genes that show a more marked reduction in photosynthesis in the mutant but no difference in the induction of genes diagnostic for biotic-stress responses. We therefore conclude that XacPNP counteracts the shut-down of host photosynthesis during infection and in that way maintains the tissue in better conditions, suggesting that the pathogen has adapted a host gene to modify its natural host and render it a better reservoir for prolonged bacterial survival and thus for further colonization. 2010 Garavaglia et al.

  14. Genomic overview of the phytopathogen Pectobacterium wasabiae strain RNS 08.42.1A suggests horizontal acquisition of quorum-sensing genes.

    Science.gov (United States)

    Khayi, Slimane; Raoul des Essarts, Yannick; Quêtu-Laurent, Angélique; Moumni, Mohieddine; Hélias, Valérie; Faure, Denis

    2015-04-01

    The blackleg and soft-rot diseases caused by pectinolytic enterobacteria such as Pectobacterium and Dickeya are major causes of losses affecting potato crop in the field and upon storage. In this work, we report the isolation, characterization and genome analysis of the Pectobacterium wasabiae (formerly identified as Pectobacterium carotovorum subsp. carotovorum) strain RNS 08.42.1A, that has been isolated from a Solanum tuberosum host plant in France. Comparative genomics with 3 other P. wasabiae strains isolated from potato plants in different areas in North America and Europe, highlighted both a strong similarity at the whole genome level (ANI > 99 %) and a conserved synteny of the virulence genes. In addition, our analyses evidenced a robust separation between these four P. wasabiae strains and the type strain P. wasabiae CFBP 3304(T), isolated from horseradish in Japan. In P. wasabiae RNS 08.42.1A, the expI and expR nucleotidic sequences are more related to those of some Pectobacterium atrosepticum and P. carotovorum strains (90 % of identity) than to those of the other potato P. wasabiae strains (70 to 74 % of identity). This could suggest a recruitment of these genes in the P. wasabiae strain RNS 08.42.1A by an horizontal transfer between pathogens infecting the same potato host plant.

  15. A novel nonsegmented double-stranded RNA mycovirus identified in the phytopathogenic fungus Nigrospora oryzae shows similarity to partitivirus-like viruses.

    Science.gov (United States)

    Zhou, Qian; Zhong, Jie; Hu, Yue; Da Gao, Bi

    2016-01-01

    Nigrospora oryzae is a pathogen that can infect plants of various species. Here, we report the isolation of a novel mycovirus from N. oryzae infecting rice, as well as the complete genome sequence and genomic organization of this virus, which we have named "Nigrospora oryzae nonsegmented RNA virus 1" (NoNRV1). The genome of NoNRV1 contained two non-overlapping open reading frames (ORF1 and ORF2) potentially encoding a protein with an unknown function in ORF1 and a putative RNA-dependent RNA polymerase (RdRp) in ORF2. Homology and phylogenetic analysis revealed that NoNRV1 was most similar to the Ustilaginoidea virens nonsegmented virus 1 (UvNV-1) and distantly related to members of the virus family Partitiviridae. It is proposed that NoNRV1, together with UvNV-1 and other related viruses, might represent a novel virus taxon of mycoviruses belonging to a partitivirus-like lineage.

  16. COMPARATIVE GENOME ANALYSES OF MONILIOPHTHORA PERNICIOSA AND MONILIOPHTHORA RORERI: TWO CLOSELY RELATED PHYTOPATHOGENIC BASIDIOMYCETES THAT CAUSE DISTINCTLY DIFFERENT DISEASES OF THEOBROMA CACAO

    Science.gov (United States)

    Theobroma cacao (cacao), the source of chocolate, is a tropical understory tree. Fungal diseases such as Witches’ Broom Disease (WBD) and Frosty Pod Rot Disease (FPRD) of cacao have devastated cacao production in much of the Western Hemisphere and are threats to the main cacao producing regions in A...

  17. Application of Silver Nanostructures Synthesized by Cold Atmospheric Pressure Plasma for Inactivation of Bacterial Phytopathogens from the Genera Dickeya and Pectobacterium

    Directory of Open Access Journals (Sweden)

    Anna Dzimitrowicz

    2018-02-01

    Full Text Available Pectinolytic bacteria are responsible for significant economic losses by causing diseases on numerous plants. New methods are required to control and limit their spread. One possibility is the application of silver nanoparticles (AgNPs that exhibit well-established antibacterial properties. Here, we synthesized AgNPs, stabilized by pectins (PEC or sodium dodecyl sulphate (SDS, using a direct current atmospheric pressure glow discharge (dc-APGD generated in an open-to-air and continuous-flow reaction-discharge system. Characterization of the PEC-AgNPs and SDS-AgNPs with UV/Vis absorption spectroscopy, transmission electron microscopy, energy dispersive X-ray spectroscopy, and selected area electron diffraction revealed the production of spherical, well dispersed, and face cubic centered crystalline AgNPs, with average sizes of 9.33 ± 3.37 nm and 28.3 ± 11.7 nm, respectively. Attenuated total reflection-Fourier transformation infrared spectroscopy supported the functionalization of the nanostructures by PEC and SDS. Antibacterial activity of the AgNPs was tested against Dickeya spp. and Pectobacterium spp. strains. Both PEC-AgNPs and SDS-AgNPs displayed bactericidal activity against all of the tested isolates, with minimum inhibitory concentrations of 5.5 mg∙L−1 and 0.75–3 mg∙L−1, respectively. The collected results suggest that the dc-APGD reaction-discharge system can be applied for the production of defined AgNPs with strong antibacterial properties, which may be further applied in plant disease management.

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

    Science.gov (United States)

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

    2013-06-01

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

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

    Science.gov (United States)

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

  20. Geosmithia morbida sp nov., a new phytopathogenic species living in symbiosis with the walnut twig beetle (Pityophthorus juglandis) on Juglans in USA

    Czech Academy of Sciences Publication Activity Database

    Kolařík, Miroslav; Freeland, E.

    2011-01-01

    Roč. 103, č. 2 (2011), s. 325-332 ISSN 0027-5514 R&D Projects: GA ČR GP206/08/P322 Institutional research plan: CEZ:AV0Z50200510 Keywords : Geosmithia * insect-associated fungi * thousand cankers disease Subject RIV: EE - Microbiology, Virology Impact factor: 2.031, year: 2011

  1. Virulence, Host-Selective Toxin Production, and Development of Three Cochliobolus Phytopathogens Lacking the Sfp-Type 4'-Phosphopantetheinyl Transferase Ppt1.

    Science.gov (United States)

    Zainudin, Nur Ain Izzati Mohd; Condon, Bradford; De Bruyne, Lieselotte; Van Poucke, Christof; Bi, Qing; Li, Wei; Höfte, Monica; Turgeon, B Gillian

    2015-10-01

    The Sfp-type 4'-phosphopantetheinyl transferase Ppt1 is required for activation of nonribosomal peptide synthetases, including α-aminoadipate reductase (AAR) for lysine biosynthesis and polyketide synthases, enzymes that biosynthesize peptide and polyketide secondary metabolites, respectively. Deletion of the PPT1 gene, from the maize pathogen Cochliobolus heterostrophus and the rice pathogen Cochliobolus miyabeanus, yielded strains that were significantly reduced in virulence to their hosts. In addition, ppt1 mutants of C. heterostrophus race T and Cochliobolus victoriae were unable to biosynthesize the host-selective toxins (HST) T-toxin and victorin, respectively, as judged by bioassays. Interestingly, ppt1 mutants of C. miyabeanus were shown to produce tenfold higher levels of the sesterterpene-type non-HST ophiobolin A, as compared with the wild-type strain. The ppt1 strains of all species were also reduced in tolerance to oxidative stress and iron depletion; both phenotypes are associated with inability to produce extracellular siderophores biosynthesized by the nonribosomal peptide synthetase Nps6. Colony surfaces were hydrophilic, a trait previously associated with absence of C. heterostrophus Nps4. Mutants were decreased in asexual sporulation and C. heterostrophus strains were female-sterile in sexual crosses; the latter phenotype was observed previously with mutants lacking Nps2, which produces an intracellular siderophore. As expected, mutants were albino, since they cannot produce the polyketide melanin and were auxotrophic for lysine because they lack an AAR.

  2. Are Epiphytic Microbial Communities in the Carposphere of Ripening Grape Clusters (Vitis vinifera L.) Different between Conventional, Organic, and Biodynamic Grapes?

    Science.gov (United States)

    Kecskeméti, Elizabeth; Berkelmann-Löhnertz, Beate; Reineke, Annette

    2016-01-01

    Using barcoded pyrosequencing fungal and bacterial communities associated with grape berry clusters (Vitis vinifera L.) obtained from conventional, organic and biodynamic vineyard plots were investigated in two subsequent years at different stages during berry ripening. The four most abundant operational taxonomic units (OTUs) based on fungal ITS data were Botrytis cinerea, Cladosporium spp., Aureobasidium pullulans and Alternaria alternata which represented 57% and 47% of the total reads in 2010 and 2011, respectively. Members of the genera Sphingomonas, Gluconobacter, Pseudomonas, Erwinia, and Massilia constituted 67% of the total number of bacterial 16S DNA reads in 2010 samples and 78% in 2011 samples. Viticultural management system had no significant effect on abundance of fungi or bacteria in both years and at all three sampling dates. Exceptions were A. alternata and Pseudomonas spp. which were more abundant in the carposphere of conventional compared to biodynamic berries, as well as Sphingomonas spp. which was significantly less abundant on conventional compared to organic berries at an early ripening stage in 2011. In general, there were no significant differences in fungal and bacterial diversity indices or richness evident between management systems. No distinct fungal or bacterial communities were associated with the different maturation stages or management systems, respectively. An exception was the last stage of berry maturation in 2011, where the Simpson diversity index was significantly higher for fungal communities on biodynamic compared to conventional grapes. Our study highlights the existence of complex and dynamic microbial communities in the grape cluster carposphere including both phytopathogenic and potentially antagonistic microorganisms that can have a significant impact on grape production. Such knowledge is particularly relevant for development, selection and application of effective control measures against economically important

  3. 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.

  4. The bile acid deoxycholate elicits defences in Arabidopsis and reduces bacterial infection.

    Science.gov (United States)

    Zarattini, Marco; Launay, Alban; Farjad, Mahsa; Wénès, Estelle; Taconnat, Ludivine; Boutet, Stéphanie; Bernacchia, Giovanni; Fagard, Mathilde

    2017-05-01

    Disease has an effect on crop yields, causing significant losses. As the worldwide demand for agricultural products increases, there is a need to pursue the development of new methods to protect crops from disease. One mechanism of plant protection is through the activation of the plant immune system. By exogenous application, 'plant activator molecules' with elicitor properties can be used to activate the plant immune system. These defence-inducing molecules represent a powerful and often environmentally friendly tool to fight pathogens. We show that the secondary bile acid deoxycholic acid (DCA) induces defence in Arabidopsis and reduces the proliferation of two bacterial phytopathogens: Erwinia amylovora and Pseudomonas syringae pv. tomato. We describe the global defence response triggered by this new plant activator in Arabidopsis at the transcriptional level. Several induced genes were selected for further analysis by quantitative reverse transcription-polymerase chain reaction. We describe the kinetics of their induction and show that abiotic stress, such as moderate drought or nitrogen limitation, does not impede DCA induction of defence. Finally, we investigate the role in the activation of defence by this bile acid of the salicylic acid biosynthesis gene SID2, of the receptor-like kinase family genes WAK1-3 and of the NADPH oxidase-encoding RbohD gene. Altogether, we show that DCA constitutes a promising molecule for plant protection which can induce complementary lines of defence, such as callose deposition, reactive oxygen species accumulation and the jasmonic acid and salicylic acid signalling pathways. © 2016 BSPP AND JOHN WILEY & SONS LTD.

  5. Are Epiphytic Microbial Communities in the Carposphere of Ripening Grape Clusters (Vitis vinifera L.) Different between Conventional, Organic, and Biodynamic Grapes?

    Science.gov (United States)

    Kecskeméti, Elizabeth; Berkelmann-Löhnertz, Beate; Reineke, Annette

    2016-01-01

    Using barcoded pyrosequencing fungal and bacterial communities associated with grape berry clusters (Vitis vinifera L.) obtained from conventional, organic and biodynamic vineyard plots were investigated in two subsequent years at different stages during berry ripening. The four most abundant operational taxonomic units (OTUs) based on fungal ITS data were Botrytis cinerea, Cladosporium spp., Aureobasidium pullulans and Alternaria alternata which represented 57% and 47% of the total reads in 2010 and 2011, respectively. Members of the genera Sphingomonas, Gluconobacter, Pseudomonas, Erwinia, and Massilia constituted 67% of the total number of bacterial 16S DNA reads in 2010 samples and 78% in 2011 samples. Viticultural management system had no significant effect on abundance of fungi or bacteria in both years and at all three sampling dates. Exceptions were A. alternata and Pseudomonas spp. which were more abundant in the carposphere of conventional compared to biodynamic berries, as well as Sphingomonas spp. which was significantly less abundant on conventional compared to organic berries at an early ripening stage in 2011. In general, there were no significant differences in fungal and bacterial diversity indices or richness evident between management systems. No distinct fungal or bacterial communities were associated with the different maturation stages or management systems, respectively. An exception was the last stage of berry maturation in 2011, where the Simpson diversity index was significantly higher for fungal communities on biodynamic compared to conventional grapes. Our study highlights the existence of complex and dynamic microbial communities in the grape cluster carposphere including both phytopathogenic and potentially antagonistic microorganisms that can have a significant impact on grape production. Such knowledge is particularly relevant for development, selection and application of effective control measures against economically important

  6. The public health risks of enterobacterial isolates producing extended-spectrum Beta-lactamases (ESBL) or AmpC Beta-lactamases in food and food-producing animals: An EU perspective of epidemiology, analytical methods, risk factors and control options

    NARCIS (Netherlands)

    Liebana, E.; Carattoli, A.; Coque, T.M.; Hasman, H.; Magiorakos, A.P.; Mevius, D.J.; Peixe, L.; Schuepbach-Regula, G.; Torneke, K.; Torren-Edo, J.; Torres, C.; Threlfall, J.

    2013-01-01

    blaESBL and blaAmpC in Enterobacteriaceae are spread by plasmid-mediated integrons, insertion sequences and transposons, some of which are homologous in bacteria from food-animals, foods and humans. These genes have been frequently identified in Escherichia coli and Salmonella from food-animals; the

  7. ORF Alignment: NC_004547 [GENIUS II[Archive

    Lifescience Database Archive (English)

    Full Text Available erase ... [Erwinia carotovora subsp. atroseptica SCRI1043] ... Length = 142 ... Query: 10 ... RPEYKDRAIHYFQRHWASDETLILYEDCISH...CIDAENPLPDWYLMEKEGDIIGGAGLITN 69 ... RPEYKDRAIHYFQRHWASDETLILYEDCISH...CIDAENPLPDWYLMEKEGDIIGGAGLITN Sbjct: 1 ... RPEYKDRAIHYFQRHWASDETLILYEDCISHCIDAENPLPDWYLMEKE

  8. The Immunogenicity of a Biological Simulant: Strategies for the Improvement of Antibody-Based Detection

    National Research Council Canada - National Science Library

    Grahame, David A; Gencic, Simonida; Bronk, Burt V

    2005-01-01

    .... The bacterium Pantoea agglomerans (formerly Erwinia herbicola, Eh) presently is used to simulate vegetative biological agents, however, anti-Eh antibodies of high affinity and specificity are needed...

  9. Pantoea applied genomics to understand and improve biocontrol activity against fire blight

    Science.gov (United States)

    Pantoea agglomerans and P. vagans (ex. Erwinia herbicola) are common epiphytes of pome fruit flowers and three strains (E325, P10c, C9-1) have been commercially developed as effective biocontrol products for managing fire blight (Erwinia amylovora). Antibiotics as a standard, reliable chemical optio...

  10. Development of species-specific DNA probes for Campylobacter jejuni, Campylobacter coli, and Campylobacter lari by polymerase chain reaction fingerprinting

    NARCIS (Netherlands)

    Giesendorf, B A; van Belkum, A; Koeken, A; Stegeman, H; Henkens, M H; van der Plas, J; Goossens, H; Niesters, H G; Quint, W G

    The application of polymerase chain reaction (PCR) fingerprinting assays enables discrimination between species and strains of microorganisms. PCR primers aiming at arbitrary sequences in combination with primers directed against the repetitive extragenic palindrome (REP) or enterobacterial

  11. Research

    African Journals Online (AJOL)

    abp

    2015-06-09

    ;Enterobacter 22.2 for IP and 50% for OP. Conclusion: High resistance rates to quinolones were observed not only for in-patients but also for out-patients with urinary tract enterobacterial infections. These findings demonstrate ...

  12. Enteroantigen-presenting B cells efficiently stimulate CD4(+) T cells in vitro

    DEFF Research Database (Denmark)

    Schmidt, Esben Gjerløff Wedebye; Kristensen, Nanna Ny; Claesson, Mogens Helweg

    2011-01-01

    Presentation of enterobacterial antigens by antigen-presenting cells and activation of enteroantigen-specific CD4(+) T cells are considered crucial steps in inflammatory bowel disease (IBD) pathology. The detrimental effects of such CD4(+) T cells have been thoroughly demonstrated in models...... of colitis. Also, we have previously established an in vitro assay where murine enteroantigen-specific colitogenic CD4(+) CD25(-) T cells are activated by splenocytes pulsed with an enterobacterial extract....

  13. Development of a Rhizoctonia solani AG1-IB Specific Gene Model Enables Comparative Genome Analyses between Phytopathogenic R. solani AG1-IA, AG1-IB, AG3 and AG8 Isolates.

    Directory of Open Access Journals (Sweden)

    Daniel Wibberg

    Full Text Available Rhizoctonia solani, a soil-born plant pathogenic basidiomycetous fungus, affects various economically important agricultural and horticultural crops. The draft genome sequence for the R. solani AG1-IB isolate 7/3/14 as well as a corresponding transcriptome dataset (Expressed Sequence Tags--ESTs were established previously. Development of a specific R. solani AG1-IB gene model based on GMAP transcript mapping within the eukaryotic gene prediction platform AUGUSTUS allowed detection of new genes and provided insights into the gene structure of this fungus. In total, 12,616 genes were recognized in the genome of the AG1-IB isolate. Analysis of predicted genes by means of different bioinformatics tools revealed new genes whose products potentially are involved in degradation of plant cell wall components, melanin formation and synthesis of secondary metabolites. Comparative genome analyses between members of different R. solani anastomosis groups, namely AG1-IA, AG3 and AG8 and the newly annotated R. solani AG1-IB genome were performed within the comparative genomics platform EDGAR. It appeared that only 21 to 28% of all genes encoded in the draft genomes of the different strains were identified as core genes. Based on Average Nucleotide Identity (ANI and Average Amino-acid Identity (AAI analyses, considerable sequence differences between isolates representing different anastomosis groups were identified. However, R. solani isolates form a distinct cluster in relation to other fungi of the phylum Basidiomycota. The isolate representing AG1-IB encodes significant more genes featuring predictable functions in secondary metabolite production compared to other completely sequenced R. solani strains. The newly established R. solani AG1-IB 7/3/14 gene layout now provides a reliable basis for post-genomics studies.

  14. Production of extracellular lipase by the phytopathogenic fungus Fusarium solani FS1 Produção de lipase extracelular pelo fungo fitopatogênico Fusarium solani FS1

    Directory of Open Access Journals (Sweden)

    Maria de Mascena Diniz Maia

    1999-12-01

    Full Text Available A Brazilian strain of Fusarium solani was tested for extracellular lipase production in peptone-olive oil medium. The fungus produced 10,500 U.l-1 of lipase after 72 hours of cultivation at 25oC in shake-flask at 120 rpm in a medium containing 3% (w/v peptone plus 0.5% (v/v olive oil. Glucose (1% w/v was found to inhibit the inductive effect of olive oil. Peptone concentrations below 3% (w/v resulted in a reduced lipase production while increased olive oil concentration (above 0.5% did not further stimulate lipase production. The optimum lipase activity was achieved at pH 8.6 and 30oC and a good enzyme stability (80% activity retention was observed at pH ranging from 7.6 to 8.6, and the activity rapidly dropped at temperatures above 50oC. Lipase activity was stimulated by the addition of n-hexane to the culture medium supernatants, in contrast to incubation with water-soluble solvents.

  15. Production of extracellular lipase by the phytopathogenic fungus Fusarium solani FS1 Produção de lipase extracelular pelo fungo fitopatogênico Fusarium solani FS1

    OpenAIRE

    Maria de Mascena Diniz Maia; Marcia Maria Camargo de Morais; Marcos Antonio de Morais Jr.; Eduardo Henrique Magalhães Melo; José Luiz de Lima Filho

    1999-01-01

    A Brazilian strain of Fusarium solani was tested for extracellular lipase production in peptone-olive oil medium. The fungus produced 10,500 U.l-1 of lipase after 72 hours of cultivation at 25oC in shake-flask at 120 rpm in a medium containing 3% (w/v) peptone plus 0.5% (v/v) olive oil. Glucose (1% w/v) was found to inhibit the inductive effect of olive oil. Peptone concentrations below 3% (w/v) resulted in a reduced lipase production while increased olive oil concentration (above 0.5%) did n...

  16. Constitutive heterologous overexpression of a TIR-NB-ARC-LRR gene encoding a putative disease resistance protein from wild Chinese Vitis pseudoreticulata in Arabidopsis and tobacco enhances resistance to phytopathogenic fungi and bacteria.

    Science.gov (United States)

    Wen, Zhifeng; Yao, Liping; Singer, Stacy D; Muhammad, Hanif; Li, Zhi; Wang, Xiping

    2017-03-01

    Plants use resistance (R) proteins to detect pathogen effector proteins and activate their innate immune response against the pathogen. The majority of these proteins contain an NB-ARC (nucleotide-binding adaptor shared by APAF-1, R proteins, and CED-4) domain along with a leucine-rich repeat (LRR), and some also bear a toll interleukin 1 receptor (TIR) domain. In this study, we characterized a gene encoding a TIR-NB-ARC-LRR R protein (VpTNL1) (GenBank accession number KX649890) from wild Chinese grapevine Vitis pseudoreticulata accession "Baihe-35-1", which was identified previously from a transcriptomic analysis of leaves inoculated with powdery mildew (PM; Erysiphe necator (Schw.)). The VpTNL1 transcript was found to be highly induced in V. pseudoreticulata following inoculation with E. necator, as well as treatment with salicylic acid (SA). Sequence analysis demonstrated that the deduced amino acid sequence contained a TIR domain at the N-terminus, along with an NB-ARC and four LRRs domains within the C-terminus. Constitutive expression of VpTNL1 in Arabidopsis thaliana resulted in either a wild-type or dwarf phenotype. Intriguingly, the phenotypically normal transgenic lines displayed enhanced resistance to Arabidopsis PM, Golovinomyces cichoracearum, as well as to the virulent bacterial pathogen Pseudomonas syringae pv. tomato DC3000. Similarly, constitutive expression of VpTNL1 in Nicotiana tabacum was found to confer enhanced resistance to tobacco PM, Erysiphe cichoacearum DC. Subsequent isolation of the VpTNL1 promoter and deletion analysis indicated that TC-rich repeats and TCA elements likely play an important role in its response to E. necator and SA treatment, respectively. Taken together, these results indicate that VpTNL1 contributes to PM resistance in grapevine and provide an interesting gene target for the future amelioration of grape via breeding and/or biotechnology. Copyright © 2017 Elsevier Masson SAS. All rights reserved.

  17. Evaluación biológica del manejo de picudos y nematodos fitopatógenos en plátano (Musa AAB Biological evaluation of the management of borers and phytopathogenic nematodes of plantain (Musa AAB

    Directory of Open Access Journals (Sweden)

    Carolina González Cardona

    2009-10-01

    Full Text Available El trabajo se desarrolló en la granja Montelindo, municipio de Palestina (Caldas a 5° 05' N y 75° 40' O, a 1010 m.s.n.m., 23.5 °C, precipitación anual de 2100 mm y humedad relativa de 76%, con el fin de generar información sobre el manejo de picudos y nematodos fitoparásitos del plátano. Se usó un diseño en bloques completos al azar con cuatro tratamientos por bloque, tres repeticiones y 24 plantas por repetición. Para el manejo de los picudos se hicieron aplicaciones de Carbofurán, Beauveria bassiana y Metarhizium anisopliae en trampas tipo columna. Para el control de nematodos se hicieron aplicaciones en el suelo de Carbofurán y dos cepas comerciales de Paecilomyces lilacinus. Se evaluaron el número de adultos de picudos en trampas, la infección de estos por los hongos empleados y la población de nematodos en suelo y raíces. Se encontró que las trampas tratadas con Carbofurán fueron significativamente más efectivas para la captura de insectos. En laboratorio se estableció que M. anisopliae tuvo una mejor capacidad para infectar adultos del insecto en el campo. La población de nematodos fue menor en suelo y raíces de las plantas tratadas con Carbofurán. Paecilomyces lilacinus no fue efectivo para reducir las poblaciones de nematodos. Los géneros de nematodos predominantes fueron Radopholus, Pratylenchus, Meloidogyne y Helicotylenchus.This work was carried out at the ‘Montelindo’ farm, municipality of Palestina (Department of Caldas, Colombia, located at 5° 05' N and 75° 40' W, at 1010 m.a.s.l., 23.5 °C, with 2100 mm of annual rainfall, and relative humidity of 76%, in order to generate information on the management of borers and parasitic nematodes of the plantain. A completely randomised block experimental design was used, with four treatments per block, three replicates and 24 plants per replicate. For the management of borers, applications of Carbofuran, Beauveria bassiana and Metarhizium anisopliae were made into column type traps. To control nematodes soil applications of Carbofuran and two commercial strains of Paecilomyces linacinus were made. The number of adult borers in the traps, the infection of these by the fungi used, and the population of nematodes in both soil and roots were evaluated. It was found that the traps treated with Carbofuran were significantly more effective for insect capture. In the lab, it was established that M. anisopliae had better capacity to infect adult insects in the field. P. lilacinus was ineffective at reducing nematode populations. The nematode population was lower in soil and roots of plants treated with Carbofuran. The predominant genera of nematodes were Radopholus, Pratylenchus, Meloidogyne and Helicotylenchus.

  18. A specific interdomain interaction preserves the structural and binding properties of the ModA protein from the phytopathogen Xanthomonas citri domain interaction and transport in ModA.

    Science.gov (United States)

    Santacruz-Perez, Carolina; Pegos, Vanessa Rodrigues; Honorato, Rodrigo V; Verli, Hugo; Lindahl, Erik; Barbosa, João Alexandre Ribeiro Gonçalves; Balan, Andrea

    2013-11-01

    The periplasmic-binding proteins in ATP-binding cassette systems (ABC Transporters) are responsible for the capture and delivery of ligands to their specific transporters, triggering a series of ATP-driven conformational changes that leads to the transport of the ligand. Structurally consisting of two lobes, the proteins change conformation after interaction with the ligand. The structure of the molybdate-binding protein (ModA) from Xanthomonas citri, bound to molybdate, was previously solved by our group and an interdomain interaction, mediated by a salt bridge between K127 and D59, apparently supports the binding properties and keeps the domains closed. To determinate the importance of this interaction, we built two ModA mutants, K127S and D59A, and analysed their functional and structural properties. Based on a set of spectroscopic experiments, crystallisation trials, structure determination and molecular dynamics (MD) simulations, we showed that the salt bridge is essential to maintain the structure and binding properties. Additionally, the MD simulations revealed that this mutant adopted a more compact structure that packed down the ligand-binding pocket. From the closed bound to open structure, the positioning of the helices forming the dipole and the salt bridge are essential to induce an intermediate state. Copyright © 2013 Elsevier Inc. All rights reserved.

  19. Draft Genomes, Phylogenetic Reconstruction, and Comparative Genomics of Two Novel Cohabiting Bacterial Symbionts Isolated from Frankliniella occidentalis.

    Science.gov (United States)

    Facey, Paul D; Méric, Guillaume; Hitchings, Matthew D; Pachebat, Justin A; Hegarty, Matt J; Chen, Xiaorui; Morgan, Laura V A; Hoeppner, James E; Whitten, Miranda M A; Kirk, William D J; Dyson, Paul J; Sheppard, Sam K; Del Sol, Ricardo

    2015-07-15

    Obligate bacterial symbionts are widespread in many invertebrates, where they are often confined to specialized host cells and are transmitted directly from mother to progeny. Increasing numbers of these bacteria are being characterized but questions remain about their population structure and evolution. Here we take a comparative genomics approach to investigate two prominent bacterial symbionts (BFo1 and BFo2) isolated from geographically separated populations of western flower thrips, Frankliniella occidentalis. Our multifaceted approach to classifying these symbionts includes concatenated multilocus sequence analysis (MLSA) phylogenies, ribosomal multilocus sequence typing (rMLST), construction of whole-genome phylogenies, and in-depth genomic comparisons. We showed that the BFo1 genome clusters more closely to species in the genus Erwinia, and is a putative close relative to Erwinia aphidicola. BFo1 is also likely to have shared a common ancestor with Erwinia pyrifoliae/Erwinia amylovora and the nonpathogenic Erwinia tasmaniensis and genetic traits similar to Erwinia billingiae. The BFo1 genome contained virulence factors found in the genus Erwinia but represented a divergent lineage. In contrast, we showed that BFo2 belongs within the Enterobacteriales but does not group closely with any currently known bacterial species. Concatenated MLSA phylogenies indicate that it may have shared a common ancestor to the Erwinia and Pantoea genera, and based on the clustering of rMLST genes, it was most closely related to Pantoea ananatis but represented a divergent lineage. We reconstructed a core genome of a putative common ancestor of Erwinia and Pantoea and compared this with the genomes of BFo bacteria. BFo2 possessed none of the virulence determinants that were omnipresent in the Erwinia and Pantoea genera. Taken together, these data are consistent with BFo2 representing a highly novel species that maybe related to known Pantoea. © The Author(s) 2015. Published by

  20. Antibacterial activity of Boenninghausenia albiflora Reichb ...

    African Journals Online (AJOL)

    ... maceration were screened for their antimicrobial activity against eight animal and plant pathogenic bacteria (Bacillus subtilis, Staphylococcus aureus, Erwinia chrysanthemi, Escherichia coli, Proteus vulgaris, Xanthomonas phaseoli, Agrobacterium tumefaciens and Xanthomonas campestris) using disc diffusion method.