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Sample records for cinerea xylanase xyn11a

  1. The Botrytis cinerea xylanase Xyn11A contributes to virulence with its necrotizing activity, not with its catalytic activity

    Directory of Open Access Journals (Sweden)

    González Celedonio

    2010-02-01

    Full Text Available Abstract Background The Botrytis cinerea xylanase Xyn11A has been previously shown to be required for full virulence of this organism despite its poor contribution to the secreted xylanase activity and the low xylan content of B. cinerea hosts. Intriguingly, xylanases from other fungi have been shown to have the property, independent of the xylan degrading activity, to induce necrosis when applied to plant tissues, so we decided to test the hypothesis that secreted Xyn11A contributes to virulence by promoting the necrosis of the plant tissue surrounding the infection, therefore facilitating the growth of this necrotroph. Results We show here that Xyn11A has necrotizing activity on plants and that this capacity is conserved in site-directed mutants of the protein lacking the catalytic activity. Besides, Xyn11A contributes to the infection process with the necrotizing and not with the xylan hydrolyzing activity, as the catalytically-impaired Xyn11A variants were able to complement the lower virulence of the xyn11A mutant. The necrotizing activity was mapped to a 30-amino acids peptide in the protein surface, and this region was also shown to mediate binding to tobacco spheroplasts by itself. Conclusions The main contribution of the xylanase Xyn11A to the infection process of B. cinerea is to induce necrosis of the infected plant tissue. A conserved 30-amino acids region on the enzyme surface, away from the xylanase active site, is responsible for this effect and mediates binding to plant cells.

  2. Conservation of XYN11A and XYN11B xylanase genes in Bipolaris sorghicola, Cochliobolus sativus, Cochliobolus heterostrophus, and Cochliobolus spicifer.

    Science.gov (United States)

    Emami, Kaveh; Hack, Ethan

    2002-10-01

    Two types of xylanase gene, XYN11A ( XYL1) and XYN11B ( XYL2), were amplified by PCR and partially sequenced in four phytopathogenic species of the ascomycete fungal genus Cochliobolus (anamorph genus Bipolaris). Three of the species, C. heterostrophus ( B. maydis), C. sativus ( B. sorokiniana), and Bipolaris sorghicola (no teleomorph known), are interrelated; the fourth, C. spicifer ( B. spicifera), was found, through analysis of the 5.8S RNA and internal transcribed spacer (ITS) sequences of its ribosomal DNA, to be more distantly related to the other three. Isolates from all four species contain orthologous XYN11A and XYN11B genes, but a set of laboratory strains of C. heterostrophus gave no product corresponding to the XYN11B gene. The patterns of evolution of the two xylanase genes and ribosomal DNA sequences are mutually consistent; the results indicate that the two genes were present in the common ancestor of all Cochliobolus species and are evolving independently of each other.

  3. Synergistic effect and application of xylanases as accessory enzymes to enhance the hydrolysis of pretreated bagasse.

    Science.gov (United States)

    Gonçalves, Geisa A L; Takasugi, Yusaku; Jia, Lili; Mori, Yutaro; Noda, Shuhei; Tanaka, Tsutomu; Ichinose, Hirofumi; Kamiya, Noriho

    2015-05-01

    Recently, the new trend in the second-generation ethanol industry is to use mild pretreatments, in order to reduce costs and to keep higher content of hemicellulose in the biomass. Nevertheless, a high enzyme dosage is still required in the conversion of (hemi)cellulose. The interaction between cellulases and xylanases seems to be an effective alternative to reduce enzyme loading in the saccharification process. At first, to evaluate the synergism of xylanases on bagasse degradation, we have produced two xylanases from glycoside hydrolase family 10 (GH10) and three xylanases from glycoside hydrolase family 11 (GH11), from two thermophilic organisms, Thermobifida fusca and Clostridium thermocellum, and one mesophilic organism, Streptomyces lividans. Peracetic acid (PAA) pretreated bagasse was used as substrate. The combination of XynZ-C (GH10, from C. thermocellum), and XlnB (GH11, from S. lividans) presented the highest degree of synergy after 6h (3.62). However, the combination of XynZ-C and Xyn11A (GH11, from T. fusca) resulted in the highest total yield of reducing sugars. To evaluate the synergism between xylanases and cellulases, commercial cellulase preparation from Trichoderma reesei was combined with the selected xylanases, XynZ-C and Xyn11A. About 2-fold increase was observed in the concentration of reducing sugars, when both xylanases, XynZ-C and Xyn11A, were added together with T. reesei cellulases in the reaction mixture.

  4. [Occupational allergies to xylanases].

    Science.gov (United States)

    van Kampen, V; Merget, R; Brüning, T

    2004-02-01

    The exposure against enzyme dusts have long been known to cause occupational allergies. In the 1960s an increasing number of occupational allergies in the detergent industry were observed. In this context the high sensitization potential of enzyme dusts attracted attention. The present evaluation of literature data confirms that this is also true for xylanases. These frequently used industrial enzymes belong to the hemicellulases and are mostly of fungal origin. Several cases of specific airway sensitization caused by xylanases or other hemicellulases are verified by a number of case reports and cross sectional studies. As symptoms, results of skin prick tests, detection of specific IgE-antibodies and results of specific bronchoprovocation tests are consistent, an immunologic mechanism can be assumed.

  5. Ophthalmia neonatorum caused by Neisseria cinerea.

    OpenAIRE

    Bourbeau, P; Holla, V; Piemontese, S

    1990-01-01

    Neisseria cinerea is an organism that has only recently been implicated as a human pathogen. In this case, N. cinerea was identified as the cause of ophthalmia neonatorum (conjunctivitis) in a 2-day-old girl.

  6. Ophthalmia neonatorum caused by Neisseria cinerea.

    Science.gov (United States)

    Bourbeau, P; Holla, V; Piemontese, S

    1990-07-01

    Neisseria cinerea is an organism that has only recently been implicated as a human pathogen. In this case, N. cinerea was identified as the cause of ophthalmia neonatorum (conjunctivitis) in a 2-day-old girl.

  7. Knocking out Bcsas1 in Botrytis cinerea impacts growth, development, and secretion of extracellular proteins, which decreases virulence.

    Science.gov (United States)

    Zhang, Zhanquan; Qin, Guozheng; Li, Boqiang; Tian, Shiping

    2014-06-01

    Pathogenic fungi usually secrete a series of virulence factors to the extracellular environment to facilitate infection. Rab GTPases play a central role in the secretory pathway. To explore the function of Rab/GTPase in filamentous fungi, we knocked out a Rab/GTPase family gene, Bcsas1, in Botrytis cinerea, an aggressive fungal pathogen that infects more than 200 plant species. A detailed analysis was conducted on the virulence and the secretory capability of the mutants. The results indicated that knockout of Bcsas1 inhibited hyphal development and reduced sporulation of B. cinerea on potato dextrose agar plates resulting in reduced virulence on various fruit hosts. Knocking out the Bcsas1 gene led to an accumulation of transport vesicles at the hyphal tip, significantly reduced extracellular protein content, and lowered the activity of polygalacturonase and xylanase in the extracellular medium. However, mutation of Bcsas1 did not affect the expression of genes encoding polygalacturonase and xylanase, suggesting the secretion of these two family enzymes was suppressed in the mutant. Moreover, a comparative analysis of the secretome provided further evidence that the disruption of Bcsas1 in mutant strains significantly depressed the secretion of polysaccharide hydrolases and proteases. The results indicate that Bcsas1, the Rab8/SEC4-like gene, plays a crucial role in development, protein secretion, and virulence of B. cinerea.

  8. A Fusarium graminearum xylanase expressed during wheat infection is a necrotizing factor but is not essential for virulence.

    Science.gov (United States)

    Sella, Luca; Gazzetti, Katia; Faoro, Franco; Odorizzi, Silvana; D'Ovidio, Renato; Schäfer, Wilhelm; Favaron, Francesco

    2013-03-01

    Fusarium graminearum is the fungal pathogen mainly responsible for Fusarium head blight (FHB) of cereal crops, which attacks wheat spikes, reducing crop production and quality of grain by producing trichothecene mycotoxins. Several cytohistological studies showed that spike infection is associated with the production of cell wall degrading enzymes. Wheat tissue, as in other commelinoid monocot plants, is particularly rich in xylan which can be hydrolyzed by fungal endo-1,4-β-xylanase. The FG_03624 is one of the most expressed xylanase genes in wheat spikes 3 days after inoculation and was heterologously expressed in the yeast Pichia pastoris. The recombinant protein (22.7 kDa) possessed xylanase activity and induced cell death and hydrogen peroxide accumulation in wheat leaves infiltrated with 10 ng/μl or in wheat lemma surface treated with 20 ng/μl. This effect reflects that observed with other described fungal xylanases (from Trichoderma reesei, Trichoderma viride and Botrytis cinerea) with which the FG_03624 protein shares a stretch of amino acids reported as essential for elicitation of necrotic responses. Several F. graminearum mutants with the FG_03624 gene disrupted were obtained, and showed about 40% reduction of xylanase activity in comparison to the wild type when grown in culture with xylan as carbon source. However, they were fully virulent when assayed by single floret inoculation on wheat cvs. Bobwhite and Nandu. This is the first report of a xylanase able to induce hypersensitive-like symptoms on a monocot plant.

  9. Infection strategies of Botrytis cinerea

    NARCIS (Netherlands)

    Kan, van J.A.L.

    2005-01-01

    Botrytis cinerea is a ubiquitous filamentous fungal pathogen of a wide range of plant species. The fungus is able to infect all aerial parts of its host plants to a certain extent. Infection may cause enormous damage both during plant growth and in the post-harvest phase (during cold storage or tran

  10. Microbial xylanases: engineering, production and industrial applications.

    Science.gov (United States)

    Juturu, Veeresh; Wu, Jin Chuan

    2012-01-01

    Enzymatic depolymerization of hemicellulose to monomer sugars needs the synergistic action of multiple enzymes, among them endo-xylanases (EC 3.2.1.8) and β-xylosidases (EC 3.2.1.37) (collectively xylanases) play a vital role in depolymerizing xylan, the major component of hemicellulose. Recent developments in recombinant protein engineering have paved the way for engineering and expressing xylanases in both heterologous and homologous hosts. Functional expression of endo-xylanases has been successful in many hosts including bacteria, yeasts, fungi and plants with yeasts being the most promising expression systems. Functional expression of β-xylosidases is more challenging possibly due to their more complicated structures. The structures of endo-xylanases of glycoside hydrolase families 10 and 11 have been well elucidated. Family F/10 endo-xylanases are composed of a cellulose-binding domain and a catalytic domain connected by a linker peptide with a (β/α)8 fold TIM barrel. Family G/11 endo-xylanases have a β-jelly roll structure and are thought to be able to pass through the pores of hemicellulose network owing to their smaller molecular sizes. The structure of a β-D-xylosidase belonging to family 39 glycoside hydrolase has been elucidated as a tetramer with each monomer being composed of three distinct regions: a catalytic domain of the canonical (β/α)8--TIM barrel fold, a β-sandwich domain and a small α-helical domain with the enzyme active site that binds to D-xylooligomers being present on the upper side of the barrel. Glycosylation is generally considered as one of the most important post-translational modifications of xylanases, but a few examples showed functional expression of eukaryotic xylanases in bacteria. The optimal ratio of these synergistic enzymes is very important in improving hydrolysis efficiency and reducing enzyme dosage but has hardly been addressed in literature. Xylanases have been used in traditional fields such as food, feed

  11. Genetic variation and pathogenicity of Botrytis cinerea.

    NARCIS (Netherlands)

    Vlugt-Bergmans, van der C.J.B.

    1996-01-01

    Botrytis cinerea is a fungal pathogen of more than 200 hosts including a wide variety of economically important crops. Although many ecological and physiological studies on this destructive pathogen have been reported, not much is known about the molecular basis of the interaction of this pathogen w

  12. Cellulase-free xylanases from Bacillus and other microorganisms.

    Science.gov (United States)

    Subramaniyan, S; Prema, P

    2000-02-01

    Xylanases are used mainly in the pulp and paper industries for the pretreatment of Kraft pulp prior to bleaching to minimize use of chlorine, the conventional bleaching agent. This application has great potential as an environmentally safe method. Hydrolysis by xylanases of relocated and reprecipitated xylan on the surface of cellulose fibres formed during Kraft cooking facilitates the removal of lignin by increasing permeability to oxidising agents. Most of the xylanases reported in the literature contained significant cellulolytic activity, which make them less suitable for pulp and paper industries. The need for large quantities of xylanases which would be stable at higher temperatures and pH values and free of cellulase activity has necessitated a search for novel enzymes. We have isolated and characterised several xylanase-producing cultures, one of which (an alkalophilic Bacillus SSP-34) produced more than 100 IU ml(-1) of xylanase activity. The SSP-34 xylanases have optimum activity at 50 degrees C in a pH range 6-8, with only small amounts of cellulolytic activity (CMCase (0.4 IU ml(-1), pH 7), FPase (0.2 IU ml(-1), pH 7) and no activity at pH 9).

  13. The time of infection of apples by Botrytis cinerea Pers.

    OpenAIRE

    Hanna Bryk

    2013-01-01

    The time of infection of apple fruits by Botrytis cinerea Pers. was studied. Artificial inoculations with conidial suspensions of B. cinerea were done at different stages of fruit developmment (flowers, sets, fruits). In autumn the apples were harvested and stored at a temperature of 2°C for 4 months after which rotting caused by B. cinerea was evaluated. B. cinerea presence in the calyx of apples was checked throughout the growing season. This was done by plating flowers, apple and set calyc...

  14. Xylanase production by Trichoderma strains in solid substrate fermentation

    Institute of Scientific and Technical Information of China (English)

    Krisztina Kovacs; George Szakacs; Lew Christopher

    2004-01-01

    @@ The importance of microbial enzymes in pulp and paper manufacturing has grown significantly in the last two decades. Solid substrate fermentation (SSF) holds tremendous potential for the production of microbial enzymes of commercial interest. SSF can be of special interest in those processes where the crude fermented product (whole SSF culture, in situ enzyme) may be used directly as the enzyme source. Xylanase preparations practically free of cellulase activity are especially useful for biobleaching of crude cellulose pulps. Thirty-nine Trichoderma isolates have been screened in SSF for xylanase production on hardwood oxygen-delignified soda-aq pulp as carbon source and enzyme inducer.Xylanase activities varied between 0 and 2200 IU/g dry matter (DM) of initial substrate. In most instances, the simultaneously produced cellulase levels were below 1.0 Filter Paper Unit (FPU) /g DM. The xylanase to cellulase activity ratio varied in the range of 5 to 3500. The three most promising isolates (TUB F-1647, TUB F-1658 and TUB F-1684) yielded xylanase activity of 2040,1300 and 1500 IU/g DM xylanase, respectively, and 0.64, 0.43 and 0.43 FPU/g DM cellulase with a xylanase to cellulase activity ratio of 3200, 3000 and 3500, respectively. Wild strains F-1647, F-1658 and F-1684 were isolated from tree bark of Maldives, soils of Peru (last two), respectively.Medium optimization experiments to enhance the xylanase yield and to increase the xylanase to cellulase ratio have also been performed.

  15. Biotransformation of bioactive isocaryolanes by Botrytis cinerea.

    Science.gov (United States)

    Ascari, Jociani; Boaventura, Maria Amélia Diamantino; Takahashi, Jacqueline Aparecida; Durán-Patrón, Rosa; Hernández-Galán, Rosario; Macías-Sánchez, Antonio J; Collado, Isidro G

    2011-08-26

    The metabolism of the fungistatic agent (8R,9R)-8-methoxyisocaryolan-9-ol (4) by the fungus Botrytis cinerea has been investigated. Biotransformation of compound 4 yielded compounds 5 and 6-9. No dihydrobotrydial is observed after 4 days of incubation of compound 4. Separate biotransformation of (8R,9R)-isocaryolane-8,9-diol (5) yielded compounds 7-11. The evaluation of the fungistatic activity against B. cinerea of compounds 4, 5, and 6 is reported. (4R,8R,9R)-8-Methoxyisocaryolane-9,15-diol (6), a major metabolite of (8R,9R)-8-methoxyisocaryolan-9-ol (4), shows a much reduced biological activity when compared with the parent compound. Isocaryolane derivatives 6-11 are described for the first time.

  16. Tricuspid valve endocarditis due to Neisseria cinerea.

    Science.gov (United States)

    Benes, J; Dzupova, O; Krizova, P; Rozsypal, H

    2003-02-01

    Reported here is a case of infective endocarditis caused by the saprophytic species Neisseria cinerea. To the best of our knowledge, this etiology has not been documented in the medical literature previously. The patient was an intravenous drug addict who developed tricuspid endocarditis with lung embolism. The disease was cured after treatment with ampicillin/clavulanate that was changed to ceftriaxone after an embolic event.

  17. Three new metabolites from Botrytis cinerea.

    Science.gov (United States)

    Wang, Tian-Shan; Zhou, Jin-Yan; Tan, Hong

    2008-01-01

    Three new metabolites, gamma-abscisolactone (1), botrytisic acids A (3) and B (4) were isolated from the fermentation broth of Botrytis cinerea TB-3-H8. Their structures were elucidated on the basis of MS, IR, UV, and NMR spectroscopic data. Compound 2 was isolated from natural resource for the first time. The structure of 1 was further confirmed by single-crystal X-ray diffraction (CCDC-265897).

  18. Four new lactones from Botrytis cinerea.

    Science.gov (United States)

    Colmenares, Ana J; Durán-Patrón, Rosa M; Hernández-Galán, Rosario; Collado, Isidro G

    2002-11-01

    Four new lactones (1-4) have been isolated from Botrytis cinerea. Their structures were elucidated by interpretation of spectral data, mainly (1)H and (13)C NMR, including two-dimensional analysis (HOMOCOSY, HMQC, and HMBC). The phytotoxic activities of these new natural products have been evaluated. Compounds 1-3 were inactive, while 4 showed a phytotoxic effect when tested up to 250 ppm.

  19. XYLANASE PREBLEACHING ON NAOH-AQ WHEAT STRAW PULP

    Institute of Scientific and Technical Information of China (English)

    Caixia Li; Yongjun Deng; Ping Li; Guigan Fang; Shuchai Liu

    2004-01-01

    Before calcium hypochlorite bleaching (H) and chlorination,alkaline extraction, calcium hypochlorite three-stage-bleaching (CEH),we used a kind of hemicellulase, xylanase, to treat wheat straw pulp from Gaoyou Papermill.Xylanase pretreatment contained tow stages, the first stage was xylanase treatment, which was followed by alkaline extraction, the second stage. The xylanase could act on partial lignin and carbohydrate, chiefly xylan. The following alkaline extraction could dissolve something that could not be removed during the first stage. The result of pretreatment was to facilitate penetration of bleaching chemicals, to reduce effective chlorine consumption and to lower pollution loading of bleaching effluent. In the case of these two bleaching processes, the enzymatic pretreatment substantially enhanced the optical properties of the pulps. To calcium hypochlorite bleaching, strength properties of pulps were improved.

  20. XYLANASE PREBLEACHING ON NaOH-AQ WHEAT STRAW PULP

    Institute of Scientific and Technical Information of China (English)

    CaixiaLi; YongjunDeng; PingLi; GuiganFang; ShuchaiLiu

    2004-01-01

    Before calcium hypochlorite bleaching (H) and chlorination, alkaline extraction, calcium hypochlorite three-stage-bleaching (CEH),we used a kind of hemicellulase, xylanase, to treat wheat straw pulpfrom Gaoyou Papermill. Xylanase pretreatment contained tow stages, the first stage was xylanase treatment, which was followed by alkaline extraction, the second stage. The xylanase could act on partial lignin and carbohydrate, chiefly xylan. The following alkaline extraction could dissolve something that could not be removed during the first stage. The result of pretreatment was to facilitate penetration of bleaching chemicals, to reduce effective chlorine consumption and to lower pollution loading of bleaching effluent. In the case of these two bleaching processes, the enzymatic pretreatment substantially enhanced the optical properties of the pulps. To calcium hypochlorite bleaching, strength properties of pulps wereimproved.

  1. Xylanase supplementation to rye diets for growing pigs

    DEFF Research Database (Denmark)

    Nørgaard, Jan Værum; Pedersen, Trine Friis; Blaabjerg, Karoline;

    2016-01-01

    Supplementation of xylanase to pig diets can hydrolyze arabinoxylan (AX) into lower molecular weight compounds and thereby decrease the viscosity and improve nutrient utilization. Xylanase supplementation has, however, shown variable effects in diets containing wheat, rye, and combinations thereof....... Differences in animal age, enzyme source and dose, target substrate, and diet processing may explain this. The objective was to study the effect of increasing doses of endo-1,4-β-xylanase from Trichoderma reesei on apparent total tract digestibility (ATTD) of OM, starch, nonstarch polysaccharides (NSP), AX......, fat, N, and P of rye fed to growing pigs. Twenty-four 47-kg pigs were assigned to 4 diets containing 97.85% rye and 0, 4,000, 8,000, or 16,000 units xylanase/kg (as-fed basis). Pigs were placed in metabolic cages for 10 d: 5 d for adaption and 5 d for total but separate collection of feces and urine...

  2. A gapless genome sequence of the fungus Botrytis cinerea

    NARCIS (Netherlands)

    Kan, Van Jan A.L.; Stassen, Joost H.M.; Mosbach, Andreas; Lee, Van Der Theo A.J.; Faino, Luigi; Farmer, Andrew D.; Papasotiriou, Dimitrios G.; Zhou, Shiguo; Seidl, Michael F.; Cottam, Eleanor; Edel, Dominique; Hahn, Matthias; Schwartz, David C.; Dietrich, Robert A.; Widdison, Stephanie; Scalliet, Gabriel

    2016-01-01

    Following earlier incomplete and fragmented versions of a genome sequence for the grey mould Botrytis cinerea, a gapless, near-finished genome sequence for B. cinerea strain B05.10 is reported. The assembly comprised 18 chromosomes and was confirmed by an optical map and a genetic map based on ap

  3. Rapid development of xylanase assay conditions using Taguchi methodology.

    Science.gov (United States)

    Prasad Uday, Uma Shankar; Bandyopadhyay, Tarun Kanti; Bhunia, Biswanath

    2016-11-01

    The present investigation is mainly concerned with the rapid development of extracellular xylanase assay conditions by using Taguchi methodology. The extracellular xylanase was produced from Aspergillus niger (KP874102.1), a new strain isolated from a soil sample of the Baramura forest, Tripura West, India. Four physical parameters including temperature, pH, buffer concentration and incubation time were considered as key factors for xylanase activity and were optimized using Taguchi robust design methodology for enhanced xylanase activity. The main effect, interaction effects and optimal levels of the process factors were determined using signal-to-noise (S/N) ratio. The Taguchi method recommends the use of S/N ratio to measure quality characteristics. Based on analysis of the S/N ratio, optimal levels of the process factors were determined. Analysis of variance (ANOVA) was performed to evaluate statistically significant process factors. ANOVA results showed that temperature contributed the maximum impact (62.58%) on xylanase activity, followed by pH (22.69%), buffer concentration (9.55%) and incubation time (5.16%). Predicted results showed that enhanced xylanase activity (81.47%) can be achieved with pH 2, temperature 50°C, buffer concentration 50 Mm and incubation time 10 min.

  4. The time of infection of apples by Botrytis cinerea Pers.

    Directory of Open Access Journals (Sweden)

    Hanna Bryk

    2013-12-01

    Full Text Available The time of infection of apple fruits by Botrytis cinerea Pers. was studied. Artificial inoculations with conidial suspensions of B. cinerea were done at different stages of fruit developmment (flowers, sets, fruits. In autumn the apples were harvested and stored at a temperature of 2°C for 4 months after which rotting caused by B. cinerea was evaluated. B. cinerea presence in the calyx of apples was checked throughout the growing season. This was done by plating flowers, apple and set calyces on PDA medium. Latent infection of apples by B. cinerea was found. The infection took place in the orchard and the pathogen survived latently in the calyx; disease symptoms appeared in storage. Infection is possible during the whole vegetative season, but the most important time is flowering and just before or just after harvest of apples.

  5. Cloning of xylanase genes from Ruminococcus albus and chromosome mapping of Fibrobacter succinogenes

    DEFF Research Database (Denmark)

    Nagamine, T.; Aminov, Rustam; Ogata, K.

    1997-01-01

    The characterization of the xylanase genes of Ruminococcus albus and the genomic organization of Fibrobacter succinogenes are described.......The characterization of the xylanase genes of Ruminococcus albus and the genomic organization of Fibrobacter succinogenes are described....

  6. Purification and characterization of thermoalkalophilic xylanase isolated from the Enterobacter sp. MTCC 5112

    Digital Repository Service at National Institute of Oceanography (India)

    Khandeparker, R.; Bhosle, N.B.

    Thermoalkalophilic Enterobacter sp MTCC 5112 was isolated from a sediment sample collected from the Mandovi estuary, west coat of India. This culture produced extracellular xylanase. The xylanase enzyme was isolated by ammonium sulfate (80...

  7. Ozone injury increases infection of geranium leaves by Botrytis cinerea

    Energy Technology Data Exchange (ETDEWEB)

    Manning, W.J.; Feder, W.A.; Perkins, I.

    1970-04-01

    Detached and attached, inoculated and noninoculated, ozone-injured and noninjured leaves from the lower, middle, and terminal regions of plants of geranium cultivars Enchantress and White Mountain were observed for infection by Botrytis cinerea. Previous exposure to ozone did not appreciably influence the susceptibility of leaves of either geranium cultivar to infection by B. cinerea, unless there was visible ozone injury. Ozone-injured, necrotic tissues on older attached and detached geranium leaves of both cultivars served as infection courts for B. cinerea. 14 references, 1 table.

  8. Ozone injury and infection of potato leaves by Botrytis cinerea

    Energy Technology Data Exchange (ETDEWEB)

    Manning, W.J.; Feder, W.A.; Perkins, I.; Glickman, M.

    1969-09-01

    Symptoms of ozone injury were observed on older leaves of potato cultivars Norland and Katahdin under experimental conditions. This symptom expression closely resembled flecks observed on potato leaves also blighted by Botrytis cinerea in the field. Inoculation of ozone-injured and noninjured potato leaves with B. cinerea showed that infection was more rapid and disease development more severe on ozone-injured leaves. Infection was frequently observed to originate in ozone-injured leaf areas. Ozone injury, under experimental conditions, appeared to increase the susceptibility of potato leaves to infection by B. cinerea. 6 references.

  9. ENZYMATIC DEINKING OF OLD NEWSPRINT WITH CELLULASES AND XYLANASE

    Institute of Scientific and Technical Information of China (English)

    Shoujuan Wang; Menghua Qin; Yingjuan Fu; Zhiyong Shao

    2004-01-01

    The enzymatic deinking and fiber modification of old newsprint (ONP) with several cellulases and xylanase were investigated and the suitable enzyme candidates were selected for ONP deinking in this paper. The results demonstrated that the cellulases and hemicellulases could significantly improve the deinking efficiency and fiber modification.Moreover, the synergistic effects of Novozym342 and xylanase (HC) can further enhance the deinking performance, reduce the dirt count and improve the brightness of resulting pulp. Additionally, compared to deinked pulps, obtained from conventional chemical materials, enzymatically deinked pulps had better bleachability, and the brightness of the bleached pulp reached 59.1% ISO, 9% ISO higher than the unbleached pulp.

  10. ENZYMATIC DEINKING OF OLD NEWSPRINT WITH CELLULASES AND XYLANASE

    Institute of Scientific and Technical Information of China (English)

    ShoujuanWang; MenghuaQin; YingjuanFu; ZhiyongShao

    2004-01-01

    The enzymatic deinking and fiber modification of old newsprint (ONP) with several cellulases and xylanase were investigated and the suitable enzyme candidates were selected for ONP deinking in this paper. The results demonstrated that the cellulases and hemicellulases could significantly improve the deinking efficiency and fiber modification. Moreover, the synergistic eflbcts of Novozym342 and xylanase (HC) can further enhance the deinking performance, reduce the dirt count and improve the brightness of resulting pulp. Additionally, compared to deinked pulps, obtained from conventional chemical materials, enzymatically deinked pulps had better bleachability, and the brightness of the bleached pulp reached 59.1% ISO, 9% ISO higher than the unbleached pulp.

  11. Proteomic analysis of ripening tomato fruit infected by Botrytis cinerea.

    Science.gov (United States)

    Shah, Punit; Powell, Ann L T; Orlando, Ron; Bergmann, Carl; Gutierrez-Sanchez, Gerardo

    2012-04-06

    Botrytis cinerea, a model necrotrophic fungal pathogen that causes gray mold as it infects different organs on more than 200 plant species, is a significant contributor to postharvest rot in fresh fruit and vegetables, including tomatoes. By describing host and pathogen proteomes simultaneously in infected tissues, the plant proteins that provide resistance and allow susceptibility and the pathogen proteins that promote colonization and facilitate quiescence can be identified. This study characterizes fruit and fungal proteins solubilized in the B. cinerea-tomato interaction using shotgun proteomics. Mature green, red ripe wild type and ripening inhibited (rin) mutant tomato fruit were infected with B. cinerea B05.10, and the fruit and fungal proteomes were identified concurrently 3 days postinfection. One hundred eighty-six tomato proteins were identified in common among red ripe and red ripe-equivalent ripening inhibited (rin) mutant tomato fruit infected by B. cinerea. However, the limited infections by B. cinerea of mature green wild type fruit resulted in 25 and 33% fewer defense-related tomato proteins than in red and rin fruit, respectively. In contrast, the ripening stage of genotype of the fruit infected did not affect the secreted proteomes of B. cinerea. The composition of the collected proteins populations and the putative functions of the identified proteins argue for their role in plant-pathogen interactions.

  12. Molecular cloning and characterization of multidomain xylanase from manure library

    Science.gov (United States)

    The gene (manf-x10) encoding xylanase from an environmental genomic DNA library was cloned and expressed in Escherichia coli. The encoded enzyme was predicted to be 467 amino acids with a molecular mass of 50.3 kD. The recombinant ManF-X10 was purified by HisTrap affinity column and showed activit...

  13. A multipurpose immobilized biocatalyst with pectinase, xylanase and cellulase activities

    Directory of Open Access Journals (Sweden)

    Gupta Munishwar

    2007-06-01

    Full Text Available Abstract Background The use of immobilized enzymes for catalyzing various biotransformations is now a widely used approach. In recent years, cross-linked enzyme aggregates (CLEAs have emerged as a novel and versatile biocatalyst design. The present work deals with the preparation of a CLEA from a commercial preparation, Pectinex™ Ultra SP-L, which contains pectinase, xylanase and cellulase activities. The CLEA obtained could be used for any of the enzyme activities. The CLEA was characterized in terms of kinetic parameters, thermal stability and reusability in the context of all the three enzyme activities. Results Complete precipitation of the three enzyme activities was obtained with n-propanol. When resulting precipitates were subjected to cross-linking with 5 mM glutaraldehyde, the three activities initially present (pectinase, xylanase and cellulase were completely retained after cross-linking. The Vmax/Km values were increased from 11, 75 and 16 to 14, 80 and 19 in case of pectinase, xylanase and cellulase activities respectively. The thermal stability was studied at 50°C, 60°C and 70°C for pectinase, xylanase and cellulase respectively. Half-lives were improved from 17, 22 and 32 minutes to 180, 82 and 91 minutes for pectinase, xylanase and cellulase respectively. All three of the enzymes in CLEA could be reused three times without any loss of activity. Conclusion A single multipurpose biocatalyst has been designed which can be used for carrying out three different and independent reactions; 1 hydrolysis of pectin, 2 hydrolysis of xylan and 3 hydrolysis of cellulose. The preparation is more stable at higher temperatures as compared to the free enzymes.

  14. The xylanase inhibitor TAXI-III counteracts the necrotic activity of a Fusarium graminearum xylanase in vitro and in durum wheat transgenic plants.

    Science.gov (United States)

    Moscetti, Ilaria; Faoro, Franco; Moro, Stefano; Sabbadin, Davide; Sella, Luca; Favaron, Francesco; D'Ovidio, Renato

    2015-08-01

    The xylanase inhibitor TAXI-III has been proven to delay Fusarium head blight (FHB) symptoms caused by Fusarium graminearum in transgenic durum wheat plants. To elucidate the molecular mechanism underlying the capacity of the TAXI-III transgenic plants to limit FHB symptoms, we treated wheat tissues with the xylanase FGSG_03624, hitherto shown to induce cell death and hydrogen peroxide accumulation. Experiments performed on lemmas of flowering wheat spikes and wheat cell suspension cultures demonstrated that pre-incubation of xylanase FGSG_03624 with TAXI-III significantly decreased cell death. Most interestingly, a reduced cell death relative to control non-transgenic plants was also obtained by treating, with the same xylanase, lemmas of TAXI-III transgenic plants. Molecular modelling studies predicted an interaction between the TAXI-III residue H395 and residues E122 and E214 belonging to the active site of xylanase FGSG_03624. These results provide, for the first time, clear indications in vitro and in planta that a xylanase inhibitor can prevent the necrotic activity of a xylanase, and suggest that the reduced FHB symptoms on transgenic TAXI-III plants may be a result not only of the direct inhibition of xylanase activity secreted by the pathogen, but also of the capacity of TAXI-III to avoid host cell death.

  15. Association of Neisseria cinerea with ocular infections in paediatric patients.

    Science.gov (United States)

    Dolter, J; Wong, J; Janda, J M

    1998-01-01

    Twenty-two strains of Neisseria cinerea were recovered from paediatric patients over a 7-year period and forwarded to the Microbial Diseases Laboratory for biochemical identification and/or confirmation. Eighteen of these 22 strains (82%) were recovered from the eyes of very young children ( 50% occurring during the neonatal period. The majority of eye isolates were involved in a variety of ocular infections including orbital cellulitis, conjunctivitis, and eye discharge (most common); in four of the 13 instances (31%) where laboratory data was available, Neisseria cinerea was recovered in pure culture. Neisseria cinerea isolates were often submitted to the Microbial Diseases Laboratory as possible 'N. gonorrhoeae' or 'Neisseria species' due to problems resulting from the use of commercial assays or unfamiliarity with the organism. These observations indicate that N. cinerea can produce eye infections in very young children, who presumably acquire this organism vertically from the mother during birth. Accurate identification of N. cinerea in such infants can preclude the social trauma and possible legal ramifications which can initially result from its misidentification as N. gonorrhoeae.

  16. Safety evaluation of a xylanase expressed in Bacillus subtilis.

    Science.gov (United States)

    Harbak, L; Thygesen, H V

    2002-01-01

    A programme of studies was conducted to establish the safety of a xylanase expressed in a self-cloned strain of Bacillus subtilis to be used as a processing aid in the baking industry. To assess acute and subchronic oral toxicity, rat feeding studies were conducted. In addition, the potential of the enzyme to cause mutagenicity and chromosomal aberrations was assessed in microbial and tissue culture in vitro studies. Acute and subchronic oral toxicity was not detected at the highest dose recommended by OECD guidelines. There was no evidence of mutagenic potential or chromosomal aberrations. Furthermore, the organism used for production of the xylanase is already accepted as safe by several major national regulatory agencies.

  17. Characterization and biotechnological application of recombinant xylanases from Aspergillus nidulans

    NARCIS (Netherlands)

    Maitan-Alfenas, Gabriela P; Oliveira, Mariana B; Nagem, Ronaldo A P; de Vries, Ronald P; Guimarães, Valéria M

    2016-01-01

    Two xylanases from Aspergillus nidulans, XlnB and XlnC, were expressed in Pichia pastoris, purified and characterized. XlnB and XlnC achieved maximal activities at 60°C and pH 7.5 and at 50°C and pH 6.0, respectively. XlnB showed to be very thermostable by maintaining 50% of its original activity af

  18. Bifunctional xylanases and their potential use in biotechnology

    Digital Repository Service at National Institute of Oceanography (India)

    Khandeparker, R.; Numan, M.Th.

    is half as sweet as sucrose, can be applicable to foods as a sweetener that is capable of improving diabetic symptoms [37]. Concluding remarks This review provides the information on most of the aspects of bifunctional enzyme with special reference... of the bifunctional xylanases it is necessary in future to utilize such hybrid protein as an alternative to expensive and polluting chemical treatments or to improve already existing enzymatic processes for utilization of veg- etal by-products in the agro...

  19. Fusarium graminearum produces different xylanases causing host cell death that is prevented by the xylanase inhibitors XIP-I and TAXI-III in wheat.

    Science.gov (United States)

    Tundo, Silvio; Moscetti, Ilaria; Faoro, Franco; Lafond, Mickaël; Giardina, Thierry; Favaron, Francesco; Sella, Luca; D'Ovidio, Renato

    2015-11-01

    To shed light on the role of Xylanase Inhibitors (XIs) during Fusarium graminearum infection, we first demonstrated that three out of four F. graminearum xylanases, in addition to their xylan degrading activity, have also the capacity to cause host cell death both in cell suspensions and wheat spike tissue. Subsequently, we demonstrated that TAXI-III and XIP-I prevented both the enzyme and host cell death activities of F. graminearum xylanases. In particular, we showed that the enzymatic inhibition by TAXI-III and XIP-I was competitive and only FGSG_11487 escaped inhibition. The finding that TAXI-III and XIP-I prevented cell death activity of heat inactivated xylanases and that XIP-I precluded the cell death activity of FGSG_11487 - even if XIP-I does not inhibit its enzyme activity - suggests that the catalytic and the cell death activities are separated features of these xylanases. Finally, the efficacy of TAXI-III or XIP-I to prevent host cell death caused by xylanases was confirmed in transgenic plants expressing separately these inhibitors, suggesting that the XIs could limit F. graminearum infection via direct inhibition of xylanase activity and/or by preventing host cell death.

  20. Isolation, purification and characterization of xylanase produced by Arthrobacter sp. MTCC 5214 when grown in solid-state fermentation

    Digital Repository Service at National Institute of Oceanography (India)

    Khandeparker, R.; Bhosle, N.B.

    Thermoalkalophilic Arthrobacter sp. produced extracellular xylanase, when wheat bran, rice husk, rice bran and bagassae were used as carbon source under solid-state fermentation (SSF). The xylanase enzyme was isolated by ammonium sulfate (80...

  1. Botrytis cinerea: the cause of grey mould disease

    NARCIS (Netherlands)

    Williamson, B.; Tudzynski, B.; Tudzynski, P.; Kan, van J.A.L.

    2007-01-01

    Introduction: Botrytis cinerea (teleomorph: Botryotinia fuckeliana) is an airborne plant pathogen with a necrotrophic lifestyle attacking over 200 crop hosts worldwide. Although there are fungicides for its control, many classes of fungicides have failed due to its genetic plasticity. It has become

  2. ABC transporters from Botrytis cinerea in biotic and abiotic interactions

    NARCIS (Netherlands)

    Schoonbeek, H.

    2004-01-01

    Botrytis cinereais the causal agent of grey mould disease on a wide variety of crop plants. It is relatively insensitive to natural and synthetic fungitoxic compounds. This thesis describes how ABC (ATP-binding cassette) transporters contribute to protection by actively secre

  3. ABC transporters van Botrytis cinerea in biotische en abiotische interacties

    NARCIS (Netherlands)

    Schoonbeek, H.

    2005-01-01

    Op 29 november 2004 promoveerde Henk-jan Schoonbeek aan Wageningen Universiteit op het proefschrift getiteld 'ABC transporters from Botrytis cinerea in biotic and abiotic interactions'. Promotor was Prof. dr. ir. P.J.G.M. de Wit en co-promotor was dr.ir. M.A. de Waard, leerstoelgroep Fytopathologie,

  4. Optimizing chemically induced resistance in tomato against Botrytis cinerea

    DEFF Research Database (Denmark)

    Luna, Estrella; Beardon, Emily G; Ravnskov, Sabine

    2016-01-01

    resistance in tomato against Botrytis cinerea. In addition, we have studied non-target effects on plant growth and root colonization by arbuscular mycorrhizal fungi (AMF). Germinating seeds for one week in BABA- or JA-containing solutions promoted seed germination efficiency, did not affect plant growth...

  5. Bacteremia due to Neisseria cinerea: report of two cases.

    Science.gov (United States)

    Southern, P M; Kutscher, A E

    1987-06-01

    We report two cases of bacteremia due to Neisseria cinerea. One was a 2.5-yr-old boy with otitis media and pneumonia, who responded to treatment with amoxicillin. The other was a 47-yr-old man with underlying ethanol abuse who developed severe polymicrobial sepsis due to apparent intraabdominal disease. This man died despite extensive antimicrobial therapy.

  6. Changes in Botrytis cinerea Conidia Caused by Berberis vulgaris Extract

    Directory of Open Access Journals (Sweden)

    Marcel PARVU

    2010-12-01

    Full Text Available Testing plant extracts for controlling fungal diseases is a main biocontrol method. More interesting is to see what happens to the fungus treated with the plant extract. Therefore, the aim of the study was to evaluate the antifungal activity of Berberis vulgaris extract on Botrytis cinerea and to examine the ultrastructural changes in B. cinerea conidia caused by the minimum inhibitory concentration (MIC, using SEM and TEM. The antifungal activity of B. vulgaris bark extract was investigated using agar dilution method, and compared to that of berberine. Fluconazole was used as the positive antimycotic control. It was found that (1 B. vulgaris bark extract had significant antifungal activity against B. cinerea, and its effect was stronger than that of pure berberine. It was also noted that (2B. vulgaris MIC caused severe structural changes of the conidia, comparable with berberine MIC effect; therefore (3 B. vulgaris bark extract might be recommended to be tested as a biocontrol agent against B. cinerea.

  7. The D-galacturonic acid catabolic pathway in Botrytis cinerea.

    Science.gov (United States)

    Zhang, Lisha; Thiewes, Harry; van Kan, Jan A L

    2011-10-01

    D-galacturonic acid is the most abundant component of pectin, one of the major polysaccharide constituents of plant cell walls. Galacturonic acid potentially is an important carbon source for microorganisms living on (decaying) plant material. A catabolic pathway was proposed in filamentous fungi, comprising three enzymatic steps, involving D-galacturonate reductase, L-galactonate dehydratase, and 2-keto-3-deoxy-L-galactonate aldolase. We describe the functional, biochemical and genetic characterization of the entire D-galacturonate-specific catabolic pathway in the plant pathogenic fungus Botrytis cinerea. The B. cinerea genome contains two non-homologous galacturonate reductase genes (Bcgar1 and Bcgar2), a galactonate dehydratase gene (Bclgd1), and a 2-keto-3-deoxy-L-galactonate aldolase gene (Bclga1). Their expression levels were highly induced in cultures containing GalA, pectate, or pectin as the sole carbon source. The four proteins were expressed in Escherichia coli and their enzymatic activity was characterized. Targeted gene replacement of all four genes in B. cinerea, either separately or in combinations, yielded mutants that were affected in growth on D-galacturonic acid, pectate, or pectin as the sole carbon source. In Aspergillus nidulans and A. niger, the first catabolic conversion only involves the Bcgar2 ortholog, while in Hypocrea jecorina, it only involves the Bcgar1 ortholog. In B. cinerea, however, BcGAR1 and BcGAR2 jointly contribute to the first step of the catabolic pathway, albeit to different extent. The virulence of all B. cinerea mutants in the D-galacturonic acid catabolic pathway on tomato leaves, apple fruit and bell peppers was unaltered.

  8. Autohydrolysis of plant xylans by apoplastic expression of thermophilic bacterial endo-xylanases

    DEFF Research Database (Denmark)

    Borkhardt, Bernhard; Harholt, Jesper; Ulvskov, Peter Bjarne

    2010-01-01

    The genes encoding the two endo-xylanases XynA and XynB from the thermophilic bacterium Dictyoglomus thermophilum were codon optimized for expression in plants. Both xylanases were designed to be constitutively expressed under the control of the CaMV 35S promoter and targeted to the apoplast. Tra...

  9. Baker's asthma due to the enzyme xylanase -- a new occupational allergen.

    Science.gov (United States)

    Baur, X; Sander, I; Posch, A; Raulf-Heimsoth, M

    1998-12-01

    The asthmatic baker showed IgE-mediated sensitization to xylanase of Aspergillus niger used as a baking additive. Inhalative challenge with approximately 0.5 microg of the enzyme resulted in an immediate-type asthmatic reaction. This case, as well as a preliminary screening of symptomatic bakers, shows that xylanase is a further relevant type I-sensitizer in the baking industry.

  10. A novel halotolerant xylanase from marine isolate Bacillus subtilis cho40: gene cloning and sequencing

    Digital Repository Service at National Institute of Oceanography (India)

    Khandeparker, R.; Verma, P.; Deobagkar, D

    induced (140%) when pre-incubated with 0.5 M NaCl for 4 h. The xylanase gene, xyn40, from marine bacterium B. subtilis cho40 was cloned, and expressed in Escherichia coli. The xylanase gene was 645 bp long and had a 215 amino acid ORF protein with a...

  11. A novel thermoalkaliphilic xylanase from Gordonia sp. is salt, solvent and surfactant tolerant.

    Science.gov (United States)

    Kashyap, Radhika; Monika; Subudhi, Enketeswara

    2014-12-01

    Two aerobic bacterial consortia namely Con T and Con R were developed by enrichment technique from termite gut and cow dung respectively, using xylan as a sole carbon source. Molecular characterization of Con R based on 16S rRNA sequence analysis showed the presence of Pannonibacter sp. R-3 and Pseudoxanthomas sp. R-5. On the other hand, Con T showed the presence of Pseudoxanthomas sp. T-5, Cellulosimicrobium sp. T-21, and Gordonia sp. T-30. Being the maximum xylanase producer among the five isolates and being a novel xylanase producing bacterial genus, Gordonia sp. T-30 was selected. Xylanase produced by Gordonia sp. T-30 showed optimum activity at 60 °C and pH 9. Xylanase was 95% stable for 120 min at pH 9.0 and 98% stable at 60 °C for 90 min. Xylanase activity was stimulated in the presence of organic solvents such as petroleum ether, acetone, diethyl ether, n-hexane, and benzene. Detergent like cetyltrimethylammonium bromide and presence of NaCl also accelerated the xylanase function. Comparative evaluation was studied between sterilized and non-sterilized solid fermentation to produce xylanase by Gordonia sp. T-30 using various agricultural residues as growth substrate in cost effective manner. Industrially important features endowed by this xylanase make it a very promising candidate for food, feed, and fuel industry.

  12. Performance and morphometry of the intestinal mucosa of laying hens fed diets containing xylanase

    Directory of Open Access Journals (Sweden)

    KMR de Souza

    2014-09-01

    Full Text Available The objective of this study was to evaluate the effect of dietary energy level reduction and xylanase inclusion on the performance and on intestinal mucosa morphometry of two- to six-week-old laying hens. In total, 400 Hy-line W36 laying hens were distributed according to a completely randomized design in 2 x 2 factorial arrangement (energy level x inclusion of xylanase, totaling four treatments with 10 replicates of 10 birds per experimental unit. The following treatments were evaluated: positive control (balanced diet; positive control + xylanase; negative control (diet with of 100 kcal ME reduction /kg; negative control + xylanase. Body weight, weight gain, feed conversion ratio, uniformity and livability were not influenced by diets with metabolizable energy reduction and xylanase inclusion; however, the addition of xylanase to the diets resulted in shallower crypts depth and greater villus:crypt ratio in the ileum. The energy reduction of the diet associated with the supplementation of xylanase did not influence performance, but increased the feed intake of 2- to 6-week-old laying hens and increased villus height in the ileum of 6-wk-old hens. Xylanase reduces crypt depth in the ileum of 6-week-old hens.

  13. Benefits from additives and xylanase during enzymatic hydrolysis of bamboo shoot and mature bamboo.

    Science.gov (United States)

    Li, Kena; Wang, Xiao; Wang, Jingfeng; Zhang, Junhua

    2015-09-01

    Effects of additives (BSA, PEG 6000, and Tween 80) on enzymatic hydrolysis of bamboo shoot and mature bamboo fractions (bamboo green, bamboo timber, bamboo yellow, bamboo node, and bamboo branches) by cellulases and/or xylanase were evaluated. The addition of additives was comparable to the increase of cellulase loadings in the conversion of cellulose and xylan in bamboo fractions. Supplementation of xylanase (1 mg/g DM) with cellulases (10 FPU/g DM) in the hydrolysis of bamboo fractions was more efficient than addition of additives in the production of glucose and xylose. Moreover, addition of additives could further increase the glucose release from different bamboo fractions by cellulases and xylanase. Bamboo green exhibited the lowest hydrolyzability. Almost all of the polysaccharides in pretreated bamboo shoot fractions were hydrolyzed by cellulases with the addition of additives or xylanase. Additives and xylanase showed great potential for reducing cellulase requirement in the hydrolysis of bamboo.

  14. [Cellulase and xylanase activity of phytopathogenic and endophytic fungal strains of Alternaria alternata (Fr.) Keissler].

    Science.gov (United States)

    Kurchenko, I M; Sokolova, O V; Zhdanova, N M; Iarynchyn, A M; Iovenko, O M

    2008-01-01

    A comparative analysis of cellulase and xylanase activity of 25 fungal strains of phytopathogenic and endophytic Alternaria alternata had been realized for the first time using the qualitative reactions. The rate of their linear growth on the media with carboxymethylcellulose or xylane had been studied. The cellulase and xylanase activities clearly depended on the distinct strain. The absence of distinct dependence of cellulase and xylanase activities on the species and organs of host plants was demonstrated. The majority of investigated strains of A. alternata did not possess a cellulase activity or the latter was low, but as a whole the phytopathogenic strains were more active than endophytic ones. Xylanase activity was considerable for the fungal strains of all trophyc groups. It was shown that the level of xylanase activity cannot become a biochemical marker of the A. alternata isolate pathogenicity.

  15. Production of UC-labeled gas in BACTEC Neisseria Differentiation kits by Neisseria cinerea

    Energy Technology Data Exchange (ETDEWEB)

    Boyce, J.M.; Mitchell, E.B. Jr.; Knapp, J.S.; Buttke, T.M.

    1985-09-01

    Six strains of Neisseria cinerea were tested in BACTEC Neisseria Differentiation kits (Johnston Laboratories, Inc., Towson, Md.), and all yielded positive glucose growth indices and negative maltose and fructose growth indices. These results were similar to those achieved with Neisseria gonorrhoeae. However, most of the N. cinerea isolates tested yielded 3-h glucose growth indices that were lower than those obtained with gonococci. UC-labeled gas was produced significantly faster by N. gonorrhoeae than by N. cinerea. Additional studies suggested that the UC-labeled gas produced by N. cinerea was carbon dioxide. N. cinerea strains were similar to Branhamella catarrhalis strains because both species failed to produce detectable acid from glucose, maltose, sucrose, fructose, and lactose in cysteine-tryptic agar media. However, in contrast to N. cinerea strains, B. catarrhalis strains did not metabolize glucose in BACTEC Neisseria Differentiation kits.

  16. Production of 14C-labeled gas in BACTEC Neisseria Differentiation kits by Neisseria cinerea.

    Science.gov (United States)

    Boyce, J M; Mitchell, E B; Knapp, J S; Buttke, T M

    1985-09-01

    Six strains of Neisseria cinerea were tested in BACTEC Neisseria Differentiation kits (Johnston Laboratories, Inc., Towson, Md.), and all yielded positive glucose growth indices and negative maltose and fructose growth indices. These results were similar to those achieved with Neisseria gonorrhoeae. However, most of the N. cinerea isolates tested yielded 3-h glucose growth indices that were lower than those obtained with gonococci. 14C-labeled gas was produced significantly faster (P less than 0.02) by N. gonorrhoeae than by N. cinerea. Additional studies suggested that the 14C-labeled gas produced by N. cinerea was carbon dioxide. N. cinerea strains were similar to Branhamella catarrhalis strains because both species failed to produce detectable acid from glucose, maltose, sucrose, fructose, and lactose in cysteine-tryptic agar media. However, in contrast to N. cinerea strains, B. catarrhalis strains did not metabolize glucose in BACTEC Neisseria Differentiation kits.

  17. Partial purification and characterization of xylanase produced by Penicillium expansum

    Directory of Open Access Journals (Sweden)

    André Luiz de Souza Querido

    2006-05-01

    Full Text Available An extracellular xylanase was found to be the major protein in the filtrate culture of Penicillium expansum when grown on 0.3 % wheat bran, which showed no xylanase multiplicity. The enzime was partial purified by.ammonium sulfate fractioning, molecular exclusion chromatography, ultrafiltration and anion exchange chromatography. The protein eluation profile showed only one form of xylanase that was partially characterized. The activity of purified xylanase was optimal at pH 5.5 and 40 ºC. The enzyme was stable at pH between 5.5 and 6.5 and temperatures between 20-40 ºC. The enzyme showed a Km of 3.03 mM and Vmax of 0.027 mumol min-1 mug -1 of protein. The enzymatic activity was increased 31 % by Mg2+ and 28 % by Al3+.Uma xilanase extracelular foi encontrada como a principal proteína na cultura filtrada de Penicillium expansum quando cultivado em farelo de trigo 0,3 %, a qual não mostrou multiplicidade. A enzima foi parcialmente purificada por fracionamento com sulfato de amônia, cromatografia de exclusão molecular, ultrafiltração e cromatografia de troca aniônica. O perfil de eluição das proteínas mostrou uma única forma de xilanase, sendo esta parcialmente caracterizada. A atividade da xilanase purificada foi ótima em pH 5.5 e à temperatura de 40 ºC. A enzima foi estável em pH entre 5,5 e 6,5 e à temperatura entre 20-40ºC. A enzima apresentou Km de 3,03 mM e Vmax de 0,027 mimol min-1 mig-1 de proteína. A atividade enzimática foi aumentada 31 % por Mg+2 e 28 % por Al+3.

  18. The effect of harpin on shelf life of peppers inoculated with Botrytis cinerea

    OpenAIRE

    TEZCAN, Himmet; Akbudak, Nuray; Akbudak, Bulent

    2011-01-01

    The preservation methods as an alternative to chemical control to prevent postharvest quality losses of peppers were examined. The efficacy of harpin treatments on peppers (Capsicum annuum L. cvs. ‘Demre’, ‘Yalova Charleston’ and ‘Sari Sivri’) was tested in the same conditions in two different years. Peppers grown in greenhouse were applied with four treatments consisting of harpin, Botrytis cinerea, harpin+B. cinerea and control. The harpin in B. cinerea treatments reduced the percentage of ...

  19. Some biochemical reactions of strawberry plants to infection with Botrytis cinerea and salicylic acid treatment

    Directory of Open Access Journals (Sweden)

    Urszula Małolepsza

    2013-12-01

    Full Text Available The reactions of strawberry plants to infection with B. cinerea and treatment with salicylic acid has been studied. Infection of leaves with B. cinerea resulted in early increases in active oxygen species generation, superoxide dismutase and peroxidase activities and phenolic compounds content. Some increases of the above reactions were noticed in plants treated with salicylic acid but not in the plants treated with SA and then later infected with B. cinerea.

  20. Prevalence and persistence of Neisseria cinerea and other Neisseria spp. in adults.

    OpenAIRE

    Knapp, J S; Hook, E W

    1988-01-01

    Neisseria cinerea is a commensal Neisseria sp. which was first described in 1906 but was subsequently misclassified as a subtype of Branhamella catarrhalis. N. cinerea resembles Neisseria gonorrhoeae in both cultural and biochemical characteristics and, thus, may also have been misidentified as N. gonorrhoeae. Of 202 patients whose oropharynges were colonized by Neisseria spp., N. cinerea was isolated in 57 (28.2%) patients, including 25 (30.1%) of 83 women, 22 (23.9%) of 92 heterosexual men,...

  1. Antagonism of Trichoderma harzianum ETS 323 on Botrytis cinerea mycelium in culture conditions.

    Science.gov (United States)

    Cheng, Chi-Hua; Yang, Chia-Ann; Peng, Kou-Cheng

    2012-11-01

    ABSTRACT Previous studies have shown that the extracellular proteins of Trichoderma harzianum ETS 323 grown in the presence of deactivated Botrytis cinerea in culture include a putative l-amino acid oxidase and have suggested the involvement of this enzyme in the antagonistic mechanism. Here, we hypothesized that the mycoparasitic process of Trichoderma spp. against B. cinerea involves two steps; that is, an initial hyphal coiling stage and a subsequent hyphal coiling stage, with different coiling rates. The two-step antagonism of T. harzianum ETS 323 against B. cinerea during the mycoparasitic process in culture was evaluated using a biexponential equation. In addition, an l-amino acid oxidase (Th-l-AAO) was identified from T. harzianum ETS 323. The secretion of Th-l-AAO was increased when T. harzianum ETS 323 was grown with deactivated hyphae of B. cinerea. Moreover, in vitro assays indicated that Th-l-AAO effectively inhibited B. cinerea hyphal growth, caused cytosolic vacuolization in the hyphae, and led to hyphal lysis. Th-l-AAO also showed disease control against the development of B. cinerea on postharvest apple fruit and tobacco leaves. Furthermore, an apoptosis-like response, including the generation of reactive oxygen species, was observed in B. cinerea after treatment with Th-l-AAO, suggesting that Th-l-AAO triggers programmed cell death in B. cinerea. This may be associated with the two-step antagonism of T. harzianum ETS 323 against B. cinerea.

  2. Impact of essential oils on mycelial growth of Botrytis cinerea

    Directory of Open Access Journals (Sweden)

    Slavko Grgić

    2016-12-01

    Full Text Available The aim of this study was to determine the effect of 22 essential oils (anise, thyme, cumin, peppermint, lavender, sage, lemon balm, rosemary, myrtle, cinnamon leaf, basil, white pine, eucalyptus, cedar, bergamot, mandarin, cypress, patchouli, ginger, bitter orange, sandalwood, camphor on the growth of gray mold fungus Botrytis cinerea. The experiment was performed in vitro on PDA medium in 2 repetitions. Oils were applied in three amounts (3, 5 and 7 μl, and the mycelial growth was measured after three and nine days of incubation. All oils, except oils of bitter orange, sandalwood and camphor, have shown a certain antifungal activity. Compared to the water control, thyme and anise oil have shown the best antifungal activity, while for oils of bitter orange, sandalwood and camphor a stimulating effect on a growth of fungus B. cinerea was determined.

  3. Dichrostachys cinerea as a possible energy crop - facts and figures

    Energy Technology Data Exchange (ETDEWEB)

    Pedroso, Daniel Travieso [University of Camaguey, Camaguey (Cuba); Kaltschmitt, Martin [Hamburg University of Technology, Hamburg (Germany)

    2012-03-15

    Biomass contributes already with more than 10% to cover the global energy demand. This contribution will continue to grow in the years to come due to increasing fossil fuel prices and climate protection. To make this happen, additional sustainable biomass resources must become available to be used as a source of energy. Against this background, the goal of this paper is it to analyse the properties of Dichrostachys cinerea (Marabu) as an energy crop. The investigation shows that this wood is characterised by properties comparable with other types of woody biomass with a longer crop period. Only the ash content is slightly higher. In addition, the airborne emissions released during combustion are relatively low in general. Thus, wood from D. cinerea (Marabu) can be seen as a promising fuel. (orig.)

  4. Production, purification, and characterization of a major Penicillium glabrum xylanase using Brewer's spent grain as substrate.

    Science.gov (United States)

    Knob, Adriana; Beitel, Susan Michelz; Fortkamp, Diana; Terrasan, César Rafael Fanchini; de Almeida, Alex Fernando

    2013-01-01

    In recent decades, xylanases have been used in many processing industries. This study describes the xylanase production by Penicillium glabrum using brewer's spent grain as substrate. Additionally, this is the first work that reports the purification and characterization of a xylanase using this agroindustrial waste. Optimal production was obtained when P. glabrum was grown in liquid medium in pH 5.5, at 25 °C, under stationary condition for six days. The xylanase from P. glabrum was purified to homogeneity by a rapid and inexpensive procedure, using ammonium sulfate fractionation and molecular exclusion chromatography. SDS-PAGE analysis revealed one band with estimated molecular mass of 18.36 kDa. The optimum activity was observed at 60 °C, in pH 3.0. The enzyme was very stable at 50 °C, and high pH stability was verified from pH 2.5 to 5.0. The ion Mn(2+) and the reducing agents β -mercaptoethanol and DTT enhanced xylanase activity, while the ions Hg(2+), Zn(2+), and Cu(2+) as well as the detergent SDS were strong inhibitors of the enzyme. The use of brewer's spent grain as substrate for xylanase production cannot only add value and decrease the amount of this waste but also reduce the xylanase production cost.

  5. Production, Purification, and Characterization of a Major Penicillium glabrum Xylanase Using Brewer's Spent Grain as Substrate

    Directory of Open Access Journals (Sweden)

    Adriana Knob

    2013-01-01

    Full Text Available In recent decades, xylanases have been used in many processing industries. This study describes the xylanase production by Penicillium glabrum using brewer's spent grain as substrate. Additionally, this is the first work that reports the purification and characterization of a xylanase using this agroindustrial waste. Optimal production was obtained when P. glabrum was grown in liquid medium in pH 5.5, at 25 °C, under stationary condition for six days. The xylanase from P. glabrum was purified to homogeneity by a rapid and inexpensive procedure, using ammonium sulfate fractionation and molecular exclusion chromatography. SDS-PAGE analysis revealed one band with estimated molecular mass of 18.36 kDa. The optimum activity was observed at 60 °C, in pH 3.0. The enzyme was very stable at 50 °C, and high pH stability was verified from pH 2.5 to 5.0. The ion Mn2+ and the reducing agents β-mercaptoethanol and DTT enhanced xylanase activity, while the ions Hg2+, Zn2+, and Cu2+ as well as the detergent SDS were strong inhibitors of the enzyme. The use of brewer's spent grain as substrate for xylanase production cannot only add value and decrease the amount of this waste but also reduce the xylanase production cost.

  6. Novel thermostable endo-xylanase cloned and expressed from bacterium Geobacillus sp. WSUCF1.

    Science.gov (United States)

    Bhalla, Aditya; Bischoff, Kenneth M; Uppugundla, Nirmal; Balan, Venkatesh; Sani, Rajesh K

    2014-08-01

    A gene encoding a GH10 endo-xylanase from Geobacillus sp. WSUCF1 was cloned and expressed in Escherichia coli. Recombinant endo-xylanase (37kDa) exhibited high specific activity of 461.0U/mg of protein. Endo-xylanase was optimally active on birchwood xylan at 70°C and pH 6.5. The endo-xylanase was found to be highly thermostable at 50 and 60°C, retaining 82% and 50% of its original activity, respectively, after 60h. High xylan conversions (92%) were obtained with oat-spelt xylan hydrolysis. Higher glucan and xylan conversions were obtained on AFEX-treated corn stover with an enzyme cocktail containing WSUCF1 endo-xylanase (71% and 47%) as compared to enzyme cocktail containing commercial fungal endo-xylanase (64% and 41%). High specific activity, active at high pH's, wide substrate specificity, and higher hydrolytic activity on recalcitrant lignocellulose, make this endo-xylanase a suitable candidate for biofuel and bioprocess industries.

  7. Molecular characterization of a Xylanase-producing fungus isolated from fouled soil.

    Science.gov (United States)

    Sakthiselvan, Punniavan; Naveena, Balakrishnan; Partha, Nagarajan

    2014-01-01

    Xylanase (EC 3. 2. 1. 8), hydrolyzes xylo-oligosaccharides into D-xylose and required for complete hydrolysis of native cellulose and biomass conversion. It has broad range of applications in the pulp and paper, pharmaceutical and Agri-food industries. Fifty fungal species were isolated from the fouled soil around an oil refinery and screened for the production of xylanase enzyme by enrichment culture techniques. The isolated fungal strain was identified as Hypocrea lixii SS1 based on the results of biochemical tests and 18s rRNA sequencing. The phylogenetic tree was constructed using the MEGA 5 software. Further, Hypocrea lixii SS1 was tested for the ability to utilize the sunflower oil sludge (waste from the oil industry) as the sole carbon source for xylanase production. The growth characteristics of Hypocrea lixii SS1 were also studied and maximum growth was found on the 7(th) day of incubation. The fungus showed a remarkable xylanase production of 38.9 U/mL. Xylanase was purified using a combination of 0-50% NH₄SO₂ precipitation, DEAE-sepharose and Sephacryl S-200 chromatography. Single peak obtained in RP-HPLC confirms the purity of xylanase. Further the enzyme produced was affirmed as xylanase with its molecular weight (29 kDa) using SDS-PAGE.

  8. Production and Partial Characterization of an Alkaline Xylanase from a Novel Fungus Cladosporium oxysporum

    Directory of Open Access Journals (Sweden)

    Guo-Qiang Guan

    2016-01-01

    Full Text Available A new fungus Cladosporium oxysporum GQ-3 producing extracellular xylanase was isolated from decaying agricultural waste and identified based on the morphology and comparison of internal transcribed spacer (ITS rDNA gene sequence. C. oxysporum produced maximum xylanase activity of 55.92 U/mL with wheat bran as a substrate and NH4Cl as a nitrogen source. Mg2+ improved C. oxysporum xylanase production. Partially purified xylanase exhibited maximum activity at 50°C and pH 8.0, respectively, and showed the stable activity after 2-h treatment in pH 7.0–8.5 or below 55°C. Mg2+ enhanced the xylanase activity by 2% while Cu2+ had the highest inhibition ratio of 57.9%. Furthermore, C. oxysporum xylanase was resistant to most of tested neutral and alkaline proteases. Our findings indicated that Cladosporium oxysporum GQ-3 was a novel xylanase producer, which could be used in the textile processes or paper/feed industries.

  9. Thermo-mechanical and micro-structural properties of xylanase containing whole wheat bread

    Directory of Open Access Journals (Sweden)

    G. Ghoshal

    2016-12-01

    Full Text Available Xylanase is a hemicellulase that can hydrolyses the complex polysaccharides. Hemicelluloses are main components of cell walls of cereal grains. Moreover, hemicelluloses are considered as potential sources of mono- and oligosaccharides. In this study, influence of xylanase on the physicochemical properties and sensory qualities of the whole wheat bread during storage was investigated. Studies of whole wheat bread on microstructure, texture, thermotics, Scanning Electron Microscopic (SEM, X-Ray Diffraction (XRD were conducted at ambient temperature of 25 and 4 °C respectively. During storage at different temperatures, bread containing xylanase exhibited less firmness but larger volume with whiter crumb color and longer shelf life as compared to control bread. Results of firmness, enthalpy, Fourier Transformation Infra Red (FTIR and X-Ray Diffraction (XRD studies suggested a lower staling rate of bread containing xylanase as compared to control one. Bread containing xylanase showed a smoother surface and more uniform pore size than the control. Significant differences in microstructure of control and bread containing xylanase were observed which might be attributed due to the change in water starch gluten interaction. These differences were also found to be interrelated to the textural properties of bread. Better sensory features were achieved in bread containing xylanase.

  10. Molecular characterization of a Xylanase-producing fungus isolated from fouled soil

    Directory of Open Access Journals (Sweden)

    Punniavan Sakthiselvan

    2014-12-01

    Full Text Available Xylanase (EC 3. 2. 1. 8, hydrolyzes xylo-oligosaccharides into D-xylose and required for complete hydrolysis of native cellulose and biomass conversion. It has broad range of applications in the pulp and paper, pharmaceutical and Agri-food industries. Fifty fungal species were isolated from the fouled soil around an oil refinery and screened for the production of xylanase enzyme by enrichment culture techniques. The isolated fungal strain was identified as Hypocrea lixii SS1 based on the results of biochemical tests and 18s rRNA sequencing. The phylogenetic tree was constructed using the MEGA 5 software. Further, Hypocrea lixii SS1 was tested for the ability to utilize the sunflower oil sludge (waste from the oil industry as the sole carbon source for xylanase production. The growth characteristics of Hypocrea lixii SS1 were also studied and maximum growth was found on the 7th day of incubation. The fungus showed a remarkable xylanase production of 38.9 U/mL. Xylanase was purified using a combination of 0-50% NH4SO2 precipitation, DEAE-sepharose and Sephacryl S-200 chromatography. Single peak obtained in RP-HPLC confirms the purity of xylanase. Further the enzyme produced was affirmed as xylanase with its molecular weight (29 kDa using SDS-PAGE.

  11. Botrytis cinerea: Causal agent of small fruit grey mould

    OpenAIRE

    Tanović, Brankica

    2012-01-01

    Small fruits growing in Serbia is an important and profitable business. However, disease causal agents, pests and weeds often threaten production profitability. A common problem in production of most important small fruit species is a polyfagous, phytopathogenic fungal species Botrytis cinerea, the causal agent of grey mould disease of fruits. Present knowledge on the causal agent, its morphological, ecological and epidemiological characteristics are systematized in the paper. Infection proce...

  12. The microbial oxidation of (-)-beta-pinene by Botrytis cinerea.

    Science.gov (United States)

    Farooq, Afgan; Choudhary, M Iqbal; Tahara, Satoshi; Rahman, Atta-ur; Başer, K Hüsnü Can; Demirci, Fatih

    2002-01-01

    (-)-beta-pinene, a flavor and fragrance monoterpene is an important constituent of essential oils of many aromatic plants. It was oxidized by a plant-pathogenic fungus, Botrytis cinerea to afford four metabolites characterized as (-)-6a-hydroxy-beta-pinene, (-)-4beta,5beta-dihydroxy-beta-pinene, (-)-2beta,3beta-dihydroxypinane, and (-)-4beta-hydroxy-beta-pinene-6-one by detailed spectroscopic studies along with other known metabolites.

  13. Biotransformation of (-)-a-pinene by Botrytis cinerea.

    Science.gov (United States)

    Farooq, Afgan; Tahara, Satoshi; Choudhary, M Iqbal; Atta-ur-Rahman; Ahmed, Zafar; Hüsnü, Can Başer K; Demirci, Fatih

    2002-01-01

    (-)-alpha-Pinene (1), a major constituent of many aromatic plants was biotransformed by the plant pathogenic fungus, Botrytis cinerea to afford three new metabolites, characterized as 3beta-hydroxy-(-)-beta-pinene (10%) (3), 9-hydroxy-(-)-a-pinene (12%) (4), 4beta-hydroxy-(-)-alpha-pinene-6-one (16%) (5) by physical and spectroscopic methods. A known metabolite verbenone (2) was also obtained.

  14. Characterization and biotechnological application of recombinant xylanases from Aspergillus nidulans.

    Science.gov (United States)

    Maitan-Alfenas, Gabriela P; Oliveira, Mariana B; Nagem, Ronaldo A P; de Vries, Ronald P; Guimarães, Valéria M

    2016-10-01

    Two xylanases from Aspergillus nidulans, XlnB and XlnC, were expressed in Pichia pastoris, purified and characterized. XlnB and XlnC achieved maximal activities at 60°C and pH 7.5 and at 50°C and pH 6.0, respectively. XlnB showed to be very thermostable by maintaining 50% of its original activity after 49h incubated at 50°C. XlnB had its highest activity against wheat arabinoxylan while XlnC had the best activity against beechwood xylan. Both enzymes were completely inhibited by SDS and HgCl2. Xylotriose at 1mg/ml also totally inibited XlnB activity. TLC analysis showed that the main product of beechwood xylan hydrolysis by XlnB and XlnC was xylotetraose. An additive effect was shown between XlnB and XlnC and the xylanases of two tested commercial cocktails. Sugarcane bagasse saccharification results showed that these two commercial enzymatic cocktails were able to release more glucose and xylose after supplementation with XlnB and XlnC.

  15. Highly thermostable xylanase production from a thermophilic Geobacillus sp. strain WSUCF1 utilizing lignocellulosic biomass

    Directory of Open Access Journals (Sweden)

    Aditya eBhalla

    2015-06-01

    Full Text Available AbstractEfficient enzymatic hydrolysis of lignocellulose to fermentable sugars requires a complete repertoire of biomass deconstruction enzymes. Hemicellulases play an important role in hydrolyzing hemicellulose component of lignocellulose to xylo-oligosaccharides and xylose. Thermostable xylanases have been a focus of attention as industrially important enzymes due to their long shelf life at high temperatures. Geobacillus sp. strain WSUCF1 produced thermostable xylanase activity (crude xylanase cocktail when grown on xylan or various inexpensive untreated and pretreated lignocellulosic biomasses such as prairie cord grass and corn stover. The optimum pH and temperature for the crude xylanase cocktail were 6.5 and 70ºC, respectively. The WSUCF1 crude xylanase was found to be highly thermostable with half-lives of 18 and 12 days at 60 and 70ºC, respectively. At 70ºC, rates of xylan hydrolysis were also found to be better with the WSUCF1 secretome than those with commercial enzymes, i.e., for WSUCF1 crude xylanase, CellicHTec2, and AccelleraseXY, the percent xylan conversions were 68.9, 49.4, and 28.92, respectively. To the best of our knowledge, WSUCF1 crude xylanase cocktail is among the most thermostable xylanases produced by thermophilic Geobacillus spp. and other thermophilic microbes (optimum growth temperature ≤70ºC. High thermostability, activity over wide range of temperatures, and better xylan hydrolysis than commercial enzymes make WSUCF1 crude xylanase suitable for thermophilic lignocellulose bioconversion processes.

  16. Fungicide resistance phenotypes in Botrytis cinerea populations from blueberries in California and Washington

    Science.gov (United States)

    Gray mold caused by Botrytis cinerea is a major postharvest disease of blueberries grown in the Central Valley of California (CA) and western Washington State (WA). Understanding fungicide- resistant phenotypes of B. cinerea is important to the development of preharvest fungicide programs for contro...

  17. Biologische Bekämpfung von Botrytis cinerea mit Ulocladium atrum in Reben und Cyclamen

    NARCIS (Netherlands)

    Schoene, P.; Köhl, J.

    1999-01-01

    Botrytis cinerea causes world-wide yield loss in many crops and fungicide resistant strains impede the control. Beside chemical control, antagonistic bacteria and fungi were used for biological control. The saprophytic hyphomycet Ulocladium atrum has shown an antagonistic effect against B. cinerea a

  18. Meningitis and Bacteremia Due to Neisseria cinerea following a Percutaneous Rhizotomy of the Trigeminal Ganglion.

    Science.gov (United States)

    von Kietzell, M; Richter, H; Bruderer, T; Goldenberger, D; Emonet, S; Strahm, C

    2016-01-01

    Neisseria cinerea is a human commensal. The first known case of meningitis and bacteremia due to Neisseria cinerea following percutaneous glycerol instillation of the trigeminal ganglion is reported. Conventional phenotypic methods and complete 16S RNA gene sequencing accurately identified the pathogen. Difficulties in differentiation from pathogenic neisseriae are discussed.

  19. Fatal bacteremia by neisseria cinerea in a woman with myelodysplastic syndrome: a case report.

    Science.gov (United States)

    Zhu, Xiaofei; Li, Min; Cao, Huiling; Yang, Xuewen

    2015-01-01

    Neisseria cinerea has been rarely found in blood cultures. In this study, we are reporting a case of a Myelodysplastic Syndrome (MDS) patient in whose blood Neisseria cinerea was found and led a fatal consequence. This case will call our attentions to the uncommon pathogens in the pathogenicity of end-stage patients.

  20. Meningitis and Bacteremia Due to Neisseria cinerea following a Percutaneous Rhizotomy of the Trigeminal Ganglion

    OpenAIRE

    von Kietzell, M.; Richter, H.; Bruderer, T.; Goldenberger, D.; Emonet, S; Strahm, C.

    2015-01-01

    Neisseria cinerea is a human commensal. The first known case of meningitis and bacteremia due to Neisseria cinerea following percutaneous glycerol instillation of the trigeminal ganglion is reported. Conventional phenotypic methods and complete 16S RNA gene sequencing accurately identified the pathogen. Difficulties in differentiation from pathogenic neisseriae are discussed.

  1. Fatal bacteremia by neisseria cinerea in a woman with myelodysplastic syndrome: a case report

    OpenAIRE

    Zhu, Xiaofei; Li, Min; Cao, Huiling; Yang, Xuewen

    2015-01-01

    Neisseria cinerea has been rarely found in blood cultures. In this study, we are reporting a case of a Myelodysplastic Syndrome (MDS) patient in whose blood Neisseria cinerea was found and led a fatal consequence. This case will call our attentions to the uncommon pathogens in the pathogenicity of end-stage patients.

  2. Purification and characterization of a moderately thermostable xylanase from Bacillus sp. strain SPS-0.

    Science.gov (United States)

    Bataillon; Nunes Cardinali A; Castillon; Duchiron

    2000-02-01

    A Bacillus spp. strain SPS-0, isolated from a hot spring in Portugal, produced an extracellular xylanase upon growth on wheat bran arabinoxylan. The enzyme was purified to homogeneity by ammonium sulfate precipitation, anion exchange, gel filtration, and affinity chromatography. The optimum temperature and pH for activity was 75 degrees C and 6.0. Xylanase was stable up to 70 degrees C for 4 h at pH 6.0 in the presence of xylane. Xylanase was completely inhibited by the Hg(2+) ions. beta-Mercaptoethanol, dithiothreitol, and Mn(2+) stimulated the xylanase activity. The products of birchwood xylan hydrolysis were xylose, xylobiose, xylotriose, and xylotetraose. Kinetic experiments at 60 degrees C and pH 6.0 gave V(max) and K(m)values of 2420 nkat/mg and 0.7 mg/ml.

  3. [Cellulase and xylanase activities of Fusarium Lk:Fr. genus fungi of different trophic groups].

    Science.gov (United States)

    Kurchenko, I M; Sokolova, O V; Zhdanova, N M; Iarynchyn, A M; Iovenko, O M

    2008-01-01

    A comparative analysis of cellulase and xylanase activities of 26 fungal strains of phytopathogenic, saprophytic and endophytic Fusarium species has been realized using the qualitative reactions. The rare of their linear growth on the media with carboxymethyl cellulose or xylane has been studied. It was shown that the fungi of genus Fusarium belonging to different trophic groups possessed low activities of investigated enzymes as a whole, but in endophytic strains their levels were lower than in phytopathogenic ones. At the same time the distinct strain dependence of cellulase and xylanase activities was fixed in the fungi of different trophic groups. As far as the cellulase and xylanase activities in phytopathogenic isolates varied from complete absence to high levels, and since the activity maximum for each of the investigated strains was observed in different growth terms the conclusion was made that the cellulase and xylanase activities could not be considered as possible markers of the fungal isolate pathogenicity on the strain level.

  4. Influence of agitation speeds and aeration rates on the Xylanase activity of Aspergillus niger SS7

    Directory of Open Access Journals (Sweden)

    Yasser Bakri

    2011-08-01

    Full Text Available In this study, the effect of agitation and aeration rates on xylanase activity of Aspergillus niger SS7 in 3-litre stirred tank bioreactor was investigated. The agitation rates tested were 100, 200 and 300 rpm at each airflow rates of 0.5, 1.0 and 1.5 vvm. The maximum xylanase activity in mono- agitator system was at the agitation speed of 200 rpm and aeration rate of 1.0 vvm. In bi-agitator system, at low agitation speed (100 rpm, the xylanase activity was enhanced by 13% compared to mono- agitator system for an aeration rate of 1.0 vvm. Xylanase productivity in continuous culture was higher by approximately 3.5 times than in batch culture.

  5. Mutation-Screening in Xylanase-Producing Strains by Ion Implantation

    Institute of Scientific and Technical Information of China (English)

    李市场; 吴敏; 姚建铭; 潘仁瑞; 余增亮

    2005-01-01

    With ion implantation (N+, energy 10 keV and dosage 1.56×1015N+ cm-2), a high xylanase-producing strain Aspergillus niger N212 was selected. Based on an orthogonal experiment, an optimal fermentation condition was designed for this high-yield strain. The suitable medium was composed of 8% corncob; 1.0% wheat bran; 0.1% TWEEN20; 0.5% (NH4)2SO4;3.0×10-4. At present, under our experiment condition, xylanase activity of Aspergillus niger N212 reached a level of 600 IU/ml, almost increased by 100% in xylanase production and the time of yielding xylanase was largely reduced to 12 h at 28 ℃.

  6. Screening and production study of microbial xylanase producers from Brazilian Cerrado.

    Science.gov (United States)

    Alves-Prado, Heloiza Ferreira; Pavezzi, Fabiana Carina; Leite, Rodrigo Simões Ribeiro; de Oliveira, Valéria Maia; Sette, Lara Durães; Dasilva, Roberto

    2010-05-01

    Hemicelluloses are polysaccharides of low molecular weight containing 100 to 200 glycosidic residues. In plants, the xylans or the hemicelluloses are situated between the lignin and the collection of cellulose fibers underneath. The xylan is the most common hemicellulosic polysaccharide in cell walls of land plants, comprising a backbone of xylose residues linked by beta-1,4-glycosidic bonds. So, xylanolytic enzymes from microorganism have attracted a great deal of attention in the last decade, particularly because of their biotechnological characteristics in various industrial processes, related to food, feed, ethanol, pulp, and paper industries. A microbial screening of xylanase producer was carried out in Brazilian Cerrado area in Selviria city, Mato Grosso do Sul State, Brazil. About 50 bacterial strains and 15 fungal strains were isolated from soil sample at 35 degrees C. Between these isolated microorganisms, a bacterium Lysinibacillus sp. and a fungus Neosartorya spinosa as good xylanase producers were identified. Based on identification processes, Lysinibacillus sp. is a new species and the xylanase production by this bacterial genus was not reported yet. Similarly, it has not reported about xylanase production from N. spinosa. The bacterial strain P5B1 identified as Lysinibacillus sp. was cultivated on submerged fermentation using as substrate xylan, wheat bran, corn straw, corncob, and sugar cane bagasse. Corn straw and wheat bran show a good xylanase activity after 72 h of fermentation. A fungus identified as N. spinosa (strain P2D16) was cultivated on solid-state fermentation using as substrate source wheat bran, wheat bran plus sawdust, corn straw, corncob, cassava bran, and sugar cane bagasse. Wheat bran and corncobs show the better xylanase production after 72 h of fermentation. Both crude xylanases were characterized and a bacterial xylanase shows optimum pH for enzyme activity at 6.0, whereas a fungal xylanase has optimum pH at 5.0-5.5. They were

  7. Heat-induced oxidative injury contributes to inhibition of Botrytis cinerea spore germination and growth.

    Science.gov (United States)

    Zhao, Wei; Wisniewski, Michael; Wang, Wenjie; Liu, Jia; Liu, Yongsheng

    2014-03-01

    The inhibitory effect of heat treatment (HT) on Botrytis cinerea, a major postharvest fungal pathogen, and the possible mode of action were investigated. Spore germination and germ tube elongation of B. cinerea were both increasingly and significantly inhibited by HT (43 °C) for 10, 20 or 30 min. HT-induced gene expression of NADPH oxidase A, resulted in the intracellular accumulation of reactive oxygen species. HT-treated B. cinerea spores exhibited higher levels of oxidative damage to proteins and lipids, compared to the non-HT control. These findings indicate that HT resulted in oxidative damage which then played an important role in the inhibitory effect on B. cinerea. In the current study, HT was effective in controlling gray mold, caused by B. cinerea, in pear fruits. Understanding the mode of action by which HT inhibits fungal pathogens will help in the application of HT for management of postharvest fungal diseases of fruits and vegetables.

  8. Prevalence and persistence of Neisseria cinerea and other Neisseria spp. in adults.

    Science.gov (United States)

    Knapp, J S; Hook, E W

    1988-05-01

    Neisseria cinerea is a commensal Neisseria sp. which was first described in 1906 but was subsequently misclassified as a subtype of Branhamella catarrhalis. N. cinerea resembles Neisseria gonorrhoeae in both cultural and biochemical characteristics and, thus, may also have been misidentified as N. gonorrhoeae. Of 202 patients whose oropharynges were colonized by Neisseria spp., N. cinerea was isolated in 57 (28.2%) patients, including 25 (30.1%) of 83 women, 22 (23.9%) of 92 heterosexual men, and 10 (37.0%) of 27 homosexual men in Seattle, Wash., in 1983 to 1984. N. cinerea was isolated from the urethra of only one (1.1%) patient. The oropharynges of many individuals were colonized persistently by strains of N. cinerea and other Neisseria spp.

  9. Mutagenesis and Screening of High Yield Xylanase Production Strain of Aspergillus usamii by Microwave Irradiation

    Institute of Scientific and Technical Information of China (English)

    李永泉; 陈时飞; 岑沛霖

    2003-01-01

    A high yield xylanase producing strain, A. usamii L336-23, was screened out from its parent strain A.usamii L336 after microwave irradiation. Its productivity of xylanase activity increased by 35.7% from 21000μ·m1-1 to 28500μ·m1-1 and was stable after frequent subcultures and storage for more than two months.The mechanism of microwave mutation was also discussed.

  10. A Highly Thermostable Xylanase from Stenotrophomonas maltophilia: Purification and Partial Characterization

    Directory of Open Access Journals (Sweden)

    Abhay Raj

    2013-01-01

    Full Text Available Seven xylanolytic bacterial strains were isolated from saw-dust dump soil. The bacterial strain X6 was selected on the basis of the highest xylanase activity with no cellulase contamination. It was identified as Stenotrophomonas maltophilia by biochemical tests and 16S rRNA gene sequencing approach. Xylanase production studies by S. maltophilia on different commercial xylans and agro-industrial residues suggested that wheat bran was the best carbon source for xylanase production (26.4 ± 0.6 IU/mL. The studies with inorganic and organic nitrogen sources suggested yeast extract as the best support for xylanase production (25 ± 0.6 IU/mL. Maximum xylanase production was observed at initial medium pH = 8.0 (23.8 ± 0.4 IU/mL with production at pH = 7.0 and pH = 9.0 being almost comparable. Xylanase produced by S. maltophilia was purified to homogeneity using ammonium sulfate precipitation, gel filtration, and ion exchange chromatography. The final purification was 5.43-fold with recovery of 19.18%. The molecular weight of the purified xylanase protein was ~142 kDa. Both crude and purified xylanase had good stability at pH = 9.0 and 80°C with activity retention greater than 90% after 30 min incubation. The enzyme stability at high temperature and alkaline pH make it potentially effective for industrial applications.

  11. Bacterial xylanase expression in mammalian cells and transgenic mice.

    Science.gov (United States)

    Fontes, C M; Ali, S; Gilbert, H J; Hazlewood, G P; Hirst, B H; Hall, J

    1999-06-11

    The energy which simple-stomached livestock can derive from dietary plant material is limited by the lack of plant polysaccharide degrading enzymes in their gastro-intestinal (GI) tract and the inefficient microbial fermentation of such material in their hind-gut. In poultry the non-starch polysaccharides found in cereal grains can also impair normal digestive function as they form viscous gels in the GI tract inhibiting the breakdown and absorption of nutrients. The nutrition of such livestock could, therefore, be improved by the introduction of enzymes able to degrade plant polysaccharides in the small intestine. We describe the expression of a xylanase, XYLY', from the bacterium Clostridium thermocellum in mammalian cells and the exocrine pancreas of transgenic mice. The enzyme is synthesised, secreted and functionally active in the eukaryote system. This work demonstrates the feasibility of generating animals with the endogenous capacity to depolymerise the xylan component of hemi-cellulose.

  12. Molecular breeding of transgenic rice expressing a xylanase domain of the xynA gene from Clostridium thermocellum.

    Science.gov (United States)

    Kimura, T; Mizutani, T; Tanaka, T; Koyama, T; Sakka, K; Ohmiya, K

    2003-09-01

    The gene encoding the catalytic domain of thermostable xylanase from Clostridium thermocellum F1 was expressed in rice plants under the control of a constitutive promoter. The gene encoding Xylanase A was modified to encode the catalytic domain of family 11 xylanase without the signal sequence (xynA1), and was introduced into rice plants and expressed under the control of a modified cauliflower mosaic virus 35S promoter. Zymogram analysis indicated that the recombinant xylanase was produced in rice plants. The xynA1 gene was stably expressed in rice straw and seed grains. No phenotypic effect of xylanase expression was noted. The enzyme was detected in the desiccated grain. High levels of enzyme activity were maintained in the cell-free extract during incubation at 60 degrees C for 24 h. The results indicated that high levels of xylanase can be produced in rice plants.

  13. Removal of hexenuronic acid by xylanase to reduce adsorbable organic halides formation in chlorine dioxide bleaching of bagasse pulp.

    Science.gov (United States)

    Nie, Shuangxi; Wang, Shuangfei; Qin, Chengrong; Yao, Shuangquan; Ebonka, Johnbull Friday; Song, Xueping; Li, Kecheng

    2015-11-01

    Xylanase-aided chlorine dioxide bleaching of bagasse pulp was investigated. The pulp was pretreated with xylanase and followed a chlorine dioxide bleaching stage. The ATR-FTIR and XPS were employed to determine the surface chemistry of the control pulp, xylanase treated and chlorine dioxide treated pulps. The hexenuronic acid (HexA) could obviously be reduced after xylanase pretreatment, and the adsorbable organic halides (AOX) were reduced after chlorine dioxide bleaching. Compared to the control pulp, AOX could be reduced by 21.4-26.6% with xylanase treatment. Chlorine dioxide demand could be reduced by 12.5-22% to achieve the same brightness. The ATR-FTIR and XPS results showed that lignin and hemicellulose (mainly HexA) were the main source for AOX formation. Xylanase pretreatment could remove HexA and expose more lignin, which decreased the chlorine dioxide demand and thus reduced formation of AOX.

  14. [Xylanase and cellulase of fungus Cerrena unicolor VKM F-3196: production, properties, and applications for the saccharification of plant material].

    Science.gov (United States)

    Belova, O V; Lisov, A V; Vinokurova, N G; Kostenevich, A A; Sapunova, L I; Lobanok, A G; Leont'evskiĭ, A A

    2014-01-01

    Under the conditions of submerged cultivation in a medium containing microcrystalline cellulose, the Cerrena unicolor VKM F-3196 basidiomycete is capable of producing xylanase and cellulase. Electrophoretically homogeneous cellulase and xylanase were obtained using ion exchange and hydrophobic chromatography. The molecular weight of both cellulase and xylanase was -44 kDa. It was shown that xylanase catalyzed the hydrolysis of xylan with the production of xylose, xylobiose, and xylotetrose and it exhibited properties of endoxylanases. Cellulase hydrolyzed carboxymethylcellulose, xylan, and microcrystalline cellulose with the formation of cellotriose and cellotetraose. For both enzymes, the pH optimum was -4.0. The enzymes exhibited moderate thermostability: xylanase retained 35% of the initial activity for an hour at 60 degrees C; cellulase, 10% under the same conditions. Xylanase, cellulose, and a mixture of these enzymes saccharified plant material (wheat, rye, wheat middling, and oat), indicating the possible use of these enzymes in biotechnology.

  15. Xylanase production by a local fungal isolate, Aspergillus niger USM AI 1 via solid state

    Directory of Open Access Journals (Sweden)

    Ibrahim Che Omar

    2005-03-01

    Full Text Available Isolate USM A1 I which was identified to be Aspergillus niger was selected as a potential producer of xylanase via a solid state fermentation system (SSF using palm kernel cake (PKC as substrate. The modification of the physical conditions of the SSF system indicated that the xylanase activity was 23.97 U/g PKC at the moisture ratio of 1:0.75 of PKC: moistening agent with the inoculum size of 1¥104 spores/ml and cultivated at the ambient temperature (28±3ºC. The supplementation of additional carbon and nitrogen sources in the PKC medium could enhance enzyme productivity. The maximum production of xylanase and growth obtained with the supplementation of xylose at 0.75% (w/w were 25.40 U/g and 1.69 mg glucosamine/ g PKC. Moreover, the presence of NaNO3 at 0.075% (w/w as additional nitrogen source further enhanced xylanase production to 33.99 U/g PKC although the growth remained unchanged at about 1.67 mg glucosa- mine/g PKC. The optimized conditions showed an increased xylanase production by 157% compared to before the optimization of the SSF system. The xylanase productivity was 23.12 U/mg glucosamine after optimization and 11.72 U/mg glucosamine before optimization.

  16. Attachment capability of antagonistic yeast Rhodotorula glutinis to Botrytis cinerea contributes to biocontrol efficacy

    Directory of Open Access Journals (Sweden)

    Boqiang eLi

    2016-05-01

    Full Text Available Rhodotorula glutinis as an antagonism show good biocontrol performance against various postharvest diseases in fruits. In the present study, strong attachment capability of R. glutinis to spores and hyphae of Botrytis cinerea was observed. Further analysis showed that certain protein components on the yeast cell surface played critical role during the interaction between R. glutinis and B. cinerea. The components mainly distributed at the poles of yeast cells and might contain glycosylation modification, as tunicamycin treated yeast cells lost attachment capability to B. cinerea. To investigate contributions of attachment capability of R. glutinis to its biocontrol efficacy, yeast cells were mutagenized with 3% methane-sulfonic acid ethyl ester (EMS, and a mutant CE4 with stable non-attaching phenotype was obtained. No significant difference was found on colony, cell morphology, reproductive ability and capsule formation between the mutant and wild type. However, there was a distinct difference in India ink positive staining patterns between the two strains. Moreover, wild type strain of R. glutinis showed better performance on inhibiting spore germination and mycelial growth of B. cinerea than CE4 strain when yeast cells and B. cinerea were co-cultured in vitro. In biocontrol assay, both wild type and CE4 strains showed significant biocontrol efficacy against gray mould caused by B. cinerea in apple fruit, whereas, control effect of CE4 strain was lower than that of wild type. Our findings provided new evidences that attachment capability of R. glutinis to B. cinerea contributed to its biocontrol efficacy.

  17. Functional characterization of Penicillium occitanis Pol6 and Penicillium funiculosum GH11 xylanases.

    Science.gov (United States)

    Driss, Dorra; Berrin, Jean Guy; Juge, Nathalie; Bhiri, Fatma; Ghorbel, Raoudha; Chaabouni, Semia Ellouz

    2013-08-01

    Xylanases are hemicellulolytic enzymes, which are responsible for the degradation of heteroxylans constituting the lignocellulosic plant cell wall. Xylanases from the GH11 family are considered as true xylanases because of their high substrate specificity. In order to study in depth a crucial difference in the thumb region between two closely related xylanases from Penicillium in terms of kinetic parameters and inhibition sensitivity, the GH11 xylanases from Penicillium occitanis Pol6 (PoXyn3) and from Penicillium funiculosum (PfXynC) were heterologously expressed in Pichia pastoris. The PoXyn3 and PfXynC cDNAs encoding mature xylanases were cloned into pGAPZαA vectors and integrated into the genome of P. pastoris X-33 under the control of the glyceraldehyde 3-phosphate dehydrogenase constitutive promoter. PfXynC was expressed as a His-tagged recombinant protein and purified from the supernatant homogeneity by a one-step purification protocol using immobilized metal affinity chromatography. The recombinant PoXyn3 was purified using a single anion-exchange chromatography. The purified recombinant enzymes were optimally active at 45°C and pH 4.0 for PoXyn3 and 40°C and pH 3.0 for PfXynC. The measured kinetic parameters (k(cat) and Vmax) showed that PfXynC was five times more active than PoXyn3 irrespective of the substrate whereas the apparent affinity (K(m)) was similar. The recombinant enzymes showed distinct sensitivity to the Triticum aestivum xylanase inhibitor TAXI-I.

  18. Ozone and infection of geranium flowers by Botrytis cinerea

    Energy Technology Data Exchange (ETDEWEB)

    Manning, W.J.; Feder, W.A.; Perkins, I.

    1970-01-01

    Flowering plants of geranium cultivars were exposed to 0.2, 0.35, and 0.55 ppm ozone for 4-hr periods at 20/sup 0/C in a greenhouse fumigation chamber. Three fully-opened flower heads were sprayed with a spore suspension of Botrytis cinerea at 2000, 1000, or 500 spores/ml immediately before exposure to ozone began. Sterile distilled water was sprayed on noninoculated flower heads. All flowers were examined for evidence of infection 24 hr after the end of the ozone-exposure periods. All flower heads were then removed and placed in wet, loosely tied plastic bags and incubated at 20/sup 0/C for 72 hr, with examination at 24-hr intervals for evidence of infection. Ozone at 0.2 ppm did not injure the plants or prevent or inhibit flower infection by B. cinerea at all inoculum levels. Natural infection also occurred on some noninoculated flowers. Ozone at 0.35 ppm did not injure the plants or prevent infection, but did inhibit pathogenesis at the 500-spore/ml inoculum level and on noninoculated flowers. Ozone at 0.55 ppm caused moderate injury on all plants. Ozone at this level did not prevent infection, but did restrict pathogenesis on all inoculated and noninoculated flowers.

  19. 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

  20. Biological control of botrytis cinerea growth on apples stored in modified atmospheres

    DEFF Research Database (Denmark)

    Dock, Lise Lotte; Nielsen, Per Væggemose; Floros, John D.

    1998-01-01

    was set according to a centralcomposite experimental design involving five levels of O2 (1 to 15%)and CO2 (0 to 15%). Control samples under ambient conditions were alsoincluded. Without the antagonist, measurements of mold colony diameterover time showed that O2 had no effect on the growth of B. cinerea...... by about 6days at low levels of CO2. However, at high CO2 levels, O2 had noeffect. The strongest antagonistic effect was observed under ambientconditions. Overall, results showed that high CO2 atmospheres can slowthe growth of B. cinerea and that Erwinia sp. was an effectiveantagonist against B. cinerea...

  1. Efficiency of different xylanase preparations in diets for pekin ducks.

    Science.gov (United States)

    Timmler, R; Rodehutscord, M

    2001-01-01

    Four experiments were conducted with a total of 2288 pekin ducks. Day-old ducklings were group-penned on straw bedding and were fed complete, pelleted diets ad libitum for up to 49 days depending on experiment. In each experiment, starter diets (until day 21) and grower diets (from day 22) were used adequate in ME content and nutrient content. The sum of wheat, rye, and triticale amounted to at least 57% (starter diet) and 63% (grower diet), respectively. The inclusion level of wheat, rye, and triticale was different between experiments, with a maximum rye inclusion of 45%. Five different enzyme preparations all having, 1,4-beta-xylanase as the main activity were considered in this study with either one (2 preparations) or three (3 preparations) levels of supplementation. The effect of enzyme supplementation on ileal digesta viscosity was studied at the end of two experiments comprising 4 enzyme preparations. A significant reduction in digesta viscosity was determined for all preparations. The viscosity of digesta was higher in birds that were fed 45% rye in their diet as compared to those fed a diet based on triticale and wheat, even with enzyme supplementation. Differences in digesta viscosity were not reflected in growth or feed conversion data. In one experiment, the body weight of ducks on day 21 was significantly improved by enzyme supplementation. This effect disappeared with progress in experiment. In another experiment, feed intake was significantly improved with enzyme supplementation. Apart from this, no statistically significant improvement in performance could be detected. On overall average, the final BW of ducks fed an enzyme was (as compared to the unsupplemented control = 100), 100, and the feed conversion ratio was 101. There is no indication from the growth and feed conversion data that an enzyme effect becomes more pronounced with increasing inclusion rate of soluble NSP by rye. It is concluded that supplementary xylanases are efficient in

  2. 产木聚糖酶菌株的分离及木聚糖酶的固定化%Isolation of Xylanase Producing Strain and Immobilization of Xylanase

    Institute of Scientific and Technical Information of China (English)

    王瑞君

    2012-01-01

    从草鱼肠道中分离筛选出6株产木聚糖酶的菌株,从中挑选产酶量最大的菌株测定酶液活力,并采用壳聚糖、海藻酸钠等固定游离酶.结果表明,与游离酶相比,壳聚糖固定酶的最适pH向碱性方向偏移,海藻酸钠固定酶的最适pH向酸性方向偏移.与游离酶相比,固定化酶有较高的最适温度和较强的热稳定性.%6 xylanase producing strains were screened from the intestinal tract of grass carp, among -which the strain with the highest xylanase yield was adopted to detect the enzyme activity of xylanase. Furthermore, the xylanase was immobilized by chitosan and sodium alginate. The results showed that compared with free enzyme, the optimal pH for chitosan immobilized xylanase was higher while for sodium alginate immobilized xylanase was lower. The optimal temperature for the immobilized xylanase was higher than that of free enzyme. The immobilized xylanase also showed broader temperature adaptability.

  3. Stable production of thermotolerant xylanase B of Clostridium stercorarium in transgenic tobacco and rice.

    Science.gov (United States)

    Kimura, Tetsuya; Mizutani, Tomomi; Sun, Jia-Lin; Kawazu, Tetsu; Karita, Shuichi; Sakka, Makiko; Kobayashi, Yasuo; Ohmiya, Kunio; Sakka, Kazuo

    2010-01-01

    The xylanase B gene encoding a thermostable family 10 xylanase of Clostridium stercorarium was expressed in plants under the control of a constitutive promoter. Two forms of the xylanase B gene, the xynB gene encoding the full length of the xylanase B gene including the bacterial signal sequence and the xynBM gene without the signal sequence region, were introduced into tobacco BY-2 cells and tobacco plants respectively under the control of the cauliflower mosaic virus 35S promoter. Transgenic BY-2 cells and tobacco plants showed xylanase activity and normal growth. The recombinant enzyme produced in transgenic BY-2 cells harboring the xynB gene was secreted into the culture supernatant, and the recombinant enzyme produced in transgenic BY-2 cells harboring the xynBM gene was localized in the cells. In contrast to tobacco plants, expression of the xynB gene under the control of the rice actin promoter in rice plants was toxic to host cells. However, the recombinant XynBM accumulated in leaf cells, and no phenotypic effect of expression of the xynBM gene was observed. Enzyme activity was maintained in cell-free extracts of transgenic rice leaves at 60 degrees C for 72 h, and the recombinant XynBM degraded hemicellulosic polymers in cell-free extracts of transgenic rice leaves.

  4. Structure-Specificity Relationships of an Intracellular Xylanase from Geobacillus stearothermophilus

    Energy Technology Data Exchange (ETDEWEB)

    Solomon,V.; Teplitsky, A.; Shulami, S.; Zolotnitsky, G.; Shoham, Y.; Shoham, G.

    2007-01-01

    Geobacillus stearothermophilus T-6 is a thermophilic Gram-positive bacterium that produces two selective family 10 xylanases which both take part in the complete degradation and utilization of the xylan polymer. The two xylanases exhibit significantly different substrate specificities. While the extracellular xylanase (XT6; MW 43.8 kDa) hydrolyzes the long and branched native xylan polymer, the intracellular xylanase (IXT6; MW 38.6 kDa) preferentially hydrolyzes only short xylo-oligosaccharides. In this study, the detailed three-dimensional structure of IXT6 is reported, as determined by X-ray crystallography. It was initially solved by molecular replacement and then refined at 1.45 {angstrom} resolution to a final R factor of 15.0% and an R{sub free} of 19.0%. As expected, the structure forms the classical ({alpha}/{beta}){sub 8} fold, in which the two catalytic residues (Glu134 and Glu241) are located on the inner surface of the central cavity. The structure of IXT6 was compared with the highly homologous extracellular xylanase XT6, revealing a number of structural differences between the active sites of the two enzymes. In particular, structural differences derived from the unique subdomain in the carboxy-terminal region of XT6, which is completely absent in IXT6. These structural modifications may account for the significant differences in the substrate specificities of these otherwise very similar enzymes.

  5. Analysis of inducers of xylanase and cellulase activities production by Ganoderma applanatum LPB MR-56.

    Science.gov (United States)

    Salmon, Denise Naomi Xavier; Spier, Michele Rigon; Soccol, Carlos Ricardo; Vandenberghe, Luciana Porto de Souza; Weingartner Montibeller, Valesca; Bier, Mário César Jucoski; Faraco, Vincenza

    2014-08-01

    This manuscript describes the analysis of the effect of cellulose, carboxymethylcellulose (CMC), xylan, and xylose as inducers of cellulase and xylanase activity production by Ganoderma applanatum MR-56 and the optimization of their production in liquid cultures by statistical methods. The Plackett-Burman screening design was applied to identify the most significant inducers of xylanase and cellulase activities production by G. applanatum MR-56. The most significant effect on xylanase and cellulase activities production was exercised by cellulose, even if xylose and CMC were also effective at some times. The combined effect of cellulose, yeast extract, and pH was analyzed by a 2(3) factorial experimental design with four central points that showed that the maximum tested cellulose (1 % w/v) and yeast extract (5 g L(-1)) concentrations gave the maximum production of xylanase (8.24 U mL(-1)) and cellulase (3.29 U mL(-1)) activity at pH 6 and 4, respectively. These values achieved for cellulase and xylanase activity represent 12-25 fold and 36 fold higher values than the maximum so far reported for other strains of G. applanatum, respectively.

  6. Isolation, Production, and Characterization of Thermotolerant Xylanase from Solvent Tolerant Bacillus vallismortis RSPP-15

    Directory of Open Access Journals (Sweden)

    Rajeeva Gaur

    2015-01-01

    Full Text Available Sixty bacterial strains isolated from the soils sample in the presence of organic solvent were screened for xylanase production. Among them, strain RSPP-15 showed the highest xylanase activity which was identified as Bacillus vallismortis. The isolate showed maximum xylanase production (3768 U/mL in the presence of birch wood xylan and beef extract at 55°C pH 7.0 within 48 h of incubation. The enzyme activity and stability were increased 181.5, 153.7, 147.2, 133.6, and 127.9% and 138.2, 119.3, 113.9, 109, and 104.5% in the presence of Co2+, Ca2+, Mg+2, Zn+2, and Fe+3 ions (10 mM. Xylanase activity and stability were strongly inhibited in the presence of Hg and Cu ions. The enzyme was also stable in the presence of 30% of n-dodecane, isooctane, n-decane, xylene, toluene, n-hexane, n-butanol, and cyclohexane, respectively. The presence of benzene, methanol, and ethanol marginally reduced the xylanase stability, respectively. This isolate may be useful in several industrial applications owing to its thermotolerant and organic solvent resistance characteristics.

  7. Physiological and biochemical characteristics of laboratory induced mutants of Botrytis cinerea with resistance to fluazinam.

    Science.gov (United States)

    Shao, Wenyong; Zhang, Yu; Ren, Weichao; Chen, Changjun

    2015-01-01

    Botrytis cinerea is a necrotrophic and filamentous fungus with a high risk of developing resistance to fungicides. The pyridinamine fungicide fluazinam has been reported to have excellent activity against B. cinerea and better effect on controlling gray mold. In this study, the physiological and biochemical characteristics of laboratory-induced mutants of B. cinerea with resistance to fluazinam has been investigated. Compared to the wild-type strains, the fluazinam-resistant mutants had a significant decrease in respiratory rate, glycerol, oxalate, and ATP contents, and an increase in ATPase activity and sensitivity to osmotic pressure, but did not differ in cell membrane permeability. Sequencing indicated that two parental strains and four resistant mutants were identical in the nucleotide sequence of F-ATPase gene. These results will enrich our understanding of the resistance mechanism of B. cinerea to fluazinam.

  8. Effects of ozone on the sporulation, germination, and pathogenicity of Botrytis cinerea

    Energy Technology Data Exchange (ETDEWEB)

    Krause, C.R.; Weidensaul, T.C.

    1978-02-01

    Studies were initiated to determine if Botrytis cinerea conidia remain viable when grown in vivo and in vitro in the presence of ambient ozone levels and whether ozonized conidia retain pathogenicity. Experimental materials and methods used are described.

  9. Nosocomial pneumonia caused by a glucose-metabolizing strain of Neisseria cinerea.

    Science.gov (United States)

    Boyce, J M; Taylor, M R; Mitchell, E B; Knapp, J S

    1985-01-01

    We describe what appears to be the first reported case of nosocomial pneumonia caused by Neisseria cinerea. The isolate metabolized glucose when tested in BACTEC Neisseria Differentiation Kits (Johnston Laboratories), but did not produce detectable acid in cystine-Trypticase (BBL Microbiology Systems) agar medium or in modified oxidation-fermentation medium. Clinical laboratories that rely on the BACTEC method for differentiation of pathogenic neisseriae should be aware of the fact that N. cinerea may mimic N. gonorrhoeae when tested in BACTEC Neisseria Differentiation kits. The ability of N. cinerea to grow well on tryptic soy and Mueller-Hinton agars and its inability to grow on modified Thayer-Martin medium are characteristics which help to distinguish N. cinerea from N. gonorrhoeae.

  10. Identification of a new restriction endonuclease R.NciII, from Neisseria cinerea.

    Science.gov (United States)

    Piekarowicz, A

    1994-01-01

    Site-specific restriction endonuclease R. Nci II has been purified from Neisseria cinerea strain 32615. The enzyme recognizes the sequence 5' GATC 3' and its activity is inhibited by the presence of methylated adenine residue within the recognition sequence.

  11. Production of 14C-labeled gas in BACTEC Neisseria Differentiation kits by Neisseria cinerea.

    OpenAIRE

    Boyce, J M; Mitchell, E B; Knapp, J S; Buttke, T M

    1985-01-01

    Six strains of Neisseria cinerea were tested in BACTEC Neisseria Differentiation kits (Johnston Laboratories, Inc., Towson, Md.), and all yielded positive glucose growth indices and negative maltose and fructose growth indices. These results were similar to those achieved with Neisseria gonorrhoeae. However, most of the N. cinerea isolates tested yielded 3-h glucose growth indices that were lower than those obtained with gonococci. 14C-labeled gas was produced significantly faster (P less tha...

  12. Recurrent bacterial peritonitis caused by Neisseria cinerea in a chronic ambulatory peritoneal dialysis (CAPD) patient.

    Science.gov (United States)

    George, M J; DeBin, J A; Preston, K E; Chiu, C; Haqqie, S S

    1996-10-01

    We present an unusual case of recurrent (chronic ambulatory peritoneal dialysis) CAPD-associated peritonitis caused by Neisseria cinerea. Using DNA restriction fragment length polymorphism (RFLP) analysis, we determined that the recurrent infection was caused by reinfection with a different N. cinerea strain rather than relapse with the index strain and that the probable origin of the reinfecting organism was the patient's upper respiratory tract.

  13. Isolation and properties of Aspergillus niger IBT-90 xylanase for bakery.

    Science.gov (United States)

    Romanowska, Irena; Polak, Jacek; Bielecki, Stanisław

    2006-02-01

    Xylanase of low molecular weight (K II) was isolated from the fungus Aspergillus niger IBT-90 cultivated in medium with wheat bran. K II was purified by precipitation with ammonium sulphate (20-80% saturation) and gel filtration on Biogel P-10. This enzyme is most active in hydrolysis of birchwood xylan at 50 degrees C and pH 5.5. Xylanase K II has an ability to degrade 1,4-beta-bonds and to debranch substrates. It degrades not only xylans but also cellulose, an important factor for its application in bakery. Ag+, Fe3+ and NBS are strong inhibitors of the enzyme. DTT and Na+ activate xylanase K II by 24 and 13%, respectively. Enzyme K II used as additive to flour improves dough properties, increases the volume of wheat-rye and whole meal bread, and increases the porosity of crumb and the moisture of the final product, consequently extending the shelf life of bread.

  14. Study on Screening and Cultivation Conditions of Xylanase-Producing Alkalophilic Bacterial

    Institute of Scientific and Technical Information of China (English)

    Han Xiao-fang; Zheng Lian-shuang; Xie Yi-min

    2004-01-01

    An xylanase producting alkalophilic Bacillus NT-9 was obtaind by the screening method of transparent zone on the selective medium, and the effects of carbon source and nitrogen source on xylanase production were studied. The medium composed of xylose 1.5%, (NH4)2SO4 0.25%, K2HPO4 0.1%, MgSO4·7H2O 0.02%, with the initial pH of 10, was suggested to be optimal for the enzyme production in this study. When cultivatied at 37 ℃ for 72 h, the enzyme activity elaborated by the strain may reach as high as 10.5 U/mL. The xylanase produced by Bacillus NT-9 was a constituent enzyme.

  15. Characterization and high expression of recombinant Ustilago maydis xylanase in Pichia pastoris.

    Science.gov (United States)

    Han, Hongjuan; You, Shuang; Zhu, Bo; Fu, Xiaoyan; Sun, Baihui; Qiu, Jin; Yu, Chengye; Chen, Lei; Peng, Rihe; Yao, Quanhong

    2015-03-01

    A recombinant xylanase gene (rxynUMB) from Ustilago maydis 521 was expressed in Pichia pastoris, and the enzyme was purified and characterized. Phylogenetic analysis demonstrated that rxynUMB belongs to glycosyl hydrolase family 11. The Trp84, Trp95, Glu93, and Glu189 residues are proposed to be present at the active site. The apparent molecular mass of the recombinant xylananse was approximately 24 kDa, and the optimum pH and temperature were 4.3 and 50 °C, respectively. Xylanase activity was enhanced by 166 and 115% with Fe(2+) and Mn(2+), respectively. The biochemical properties of this recombinant xylanase suggest that it may be a useful candidate for a variety of commercial applications.

  16. Resistance of Malus domestica fruit to Botrytis cinerea depends on endogenous ethylene biosynthesis.

    Science.gov (United States)

    Akagi, Aya; Dandekar, Abhaya M; Stotz, Henrik U

    2011-11-01

    The plant hormone ethylene regulates fruit ripening, other developmental processes, and a subset of defense responses. Here, we show that 1-aminocyclopropane-1-carboxylic acid synthase (ACS)-silenced apple (Malus domestica) fruit that express a sense construct of ACS were more susceptible to Botrytis cinerea than untransformed apple, demonstrating that ethylene strengthens fruit resistance to B. cinerea infection. Because ethylene response factors (ERFs) are known to contribute to resistance against B. cinerea via the ethylene-signaling pathway, we cloned four ERF cDNAs from fruit of M. domestica: MdERF3, -4, -5, and -6. Expression of all four MdERF mRNAs was ethylene dependent and induced by wounding or by B. cinerea infection. B. cinerea infection suppressed rapid induction of wound-related MdERF expression. MdERF3 was the only mRNA induced by wounding and B. cinerea infection in ACS-suppressed apple fruit, although its induction was reduced compared with wild-type apple. Promoter regions of all four MdERF genes were cloned and putative cis-elements were identified in each promoter. Transient expression of MdERF3 in tobacco increased expression of the GCC-box containing gene chitinase 48.

  17. The effect of harpin on shelf life of peppers inoculated with Botrytis cinerea.

    Science.gov (United States)

    Tezcan, Himmet; Akbudak, Nuray; Akbudak, Bulent

    2013-12-01

    The preservation methods as an alternative to chemical control to prevent postharvest quality losses of peppers were examined. The efficacy of harpin treatments on peppers (Capsicum annuum L. cvs. 'Demre', 'Yalova Charleston' and 'Sari Sivri') was tested in the same conditions in two different years. Peppers grown in greenhouse were applied with four treatments consisting of harpin, Botrytis cinerea, harpin+B. cinerea and control. The harpin in B. cinerea treatments reduced the percentage of rotten fruit in cv. 'Demre' from 42.68% to 22.85%, in cv. 'Yalova Charleston' from 60.87% to 26.59% and in cv. 'Sari Sivri' from 32.83% to 12.82%. The harpin and harpin+B. cinerea peppers had a better overall appearance at the end of shelf-life. Changes in percentage of red fruit and fruit color at the end of shelf life proceeded more slowly in the harpin treated fruit. The treatments of harpin gave the best results in all three cultivars. Moreover, the values obtained from fruits subjected to harpin+B. cinerea were better than those of the fruits picked from the plants only subjected to B. cinerea. In the trials, harpin slowed down the changes leading to quality loss in fruits, in all cultivars. Thus, the positive effect of harpin was revealed more clearly especially in the fruits picked from the inoculated plants.

  18. Interactions involving ozone, Botrytis cinerea, and B. squamosa on onion leaves

    Energy Technology Data Exchange (ETDEWEB)

    Rist, D.L.

    1983-01-01

    Interactions involving Botrytis cinerea Pers., B. squamosa Walker, and ozone on onion (alium cepae L.) were investigated. Initially, threshold dosages of ozone required to predispose onion leaves to greater infection by B. cinerea and B. squamosa were determined under controlled conditions in an ozone-exposure chamber. Subsequent experiments supported the hypothesis that nutrients leaking out of ozone-injured cells stimulated lesion production by B. cinerea. The electrical conductivity of, and carbohydrate concentration in, dew collected from leaves of onion plants which had been exposed to ozone were greater than the electrical conductivity of, and carbohydrate concentration in, dew collected from leaves of other, non-exposed onion plants. When conidia of B. cinerea were suspended in dew collected from leaves of plants which had been exposed to ozone and the resulting suspension atomized onto leaves of non-exposed plants, more lesions were induced than on leaves of other non-exposed plants inoculated with conidia suspended in dew collected from plants which had not been exposed to ozone. EDU protected onion leaves from ozone-induced predisposition to these fungi under controlled conditions. Experiments designed to detect interaction between B. cinerea and B. squamosa in onion leaf blighting indicated that such interaction did not occur. Leaves were blighted rapidly when inoculated with B. squamosa whether B. cinerea was present or absent.

  19. Effectiveness of Different Classes of Fungicides on Botrytis cinerea Causing Gray Mold on Fruit and Vegetables

    Directory of Open Access Journals (Sweden)

    Joon-Oh Kim

    2016-12-01

    Full Text Available Botrytis cinerea is a necrotrophic pathogen causing a major problem in the export and post-harvest of strawberries. Inappropriate use of fungicides leads to resistance among fungal pathogens. Therefore, it is necessary to evaluate the sensitivity of B. cinerea to various classes of fungicide and to determine the effectiveness of different concentrations of commonly used fungicides. We thus evaluated the effectiveness of six classes of fungicide in inhibiting the growth and development of this pathogen, namely, fludioxonil, iprodione, pyrimethanil, tebuconazole, fenpyrazamine, and boscalid. Fludioxonil was the most effective (EC₅₀ < 0.1 μg/ml, and pyrimethanil was the least effective (EC₅₀ = 50 μg/ml, at inhibiting the mycelial growth of B. cinerea. Fenpyrazamine and pyrimethanil showed relatively low effectiveness in inhibiting the germination and conidial production of B. cinerea. Our results are useful for the management of B. cinerea and as a basis for monitoring the sensitivity of B. cinerea strains to fungicides.

  20. Large-Scale Transcriptome Analysis of Cucumber and Botrytis cinerea during Infection.

    Directory of Open Access Journals (Sweden)

    Weiwen Kong

    Full Text Available Cucumber gray mold caused by Botrytis cinerea is considered one of the most serious cucumber diseases. With the advent of Hi-seq technology, it is possible to study the plant-pathogen interaction at the transcriptome level. To the best of our knowledge, this is the first application of RNA-seq to identify cucumber and B. cinerea differentially expressed genes (DEGs before and after the plant-pathogen interaction. In total, 248,908,688 raw reads were generated; after removing low-quality reads and those containing adapter and poly-N, 238,341,648 clean reads remained to map the reference genome. There were 3,512 cucumber DEGs and 1,735 B. cinerea DEGs. GO enrichment and KEGG enrichment analysis were performed on these DEGs to study the interaction between cucumber and B. cinerea. To verify the reliability and accuracy of our transcriptome data, 5 cucumber DEGs and 5 B. cinerea DEGs were chosen for RT-PCR verification. This is the first systematic transcriptome analysis of components related to the B. cinerea-cucumber interaction. Functional genes and putative pathways identified herein will increase our understanding of the mechanism of the pathogen-host interaction.

  1. Improvement of xylanase production by Penicillium canescens 10-10c in solid-state fermentation

    Directory of Open Access Journals (Sweden)

    Thonart P.

    2008-01-01

    Full Text Available Among hemicellulases, xylanases are catalysts of considerable interest so as fundamental than applied point of view. However,it is paradoxical to note that the high cost of their production limits their use on a large scale. The use of purified xylan as culturesubstrate increases the production cost of the enzyme. Consequently, for commercial applications, it is advisable to developprocesses starting from inexpensive substrates. The purpose of this study is to optimise xylanases production in solid-statefermentation based on agricultural residues. The strain is Penicillium canescens 10-10c, selected in our laboratory for his abilityto produce xylanase activity free of cellulase. Assays concern optimization of different culture parameters in order to developin the future a solid-state fermentation reactor with soya oil cake. These parameters are: medium composition, temperatureincubation, induction and repression mechanisms. Soya oil cake in pellets (size > 10 mm gave a higher enzymatic activity.Great volume of culture medium reduced the enzymatic production. The presence of lactose, saccharose or starch of corn hasa positive effect on the production of xylanase while the presence of xylose, mannose, galactose, arabinose, cellobiose andpectin or methylcellulose reduces the production of xylanase. The sources of phosphorus (di-potassic and di-sodic enhancexylanase production. The enzymatic production obtained in Erlenmeyer flasks (250 ml after 7 days incubation at 30°C isabout 14 000 U.g-1 of carbon source. The nature of inoculum affects the enzymatic productivity. Indeed, better productivitywas obtained with inoculation by solid preculture (956 U.g-1 per day than liquid preculture (473 U.g-1 per day and sporessuspension (383 U.g-1 per day. These observed enzymatic activity levels are higher than those related in the literature, whichshows all the potentialities of this strain and this technique for the production of xylanase and allow to develop

  2. Exploration of Potential Actinomycetes from CIFOR Forest Origin as Antimicrobial, Antifungus, and Producing Extracellular Xylanase

    Directory of Open Access Journals (Sweden)

    Sipriyadi Sipriyadi

    2016-03-01

    Full Text Available This study aimed to isolate and explore the actinomycetes of CIFOR forest origin as an antimicrobial and antifungal agent, to produce an extracellular xylanase, and to identify isolates based on 16S rRNA gene sequences. Actinomycetes were isolated using Humic-acid Vitamin-B agar (HV media. Actinomycetes colonies that grow on the medium HV was subsequently purified by growing them on yeast malt agar (YMA media, then an antagonistic test of selected bacteria against Bacillus sp., Escherichia coli, Fusarium oxysporum, and Sclerotium sp was performed. Xylanase activity test was detected by observing a clear zone, followed by identification. Total of 35 isolates of actinomycetes isolated based on their colony morphology characteristics and diverse types of spore chains showed Streptomyces spp. of isolates CFR-06, CFR-15, CFR-17, CFR-18, and CFR-19 were able to inhibit the growth of Bacillus sp.. The highest inhibition zone has a diameter of 10.1 mm (isolate CFR-17. Isolates CFR-01 and CFR-15 were able to inhibit the growth of E. coli with the highest inhibition zone diameter of 5.1 mm (isolate CFR-15. Isolates CFR-29 and CFR-12 were able to inhibit the growth of F. oxysporum while isolate CFR-35 were able to inhibit the growth of Sclerotium sp.. Xylanase activity test showed that isolates CFR-12, CFR-20, CFR-22, CFR-24, CFR-25, CFR-30, CFR-33, CFR-34 have an ability to produce extracellular xylanase enzyme. Actinomycetes isolate (Xyl_22 as a potential xylanase enzyme producer was closely related with Streptomyces drozdowicii by the maximum similarity of 99%.How to CiteSipriyadi, S., Lestari, Y., Wahyudi, A., Meryandini, A., & Suhartono, M. T. (2016. Exploration Potential CIFOR Forest actinomycetes origin as Antimicrobial, Anti Fungus and Producing Enzymes Extracellular Xylanase. Biosaintifika: Journal of Biology & Biology Education, 8(1, 94-102.

  3. Bioprocess and biotecnology: effect of xylanase from Aspergillus niger and Aspergillus flavus on pulp biobleaching and enzyme production using agroindustrial residues as substract

    OpenAIRE

    de Alencar Guimaraes, Nelciele ; Sorgatto, Michele ; Nogueira, Simone de Peixoto; Betini, Jorge Henrique ; Zanoelo, Fabiana ; Marques, Maria ; Polizeli, Maria de Lourdes Teixeira de Moraes; Giannesi, Giovana C

    2013-01-01

    This study compares two xylanases produced by filamentous fungi such as A. niger and A. flavus using agroindustrial residues as substract and evaluated the effect of these enzymes on cellulose pulp biobleaching process. Wheat bran was the best carbon source for xylanase production by A. niger and A. flavus. The production of xylanase was 18 and 21% higher on wheat bran when we compare the xylanase production with xylan. At 50°C, the xylanase of A. niger retained over 85% activity with 2 h of ...

  4. Difficulties in differentiating Neisseria cinerea from Neisseria gonorrhoeae in rapid systems used for identifying pathogenic Neisseria species.

    OpenAIRE

    Boyce, J M; Mitchell, E B

    1985-01-01

    Neisseria cinerea and Neisseria gonorrhoeae may occur at the same body sites and may have similar colony morphologies. Ideally, systems used for rapid identification of N. gonorrhoeae should be able to differentiate N. cinerea from gonococci. We tested seven N. cinerea strains using the Gonochek II (Du Pont Diagnostics), Minitek (BBL Microbiology Systems), RapID-NH (Innovative Diagnostics, Inc.), RIM-N (American Microscan), and Phadebact (Pharmacia Diagnostics) systems. We found that the reac...

  5. Stable expression of a thermostable xylanase of Clostridium thermocellum in cultured tobacco cells.

    Science.gov (United States)

    Kimura, Tetsuya; Mizutani, Tomomi; Sakka, Kazuo; Ohmiya, Kunio

    2003-01-01

    Two distinct domains of the xynA gene from Clostridium thermocellum encoding a xylanase catalytic domain (XynAl) and a xylanase catalytic domain with a cellulose binding domain (XynA2) under the control of the cauliflower mosaic virus 35S promoter were electroporated into cultured tobacco BY-2 cells. Transgenic BY -2 calli expressing xylan-hydrolyzing activity were obtained at high frequency for both genes. Western blot analysis using an anti-XynA antibody indicated that XynAl and XynA2 were produced in these calli.

  6. Purification and characterization of five cellulases and one xylanase from Penicillium brasilianum IBT 20888

    DEFF Research Database (Denmark)

    Jørgensen, Henning; Eriksson, T.; Borjesson, J.;

    2003-01-01

    The filamentous fungus Penicillium brasilianum IBT 20888 was cultivated on a mixture of 30 g l(-1) cellulose and 10 g l(-1) xylan for 111 h and the resulting culture filtrate was used for protein purification. From the cultivation broth, five cellulases and one xylanase were purified. Hydrolysis...... the cellulose-binding domain or an essential part of it. The basic xylanase (pI > 9) was only active towards xylan. Two of the purified cellulases with endoglucanase activity were partly sequenced and based on sequence homology with known enzymes they were classified as belonging to families 5 and 12...

  7. PRODUCTION OF SINGLE CELL PROTEIN, ESSENTIAL AMINO ACIDS, AND XYLANASE BY PENICILLIUM JANTHINELLUM

    Directory of Open Access Journals (Sweden)

    Mala B. Rao

    2010-11-01

    Full Text Available Microbial biomass having 46% crude protein content and enriched with essential amino acids as well as extracellular xylanase activity (100-150 IU/ml was produced by an efficient fungal strain, Penicillium janthinellum (NCIM St-F-3b. Optimization studies for maximum xylanase and biomass production showed that the fungus required a simple medium containing bagasse hemicellulose as carbon source and ammonium sulphate as the nitrogen source. Therefore bagasse, which is a waste product of the sugar industry, can be efficiently used in microbioal biomass protein preparation for animal feed.

  8. Classification, mode of action and production strategy of xylanase and its application for biofuel production from water hyacinth.

    Science.gov (United States)

    Uday, Uma Shankar Prasad; Choudhury, Payel; Bandyopadhyay, Tarun Kanti; Bhunia, Biswanath

    2016-01-01

    Xylanases are classified under glycoside hydrolase families which represent one of the largest groups of commercial enzymes. Depolymerizing xylan molecules into monomeric pentose units involves the synergistic action of mainly two key enzymes which are endo-β-xylanase and β-xylosidase. Xylanases are different with respect to their mode of action, substrate specificities, biochemical properties, 3D structure and are widely produced by a spectrum of bacteria and fungi. Currently, large scale production of xylanase can be produced through the application of genetic engineering tool which allow fast identification of novel xylanase genes and their genetic variations makes it an ideal enzymes. Due to depletion of fossil fuel, there is urgent need to find out environment friendly and sustainable energy sources. Therefore, utilisation of cheap lignocellulosic materials along with proper optimisation of process is most important for cost efficient ethanol production. Among, various types of lignocellulosic substances, water hyacinth, a noxious aquatic weed, has been found in many tropical. Therefore, the technological development for biofuel production from water hyacinth is becoming commercially worthwhile. In this review, the classification and mode of action of xylanase including genetic regulation and strategy for robust xylanase production have been critically discussed from recent reports. In addition various strategies for cost effective biofuel production from water hyacinth including chimeric proteins design has also been critically evaluated.

  9. Cloning, expression and applicability of thermo-alkali-stable xylanase of Geobacillus thermoleovorans in generating xylooligosaccharides from agro-residues.

    Science.gov (United States)

    Verma, Digvijay; Satyanarayana, T

    2012-03-01

    A xylanase gene (xyl-gt) of 1.224 kbp was cloned from the extremely thermophilic bacterium Geobacillus thermoleovorans that encodes a protein containing 408 amino acid residues. Eight conserved regions (signature sequences) of GH family 10 xylanases have been found in the xylanase. When the xylanase gene was cloned and expressed in Escherichia coli BL21 (DE3), the recombinant strain produced xylanase titer of 270 U mg(-1) which is 27-fold higher than the wild strain. It is optimally active at 80°C and pH 8.5 with a high thermostability over broad range of pH (6-12) and temperature (40-100°C). The end products of the hydrolysis of birch wood xylan and agro-residues included xylobiose, xylotriose, xylotetraose and xylopentaose. The xylanase of G. thermoleovorans is one of the rare xylanases that exhibits thermo-alkali-stability, and thus, it is a suitable candidate for pre-bleaching of paper pulps and generating xylooligosaccharides from agro-residues for use as prebiotics.

  10. Attachment Capability of Antagonistic Yeast Rhodotorula glutinis to Botrytis cinerea Contributes to Biocontrol Efficacy.

    Science.gov (United States)

    Li, Boqiang; Peng, Huaimin; Tian, Shiping

    2016-01-01

    Rhodotorula glutinis as an antagonism show good biocontrol performance against various post-harvest diseases in fruits. In the present study, strong attachment capability of R. glutinis to spores and hyphae of Botrytis cinerea was observed. Further analysis showed that certain protein components on the yeast cell surface played critical role during the interaction between R. glutinis and B. cinerea. The components mainly distributed at the poles of yeast cells and might contain glycosylation modification, as tunicamycin treated yeast cells lost attachment capability to B. cinerea. To investigate contributions of attachment capability of R. glutinis to its biocontrol efficacy, yeast cells were mutagenized with 3% methane-sulfonic acid ethyl ester (EMS), and a mutant CE4 with stable non-attaching phenotype was obtained. No significant difference was found on colony, cell morphology, reproductive ability, and capsule formation between the mutant and wild-type. However, there was a distinct difference in India ink positive staining patterns between the two strains. Moreover, wild-type strain of R. glutinis showed better performance on inhibiting spore germination and mycelial growth of B. cinerea than CE4 strain when yeast cells and B. cinerea were co-cultured in vitro. In biocontrol assay, both wild-type and CE4 strains showed significant biocontrol efficacy against gray mold caused by B. cinerea in apple fruit, whereas, control effect of CE4 strain was lower than that of wild-type. Our findings provided new evidences that attachment capability of R. glutinis to B. cinerea contributed to its biocontrol efficacy.

  11. Evaluation of fast volatile analysis for detection of Botrytis cinerea infections in strawberry.

    Science.gov (United States)

    Vandendriessche, Thomas; Keulemans, Johan; Geeraerd, Annemie; Nicolai, Bart M; Hertog, Maarten L A T M

    2012-12-01

    Grey mold (Botrytis cinerea) is one of the major phytopathogens causing serious losses during strawberry postharvest and storage. B. cinerea-host interaction affect emissions of volatile compounds during infection resulting in a characteristic earthy, mushroom odor. Therefore, the objective of this study was to evaluate two analytical techniques based on fast volatile analysis on their performance for monitoring evolution and early detection of B. cinerea infections in strawberry. In a first experiment headspace multi-capillary column-ion mobility spectrometry (HS MCC-IMS) has been successfully used to evaluate development of strawberry aroma during shelflife. In a second experiment the same technique has been used to detect the degree of B. cinerea infection through changes in the volatile profile. Additionally, these samples were analyzed with headspace solid-phase-microextraction fast GC-MS (HS SPME fast GC-MS). Both HS MCC-IMS and HS SPME fast GC-MS could determine the changes in volatile composition as a function of the degree of B. cinerea infection as determined by an enzyme-linked immunosorbent assay (ELISA) and could be used to follow the evolution of infection. According to the ELISA data, some fruit were infected even without any symptoms and volatiles produced by the fungus may be overshadowed by the fruit volatiles. Therefore, both analytical techniques could not be used for early detection of B. cinerea infections. After identification of the volatile compounds and multivariate data analysis, potential biomarkers specific for B. cinerea were highlighted, being 3-methylbutanal, cis-4-decenal, 2-methyl-1-butanol, 2-methyl-1-propanol, 1-octen-3-one and 1-octen-3-ol.

  12. Functional analysis of glycoside hydrolase family 8 xylanases shows narrow but distinct substrate specificities and biotechnological potential.

    Science.gov (United States)

    Pollet, Annick; Schoepe, Jan; Dornez, Emmie; Strelkov, Sergei V; Delcour, Jan A; Courtin, Christophe M

    2010-08-01

    The potential of glycoside hydrolase family (GH) 8 xylanases in biotechnological applications is virtually unexplored. Therefore, the substrate preference and hydrolysis product profiles of two GH8 xylanases were evaluated to investigate their activities and substrate specificities. A GH8 xylanase from an uncultured bacterium (rXyn8) shows endo action but very selectively releases xylotriose from its substrates. It has a higher activity than the Pseudoalteromonas haloplanktis GH8 endo-xylanase (PhXyl) on xylononaose and smaller xylo-oligosaccharides. PhXyl preferably degrades xylan substrates with a high degree of polymerization. It is sterically more hindered by arabinose substituents than rXyn8, producing larger end hydrolysis products. The specificities of rXyn8 and PhXyl differ completely from these of the previously described GH8 xylanases from Bifidobacterium adolescentis (BaRexA) and Bacillus halodurans (BhRex). As reducing-end xylose-releasing exo-oligoxylanases, they selectively release xylose from the reducing end of small xylo-oligosaccharides. The findings of this study show that GH8 xylanases have a narrow substrate specificity, but also one that strongly varies between family members and is distinct from that of GH10 and GH11 xylanases. Structural comparison of rXyn8, PhXyl, BaRexA, and BhRex showed that subtle amino acid changes in the glycon as well as the aglycon subsites probably form the basis of the observed differences between GH8 xylanases. GH8 xylanases, therefore, are an interesting group of enzymes, with potential towards engineering and applications.

  13. Cloning, sequencing and expression of a xylanase gene from the maize pathogen Helminthosporium turcicum

    DEFF Research Database (Denmark)

    Degefu, Y.; Paulin, L.; Lübeck, Peter Stephensen

    2001-01-01

    A gene encoding an endoxylanase from the phytopathogenic fungus Helminthosporium turcicum Pass. was cloned and sequenced. The entire nucleotide sequence of a 1991 bp genomic fragment containing an endoxylanase gene was determined. The xylanase gene of 795 bp, interrupted by two introns of 52 and ...

  14. The Production of Fungal Mannanase, Cellulase and Xylanase Using Palm Kernel Meal as a Substrate

    Directory of Open Access Journals (Sweden)

    Nisa SAE-LEE

    2007-01-01

    Full Text Available Extracellular enzymes including mannanase, cellulase and xylanase from Aspergillus wentii TISTR 3075, Aspergillus niger, Aspergillus oryzae, Trichoderma reesei TISTR 3080 and Penicillium sp. were investigated. The enzymes were produced in solid-state fermentation using palm kernel meal (PKM as a substrate. All fungal strains produced mainly mannanase. A maximum activity of 24.9 U/g koji was observed in A. wentii TISTR 3075 with a specific activity of 1.5 U/mg protein. During PKM fermentation, there was also found low concomitantly of cellulase and xylanase activities with high mannanase activity in all strains. The degradation of non-starch polysaccharides (NSPs in PKM by these fungal strains was indicated by the increased mannanase, cellulase and xylanase activities which correlated with the increase in reducing sugar content and pH profiles during PKM fermentation. PKM was shown to be more suitable for production of mannanase than cellulase and xylanase for all strains because of the high content of mannan as an inducer in PKM. Increases in enzyme yield might be obtained by optimizing the culture conditions.

  15. Xylanase production by the thermophilic mold Humicola lanuginosa in solid-state fermentation.

    Science.gov (United States)

    Kamra, Pankaj; Satyanarayana, T

    2004-11-01

    Among the lignocellulosic substrates tested, wheat bran supported a high xylanase (EC 3.2.1.8) secretion by Humicola lanuginosa in solid-state fermentation (SSF). Enzyme production reached a peak in 72 h followed by a decline thereafter. Enzyme production was very high (7832 U/g of dry moldy bran) when wheat bran was moistened with tap water at a substrate--to--moistening agent ratio of 1:2.5 (w/v) and an inoculum level of 3 x 106 spores/10 g of wheat bran at a water activity (aw) of 0.95. Cultivation of the mold in large enamel trays yielded a xylanase titer comparable with that in flasks. Parametric optimization resulted in a 31% increase in enzyme production in SSF. Xylanase production was approx 23-fold higher in SSF than in submerged fermentation (SmF). A threshold constitutive level of xylanase was secreted by H. lanuginosa in a medium containing glucose as the sole carbon source. The enzyme was induced by xylose and xylan. Enzyme synthesis was repressed beyond 1.0% (w/v) xylose in SmF, whereas it was unaffected up to 3.0% (w/w) in SSF, suggesting a minimization of catabolite repression in SSF.

  16. Improvement of the quality of wheat bread by addition of glycoside hydrolase family 10 xylanases.

    Science.gov (United States)

    Zheng, Han; Guo, Bing; Chen, Xiu-Lan; Fan, Sou-Jin; Zhang, Yu-Zhong

    2011-04-01

    Although many xylanases are widely used in the baking industry, only one glycoside hydrolase family 10 (GH 10) xylanase has previously been reported to be effective in baking. In this study, we compared the effectiveness of two GH 10 xylanases, psychrophilic XynA from Glaciecola mesophila and mesophilic EX1 from Trichoderma pseudokoningii, in bread making. The optimal dosages needed to improve wheat flour dough and bread quality were 270-U/kg flour for EX1 and 0.9-U/kg flour for XynA. At their optimal dosage, both XynA and EX1 had significant dough-softening ability, resulting in a 50% reduction in Brabender units. XynA was more effective than EX1 in reducing the time to reach maximum consistency. XynA and EX1 showed similar effects in improving the bread volume (~30% increase). EX1 was more effective in reducing the initial crumb firmness. Although both enzymes exhibited similar anti-staling effects on the bread, based on a decrease in the bread firmness, XynA had a greater effect on reducing the firming rate, and EX1 showed an enhanced reduction in the initial firmness. These results show that these two GH 10 xylanases have unique advantages in improving dough and bread quality and indicate their potential in bread making.

  17. Production of crude xylanase from Thermoascus aurantiacus CBMAI 756 aiming the baking process.

    Science.gov (United States)

    Oliveira, Denise S; Meherb-Dini, Carolina; Franco, Célia M L; Gomes, Eleni; Da-Silva, Roberto

    2010-09-01

    In recent years, the baking industry has focused its attention on substituting several chemical compounds with enzymes. Enzymes that hydrolyze nonstarch polysaccharides, such as xylanase, lead to the improvement of rheological properties of dough, loaf specific volume, and crumb firmness. The purpose of this study was to find a better solid-state fermentation substrate to produce high levels of xylanase and low levels of protease and amylase, which are enzymes involved in bread quality, from Thermoascus aurantiacus CBMAI 756. Wheat bran, corncob, and corn straw were used as energy sources. The enzyme extract of corncob showed high xylanase activity (130 U/mL) and low amylase and protease activity (baking industry, because it results in a slower degradation of gluten. Our results confirm this finding, because the enzyme obtained by fermentation in corncob resulted in a gluten with a higher specific volume than all the other substrates that were tested. The crude xylanase presented maximum activity at a pH of 5, and the optimum temperature was 75 °C. It was stable up to 70 °C for an hour and at a pH range from 4 to 10.

  18. [Biosynthesis of cellulolytic enzymes and xylanase during submerged cultivation of the fungus Aspergillus terreus 17P].

    Science.gov (United States)

    Loginova, L G; Guzhova, E P; Ismanlova, D Iu; Burdenko, L G

    1978-01-01

    The fungus Aspergillus terreus 17P--producer of cellulolytic enzymes--was cultivated in the Biotec 10 l fermenter on the medium containing minced and heated (at 200 degrees) wheat straw aerated with a different rate. At the mixing rate of 350 rpm and aeration rate of 0.7 r/rpm on the fourth day the culture liquid was obtained whose filtrate contained an active complex of cellulolytic enzymes and xylanase: CI--3.4; APB--1.1, Cx--35.7, cellobiase--0.23, xylanase--73.8 units/ml. The fractionation of the culture liquid filtrate with ammonium sulphate showed that the fraction precipitated at an interval of saturation of 0.3--0.7 contained the largest portion of cellulolytic enzymes and xylanase. The isolated enzymic preparations had a cellulolytic and xylanase activity and contained lipase, pectinase, laminarinase. They also contained low quantities of amylase, protease, beta-1,4- and beta-1,6-glucanase. Enzymic hylrolysis by the Asp. terreus 17P preparation of straw yielded glucose and xylose, of cotton, Na-KMC, cellobiose--glucose, Xylane hydrolyzate contained xylose and arabinose.

  19. Thermal Stability and Thermodynamics of Xylanase from Melanocarpus albomyces in Presence of Polyols and Salts

    Directory of Open Access Journals (Sweden)

    Gupteshwar Gupta

    2014-08-01

    Full Text Available An extracellular xylanase from the thermophilic fungus Melanocarpus albomyces IIS 68 was evaluated for its activity and stability in the presence of polyols and salts at 60 °C, and found to be an effective protecting agent for thermal deactivation of enzyme. Response surface methodology was employed to study the synergistic effects of glycerol and NaCl (thermo-stabilizers for xylanase stability. The addition of these thermo-stabilizers resulted in more than a 10-fold increment in enzyme half-life. Activation energy (Ea and thermodynamic parameters such as ∆H, ∆G, and ∆S were calculated for the thermal inactivation of free and immobilized xylanase. The immobilized enzyme underwent substantially less conformational changes because of its enhanced stability and increased compactness, providing better thermo-stability at elevated temperatures. These findings suggest that the combined effect of glycerol and sodium chloride serves as a potential stabilizer for extracellular thermophilic xylanase, which finds commercial application in many industries, especially in the pulp and paper industry.

  20. Xylanases of marine fungi of potential use for biobleaching of paper pulp

    Digital Repository Service at National Institute of Oceanography (India)

    Raghukumar, C.; Muraleedharan, U.; Gaud, V.R.; Mishra, R.

    bleaching of sugarcane bagasse pulp by a 60 min treatment at 55oC, resulting in a decrease of 10 kappa numbers and a 30% reduction in consumption of chlorine during bleaching process. The culture filtrate showed peaks of xylanase activity at acidic pH (3...

  1. Characterization of a bifunctional xylanase/endoglucanase from yak rumen microorganisms.

    Science.gov (United States)

    Chang, Lei; Ding, Mozhu; Bao, Lei; Chen, Yingzhi; Zhou, Jungang; Lu, Hong

    2011-06-01

    A new gene, RuCelA, encoding a bifunctional xylanase/endoglucanase, was cloned from a metagenomic library of yak rumen microorganisms. RuCelA showed activity against xylan and carboxymethylcellulose (CMC), suggesting bifunctional xylanase/endoglucanase activity. The optimal conditions for xylanase and endoglucanase activities were 65°C, pH 7.0 and 50°C, pH 5.0, respectively. In addition, the presence of Co(+) and Co(2+) can greatly improve RuCelA's endoglucanase activity, while inhibits its xylanase activity. Further examination of substrate preference showed a higher activity against barley glucan and lichenin than against xylan and CMC. Using xylan and barley glucan as substrates, RuCelA displayed obvious synergistic effects with β-1,4-xylosidase and β-1,4-glucosidase. Generation of soluble oligosaccharides from lignocellulose is the key step in bioethanol production, and it is greatly notable that RuCelA can produce xylo-oligosaccharides and cello-oligosaccharides in the continuous saccharification of pretreated rice straw, which can be further degraded into fermentable sugars. Therefore, the bifunctional RuCelA distinguishes itself as an ideal candidate for industrial applications.

  2. Toevoeging van ß-glucanase en xylanase aan mengvoeders voor gespeende biggen

    NARCIS (Netherlands)

    Scholten, R.H.J.; Binnendijk, G.P.

    1997-01-01

    Op het proefbedrijf van het Proefstation voor de Varkenshouderij te Rosmalen is van november 1995 tot en met februari 1996 een onderzoek uitgevoerd om na te gaan wat de mogelijkheden zijn van toevoeging van de enzymen B-glucanase en xylanase (PorzymeSlOO@) aan biggenrantsoenen

  3. Marine-derived fungus Aspergillus cf. tubingensis LAMAI 31: a new genetic resource for xylanase production.

    Science.gov (United States)

    Dos Santos, Juliana A; Vieira, Juliana M F; Videira, Alexandre; Meirelles, Lucas A; Rodrigues, André; Taniwaki, Marta H; Sette, Lara D

    2016-03-01

    Marine-derived fungi have been reported as relevant producers of enzymes, which can have different properties in comparison with their terrestrial counterparts. The aim of the present study was to select from a collection of 493 marine-derived fungi the best producer of xylanase in order to evaluate the enzymatic production under different conditions. A total of 112 isolates produced xylanase in solid medium containing xylan as the carbon source, with 31 of them able to produce at least 10 U/mL of the enzyme. The best production (49.41 U/mL) was achieved by the strain LAMAI 31, identified as Aspergillus cf. tubingensis. After confirming the lack of pathogenicity (absence of ochratoxin A and fumonisin B2 production) this fungus was submitted to the experimental design in order to evaluate the effect of different variables on the enzymatic production, with the aim of optimizing culture conditions. Three experimental designs (two Plackett-Burman and one factorial fractional) were applied. The best condition for the enzymatic production was defined, resulting in an increase of 12.7 times in comparison with the initial production during the screening experiments. In the validation assay, the peak of xylanase production (561.59 U/mL) was obtained after 96 h of incubation, being the best specific activity achieved after 72 h of incubation. Xylanase from A. cf. tubingensis LAMAI 31 had optimum pH and temperature at 5.0 and 55 °C, respectively, and was shown to be stable at a range of 40-50 °C, and in pH from 3.6 to 7.0. Results from the present work indicate that A. cf. tubingensis LAMAI 31 can be considered as a new genetic resource for xylanase production.

  4. Domain-swapping of mesophilic xylanase with hyper-thermophilic glucanase

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    Liu Liangwei

    2012-06-01

    Full Text Available Abstract Background Domain fusion is limited at enzyme one terminus. The issue was explored by swapping a mesophilic Aspergillus niger GH11 xylanase (Xyn with a hyper-thermophilic Thermotoga maritima glucanase (Glu to construct two chimeras, Xyn-Glu and Glu-Xyn, with an intention to create thermostable xylanase containing glucanase activity. Results When expressed in E. coli BL21(DE3, the two chimeras exhibited bi-functional activities of xylanase and glucanase. The Xyn-Glu Xyn moiety had optimal reaction temperature (Topt at 50 °C and thermal in-activation half-life (t1/2 at 50 °C for 47.6 min, compared to 47 °C and 17.6 min for the Xyn. The Glu-Xyn Xyn moiety had equivalent Topt to and shorter t1/2 (5.2 min than the Xyn. Both chimera Glu moieties were more thermostable than the Glu, and the three enzyme Topt values were higher than 96 °C. The Glu-Xyn Glu moiety optimal pH was 5.8, compared to 3.8 for the Xyn-Glu Glu moiety and the Glu. Both chimera two moieties cooperated with each other in degrading substrates. Conclusions Domain-swapping created different effects on each moiety properties. Fusing the Glu domain at C-terminus increased the xylanase thermostability, but fusing the Glu domain at N-terminus decreased the xylanase thermostability. Fusing the Xyn domain at either terminus increased the glucanase thermostability, and fusing the Xyn domain at C-terminus shifted the glucanase pH property 2 units higher towards alkaline environments. Fusing a domain at C-terminus contributes more to enzyme catalytic activity; whereas, fusing a bigger domain at N-terminus disturbs enzyme substrate binding affinity.

  5. Xylanase production by Aspergillus awamori under solid state fermentation conditions on tomato pomace

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    Marcelo A. Umsza-Guez

    2011-12-01

    Full Text Available In this work, tomato pomace, a waste abundantly available in the Mediterranean and other temperate climates agro-food industries, has been used as raw material for the production of some hydrolytic enzymes, including xylanase, exo-polygalacturonase (exo-PG, cellulase (CMCase and α-amylase. The principal step of the process is the solid state fermentation (SSF of this residue by Aspergillus awamori. In several laboratory experiments, maximum xylanase and exo-PG activities were measured during the first days of culture, reaching values around 100 and 80 IU/gds (international units of enzyme activity per gram of dried solid, respectively. For CMCase and α-amylase production remained almost constant along fermentation, with average values of 19 and 21.5 IU/gds, respectively. Experiments carried out in a plate-type bioreactor at lab scale showed a clear positive effect of aeration on xylanase and CMCase, while the opposite was observed for exo-PG and α-amylase. In general, xylanase was the enzyme produced in higher levels, thus the optimum conditions for the determination of the enzyme activity was characterized. The xylanase activity shows an optimum pH of 5 and an optimum temperature of 50 ºC. The enzyme is activated by Mg2+, but strongly inhibited by Hg2+ and Cu2+. The enzymatic activity remains quite high if the extract is preserved in a range of pH from 3 to 10 and a temperature between 30 ºC to 40 ºC.

  6. Screening of xylanase activity of Streptomyces albidoflavus PSM-3n isolated from Uttarakhand

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    Pushpendra Sharma

    2013-07-01

    Full Text Available Background: Awareness towards the environmental pollution had made the evolution of green technology by which enzymes got special attention in industries. The enzymes replaced chemical catalysts in manufacturing various chemicals, agricultural and pharmaceutical products. Material and methods: Actinomycetes were isolated and screened for their ability to produce xylanase. For the most promising isolate, selection of media, effect of pH, temperature, metal ions, and detergents on enzyme production and activity was studied. Results: Out of 29 isolates, 22 isolates showed xylanase activity. Out of 22 xylanase producing isolate, 05 isolates were selected for secondary screening on the basis of their clear zone size. The most promising isolate PSM-3n was identified as Streptomyces albidoflavus. It produces maximum enzyme (xylanase in media Horikoshi and Ikura having carbon and nitrogen sources as oat meal and urea respectively. The optimum pH and temperature for the enzyme production was 4.0 and 45°C respectively. The enzyme activity was found maximum at temperature 50°C and enhanced in the presence of Fe3+ ions. There was a reduction in the enzyme activity in the presence of detergents like SDS, tween-20 and tween-80. The enzyme was fairly stable at 50°C for 1 h. Conclusion: The enzyme produced by the isolate PSM-3n is fairly heat stable and highly acid stable. The activity of the enzyme was increased in presence of Fe3+ ions while decreased in presence of SDS. Therefore, further studies are required for purification of xylanase for its application potential in pulp bioleaching processes and in the functional food industry.

  7. A comparison of plate assay methods for detecting extracellular cellulase and xylanase activity.

    Science.gov (United States)

    Meddeb-Mouelhi, Fatma; Moisan, Jessica Kelly; Beauregard, Marc

    2014-11-01

    Identification of microorganisms for the production of carbohydrolytic enzymes is extremely important given the increased demand for these enzymes in many industries. To this end, dye-polysaccharide interactions which provide a visual indication of polymer hydrolysis (clear zones or halos) have been used for decades. For the detection of extracellular cellulase or xylanase activity many laboratories use Gram's iodine as the chromogenic dye, as it is a more rapid initial screening method compared to the use of other dyes. Here, we compared Gram's iodine and Congo red as indicators of polysaccharide hydrolysis. We attempted to detect cellulase activity using carboxymethylcellulose, and xylanase activity using birchwood xylan, in fourteen uncharacterized bacteria isolated from wood chips. Our results indicate that Gram's iodine may lead to identification of false positives in a typical screening protocol and that Congo red allows for avoidance of such pitfall. Congo red allowed detection of cellulase activity from live microbial colonies but not Gram's iodine. To confirm this, detection of enzymatic activity was also assessed using cell-free enzyme preparations. Congo red was found to be reliable in detecting cellulase activity with isolated enzymes preparations. Under the same conditions, neither of these dyes detected xylanase activity, despite independent evidence of xylanase activity for one of the preparations. We detected xylanase activity for this particular enzyme preparation using a coloured derivative of xylan (Remazol Brillant Blue R-xylan adduct) that respond to xylan hydrolysis. Our results suggest that methods that rely on interactions between a dye (Congo red or Gram's iodine) and a polymeric substrate (carboxymethylcellulose or birchwood xylan) for indirect detection of hydrolysis may require the use of relevant controls and independent confirmation of enzymatic activities.

  8. Detection transposable elements in Botrytis cinerea in latent infection stage from symptomless apples

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    Jorge G Fernández

    2014-02-01

    Full Text Available Objective: T o detect Botrytis cinerea ( B. cinerea latent infections on apples before storage, which is essential for effective control strategies in the fruit postharvest industry. Methods: I n the present study, a polymerase chain reaction detection method, based on primers designed on B. cinerea transposable elements ( boty and flipper and intergenic spacer region as internal control, were utilized to reveal the presence of symptomless infections on apple fruits. T his molecular method proved to be highly specific and sensitive in detecting latent infections. I t revealed the presence of the pathogen in 83 % of the samples from infected apples with 10 4 conidia/ m L , whereas those infected with 10 6 conidia/m L detected 94 % as compared to the traditional method that revealed the pathogen in 40 % and 66 % of the samples inoculated with 10 4 and 10 6 conidia/m L respectively. F urthermore, the method characterized B. cinerea as subpopulation transposa-type by the presence of the transposable elements boty and flipper Results: T he results obtained from DNA quantification method were compared with enzyme- linked immunosorbent assay and these studies showed good correlation. T herefore our method has important advantages compared with others detection methods for B. cinerea, because the proposed methodology allowed distinguishes between its two subpopulations ( vacuma and transposa and this would allow establish possible appropriate control strategies. Conclusions: F inally, the method can be an interesting alternative for its possible application in the phytosanitary programs of the fruit industry worldwide.

  9. Detection transposable elements in Botrytis cinerea in latent infection stage from symptomless apples

    Institute of Scientific and Technical Information of China (English)

    Jorge G Fernndez; Martn A Fernndez-Baldo; Claudio Muoz; Eloy Salinas; Julio Raba; Mara I Sanz

    2014-01-01

    Objective:To detect Botrytis cinerea (B. cinerea) latent infections on apples before storage, which is essential for effective control strategies in the fruit postharvest industry. Methods:In the present study, a polymerase chain reaction detection method, based on primers designed on B. cinerea transposable elements (boty and flipper) and intergenic spacer region as internal control, were utilized to reveal the presence of symptomless infections on apple fruits. This molecular method proved to be highly specific and sensitive in detecting latent infections. It revealed the presence of the pathogen in 83%of the samples from infected apples with 104 conidia/mL, whereas those infected with 106 conidia/mL detected 94%as compared to the traditional method that revealed the pathogen in 40%and 66%of the samples inoculated with 104 and 106 conidia/mL respectively. Furthermore, the method characterized B. cinerea as subpopulation transposa-type by the presence of the transposable elements boty and flipper Results:The results obtained from DNA quantification method were compared with enzyme-linked immunosorbent assay and these studies showed good correlation. Therefore our method has important advantages compared with others detection methods for B. cinerea, because the proposed methodology allowed distinguishes between its two subpopulations (vacuma and transposa) and this would allow establish possible appropriate control strategies. Conclusions:Finally, the method can be an interesting alternative for its possible application in the phytosanitary programs of the fruit industry worldwide.

  10. Metabolic activities of five botryticides against Botrytis cinerea examined using the Biolog FF MicroPlate.

    Science.gov (United States)

    Wang, Hancheng; Wang, Jin; Li, Licui; Hsiang, Tom; Wang, Maosheng; Shang, Shenghua; Yu, Zhihe

    2016-08-05

    Tobacco grey mold caused by Botrytis cinerea is an important fungal disease worldwide. Boscalid, carbendazim, iprodione, pyrimethanil and propiconazole are representative botryticides for grey mold management. This research investigated the sensitivities of B. cinerea from tobacco to these chemicals using the Biolog FF Microplate. All five chemicals showed inhibitory activity, with average EC50 values of 0.94, 0.05, 0.50, 0.61 and 0.31 μg ml(-1), respectively. B. cinerea metabolized 96.8% of tested carbon sources, including 29 effectively and 33 moderately, but the metabolic fingerprints differed under pressures imposed by these botryticides. For boscalid, B. cinerea was unable to metabolize many substrates related to tricarboxylic acid cycle. For carbendazim, carbon sources related to glycolysis were not metabolized. For iprodione, use of most carbon substrates was weakly inhibited, and the metabolic profile was similar to that of the control. For propiconazole, no carbon substrates were metabolized and the physiological and biochemical functions of the pathogen were totally inhibited. These findings provide useful information on metabolic activities of these botryticides, and may lead to future applications of the Biolog FF Microplate for examining metabolic effects of other fungicides on other fungi, as well as providing a metabolic fingerprint of B. cinerea that could be useful for identification.

  11. Antifungal activity of zinc oxide nanoparticles against Botrytis cinerea and Penicillium expansum.

    Science.gov (United States)

    He, Lili; Liu, Yang; Mustapha, Azlin; Lin, Mengshi

    2011-03-20

    Antifungal activities of zinc oxide nanoparticles (ZnO NPs) and their mode of action against two postharvest pathogenic fungi (Botrytis cinerea and Penicillium expansum) were investigated in this study. ZnO NPs with sizes of 70 ± 15 nm and concentrations of 0, 3, 6 and 12 mmol l(-1) were used. Traditional microbiological plating, scanning electron microscopy (SEM), and Raman spectroscopy were used to study antifungal activities of ZnO NPs and to characterize the changes in morphology and cellular compositions of fungal hyphae treated with ZnO NPs. Results show that ZnO NPs at concentrations greater than 3 mmol l(-1) can significantly inhibit the growth of B. cinerea and P. expansum. P. expansum was more sensitive to the treatment with ZnO NPs than B. cinerea. SEM images and Raman spectra indicate two different antifungal activities of ZnO NPs against B. cinerea and P. expansum. ZnO NPs inhibited the growth of B. cinerea by affecting cellular functions, which caused deformation in fungal hyphae. In comparison, ZnO NPs prevented the development of conidiophores and conidia of P. expansum, which eventually led to the death of fungal hyphae. These results suggest that ZnO NPs could be used as an effective fungicide in agricultural and food safety applications.

  12. Polyamines attenuate ethylene-mediated defense responses to abrogate resistance to Botrytis cinerea in tomato.

    Science.gov (United States)

    Nambeesan, Savithri; AbuQamar, Synan; Laluk, Kristin; Mattoo, Autar K; Mickelbart, Michael V; Ferruzzi, Mario G; Mengiste, Tesfaye; Handa, Avtar K

    2012-02-01

    Transgenic tomato (Solanum lycopersicum) lines overexpressing yeast spermidine synthase (ySpdSyn), an enzyme involved in polyamine (PA) biosynthesis, were developed. These transgenic lines accumulate higher levels of spermidine (Spd) than the wild-type plants and were examined for responses to the fungal necrotrophs Botrytis cinerea and Alternaria solani, bacterial pathogen Pseudomonas syringae pv tomato DC3000, and larvae of the chewing insect tobacco hornworm (Manduca sexta). The Spd-accumulating transgenic tomato lines were more susceptible to B. cinerea than the wild-type plants; however, responses to A. solani, P. syringae, or M. sexta were similar to the wild-type plants. Exogenous application of ethylene precursors, S-adenosyl-Met and 1-aminocyclopropane-1-carboxylic acid, or PA biosynthesis inhibitors reversed the response of the transgenic plants to B. cinerea. The increased susceptibility of the ySpdSyn transgenic tomato to B. cinerea was associated with down-regulation of gene transcripts involved in ethylene biosynthesis and signaling. These data suggest that PA-mediated susceptibility to B. cinerea is linked to interference with the functions of ethylene in plant defense.

  13. Baseline sensitivity to fluopyram and fungicide resistance phenotypes of botrytis cinerea populations from table grapes in california

    Science.gov (United States)

    Gray mold caused by Botrytis cinerea is a major postharvest disease of table grapes grown in the Central Valley of California. Understanding fungicide-resistant phenotypes of B. cinerea is important to the development of pre-harvest fungicide programs for control of postharvest gray mold. Baseline s...

  14. Pectin degradation by Botrytis cinerea: recognition of endopolygalacturonases by an Arabidopsis receptor and utilization of Dgalacturonic acid

    NARCIS (Netherlands)

    Lisha Zhang, Lisha

    2013-01-01

    The necrotrophic fungal plant pathogenBotrytis cinerea is able to infect over 200 host plants and cause severe damage to crops, both pre- and post-harvest. B. cinerea often penetrates host leaf tissue at the anticlinal cell wall and subsequently grows into and through the middle lamella, which consi

  15. The pOT and pLOB vector systems: Improving ease of transgene expression in Botrytis cinerea

    NARCIS (Netherlands)

    Patel, R.M.; Heneghan, M.N.; Kan, van J.A.L.; Bailey, A.M.; Foster, G.D.

    2008-01-01

    This paper outlines the construction of a novel vector system comprising interchangeable terminators, as well as a multiple cloning site (MCS), to facilitate the transformation of the fungal plant pathogen Botrytis cinerea. Previous molecular studies on B. cinerea have relied upon the pLOB1 based ve

  16. Botrytis cinerea Control and the Problem of Fungicide Resistance

    Directory of Open Access Journals (Sweden)

    Brankica Tanović

    2011-01-01

    Full Text Available Botrytis cinerea, the causal agent of grey mould, greatly affects fruit, grapevine, vegetable and ornamental crops production. It is a common causal agent of diseases in plants grown in protected areas, as well as fruit decay during storage and transport. The fungusinvades almost all parts of the plant in all developmental stages, and the symptoms are usually described as grey mould, grey mildew, brown rot and seedling blight. The paper reviews the current knowledge on control possibilities of this necrotrophic pathogen. Theattention is particularly paid to the mode of action of novel fungicides and to the problem of resistance. It is pointed out that by limiting the number of treatments in the growing season, avoiding the use of only one fungicide with a high risk for resistance development,appropriate application rate and timing, using mixtures of pesticides with different modes of action, as well as by alternative use of pesticides from different resistance groups, a longterm preservation of pesticide efficacy is provided.

  17. Novel Coprinopsis cinerea polyesterase that hydrolyzes cutin and suberin.

    Science.gov (United States)

    Kontkanen, Hanna; Westerholm-Parvinen, Ann; Saloheimo, Markku; Bailey, Michael; Rättö, Marjaana; Mattila, Ismo; Mohsina, Marzia; Kalkkinen, Nisse; Nakari-Setälä, Tiina; Buchert, Johanna

    2009-04-01

    Three cutinase gene-like genes from the basidiomycete Coprinopsis cinerea (Coprinus cinereus) found with a similarity search were cloned and expressed in Trichoderma reesei under the control of an inducible cbh1 promoter. The selected transformants of all three polyesterase constructs showed activity with p-nitrophenylbutyrate, used as a model substrate. The most promising transformant of the cutinase CC1G_09668.1 gene construct was cultivated in a laboratory fermentor, with a production yield of 1.4 g liter(-l) purified protein. The expressed cutinase (CcCUT1) was purified to homogeneity by immobilized metal affinity chromatography exploiting a C-terminal His tag. The N terminus of the enzyme was found to be blocked. The molecular mass of the purified enzyme was determined to be around 18.8 kDa by mass spectrometry. CcCUT1 had higher activity on shorter (C(2) to C(10)) fatty acid esters of p-nitrophenol than on longer ones, and it also exhibited lipase activity. CcCUT1 had optimal activity between pH 7 and 8 but retained activity over a wide pH range. The enzyme retained 80% of its activity after 20 h of incubation at 50 degrees C, but residual activity decreased sharply at 60 degrees C. Microscopic analyses and determination of released hydrolysis products showed that the enzyme was able to depolymerize apple cutin and birch outer bark suberin.

  18. The Herbicidal Activity of Mutant Isolates from Botrytis cinerea

    Institute of Scientific and Technical Information of China (English)

    ZHANG Jin-lin; ZHANG Li-hui; LIU Ying-chao; MA Juan; LI Chuan; DONG Jin-gao

    2006-01-01

    Fifteen mutant isolates were obtained by ultraviolet mutation from parent isolate Botrytis cinerea BC-4. Among them three mutant isolates, BC4-1, BC4-2, and BC4-15, showed strong herbicidal activity. BC4-1 showed maximum herbicidal activity for inhibition of germination and growth of Digitaria sanguinalis L. and Amaranthus retroflexus L. The results also showed that herbicidal activity was influenced by differing pH of PD media, with pH value of 4.0 being the optimum.The crude toxin was extracted using chloroform, petroleum ether, and ethyl acetate, respectively, and the ethyl acetate extracts showed the strongest inhibitory activity on the germination and growth of D. sanguinalis L. and A. retroflexus L.Using HPLC, one fraction with an absorption peak at 271 nm was separated from the crude toxin. This fraction could strongly inhibit the growth of D. sanguinalis L. at a concentration of 100 mg L-1 and could completely inhibit the seed germination of D. sanguinalis L. and A. retroflexus L. at a concentration of 50 mg L-1.

  19. Difficulties in differentiating Neisseria cinerea from Neisseria gonorrhoeae in rapid systems used for identifying pathogenic Neisseria species.

    Science.gov (United States)

    Boyce, J M; Mitchell, E B

    1985-11-01

    Neisseria cinerea and Neisseria gonorrhoeae may occur at the same body sites and may have similar colony morphologies. Ideally, systems used for rapid identification of N. gonorrhoeae should be able to differentiate N. cinerea from gonococci. We tested seven N. cinerea strains using the Gonochek II (Du Pont Diagnostics), Minitek (BBL Microbiology Systems), RapID-NH (Innovative Diagnostics, Inc.), RIM-N (American Microscan), and Phadebact (Pharmacia Diagnostics) systems. We found that the reactions produced by N. cinerea in Gonochek II, Minitek, and RapID-NH kits could be confused with the results produced by some strains of N. gonorrhoeae. The susceptibility of N. cinerea to colistin, its ability to grow on tryptic soy or Mueller-Hinton agar, and its inability to grow on modified Thayer-Martin medium help differentiate it from gonococci.

  20. Optimization of Production Xylanase from Marine Bacterium Bacillus safensis P20 on Sugarcane Baggase by Submerged Fermentation

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    Nanik Rahmani

    2014-01-01

    Full Text Available Endo-1, 4-β-xylanase commonly called xylanase is an industrially important enzyme which degrades of lignocellulosic materials to sugar, alcohol and other useful product. The use of commercially xylan is too expensive for use at industrial scale production. For commercial applications, xylanases should ideally be produced from simple and inexpensive substrates. Indonesia has abundantly agro-residues such as sugar cane bagasse which is attractive to be used as carbon sources for the production of enzyme.  In this study, optimization of fermentation condition extracellular xylanasefrom marine bacterium, Bacillus safensisP20 has been conducted by using sugarcane bagasse as carbon source under sub merged fermentation (SMF. Maximum xylanase production was obtained at sugar cane bagasse concentration 1.5%, pH medium 7, and temperature fermentation 20oC, lactose as a carbone source and urea as a nitrogen source with activity 4.06 U/mL for 96 hours.

  1. The study on the infection of apple fruits by Botrytis cinerea Pers. after harvest

    Directory of Open Access Journals (Sweden)

    Henryk Bryk

    2013-12-01

    Full Text Available The aim of this studv was to determine the possibility to infection of apples after harvest by conidia and/or mycelium of Botrytis cinerea Pers. Conidia were unable to infect uninjured apple skin regardless of inoculum density and presence of nutrients. The infection of apples by conidia occurred after the surface wax had been removed by washing of apples with chloroform. Injuries of skin appeared to be a favourable entry point for conidia and mycelium of B.cinerea. Only the mycelium of B.cinerea developed on the apple but not that grown on the artificial medium (PDA was able to directly penetration uninjured apple skin. It was observed that sometimes rotted spots develo ped arround the lenticels.

  2. Transformation of Botrytis cinerea by direct hyphal blasting or by wound-mediated transformation of sclerotia

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    Ish - Shalom Shahar

    2011-12-01

    Full Text Available Abstract Background Botrytis cinerea is a haploid necrotrophic ascomycete which is responsible for 'grey mold' disease in more than 200 plant species. Broad molecular research has been conducted on this pathogen in recent years, resulting in the sequencing of two strains, which has generated a wealth of information toward developing additional tools for molecular transcriptome, proteome and secretome investigations. Nonetheless, transformation protocols have remained a significant bottleneck for this pathogen, hindering functional analysis research in many labs. Results In this study, we tested three different transformation methods for B. cinerea: electroporation, air-pressure-mediated and sclerotium-mediated transformation. We demonstrate successful transformation with three different DNA constructs using both air-pressure- and sclerotium-mediated transformation. Conclusions These transformation methods, which are fast, simple and reproducible, can expedite functional gene analysis of B. cinerea.

  3. Expansive phenotypic landscape of Botrytis cinerea shows differential contribution of genetic diversity and plasticity

    DEFF Research Database (Denmark)

    Corwin, Jason A; Subedy, Anushriya; Eshbaugh, Robert

    2016-01-01

    and genetic diversity for virulence-associated phenotypes in a generalist plant pathogen, we grew a population of 15 isolates of Botrytis cinerea from throughout the world, under a variety of in vitro and in planta conditions. Under in planta conditions, phenotypic differences between the isolates were...... the phenotypic variation under in vitro experimental conditions to phenotypic variation during plant infection. This study indicates that there is a high level of phenotypic variation within B. cinerea that is controlled by a mixture of genetic variation, environment, and genotype × environment. This argues...... that future experiments into the pathogenicity of B. cinerea must account for the genetic and environmental variation within the pathogen to better sample the potential phenotypic space of the pathogen....

  4. Dataset of the Botrytis cinerea phosphoproteome induced by different plant-based elicitors

    Directory of Open Access Journals (Sweden)

    Eva Liñeiro

    2016-06-01

    Full Text Available Phosphorylation is one of the main post-translational modification (PTM involved in signaling network in the ascomycete Botrytis cinerea, one of the most relevant phytopathogenic fungus. The data presented in this article provided a differential mass spectrometry-based analysis of the phosphoproteome of B. cinerea under two different phenotypical conditions induced by the use of two different elicitors: glucose and deproteinized Tomate Cell Walls (TCW. A total 1138 and 733 phosphoproteins were identified for glucose and TCW culture conditions respectively. Raw data are deposited at the ProteomeXchange Consortium via the PRIDE partner repository with the data set identifier (PRIDE: http://www.ebi.ac.uk/pride/archive/projects/PXD003099. Further interpretation and discussion of these data are provided in our research article entitled “Phosphoproteome analysis of B.cinerea in response to different plant-based elicitors” (Liñeiro et al., 2016 [1].

  5. Improvement of xylanase production by Aspergillus niger XY-1 using response surface methodology for optimizing the medium composition

    Institute of Scientific and Technical Information of China (English)

    Yao-xing XU; Yan-li LI; Shao-chun XU; Yong LIU; Xin WANG; Jiang-wu TANG

    2008-01-01

    Objective: To study the optimal medium composition for xylanase production by Aspergillus niger XY-1 in solid-state fermentation (SSF). Methods: Statistical methodology including the Plackett-Burman design (PBD) and the central composite design (CCD) was employed to investigate the individual crucial component of the medium that significantly affected the enzyme yield. Results: Firstly, NaNO3, yeast extract, urea, Na2CO3, MgSO4, peptone and (NH4)2SO4 were screened as the significant factors positively affecting the xylanase production by PBD. Secondly, by valuating the nitrogen sources effect, urea was proved to be the most effective and economic nitrogen source for xylanase production and used for further optimization.Finally, the CCD and response surface methodology (RSM) were applied to determine the optimal concentration of each sig-nificant variable, which included urea, Na2CO3 and MgSO4. Subsequently a second-order polynomial was determined by mul-tiple regression analysis. The optimum values of the critical components for maximum xylanase production were obtained as follows: x1 (urea)=0.163 (41.63 g/L), x2 (Na2CO3)=-1.68 (2.64 g/L), x3 (MGSO4)=1.338 (10.68 g/L) and the predicted xylanase value was 14374.6 U/g dry substrate. Using the optimized condition, xylanase production by Aspergillus niger XY-1 after 48 h fermentation reached 14637 U/g dry substrate with wheat bran in the shake flask. Conclusion: By using PBD and CCD, we obtained the optimal composition for xylanase production by Aspergillus niger XY-1 in SSF, and the results of no additional expensive medium and shortened fermentation time for higher xylanase production show the potential for industrial utilization.

  6. Thermal inactivation of Botrytis cinerea conidia in synthetic medium and strawberry puree.

    Science.gov (United States)

    Villa-Rojas, R; Sosa-Morales, M E; López-Malo, A; Tang, J

    2012-04-16

    Botrytis cinerea is one of the most important post-harvest molds that cause quality deterioration of strawberries and other fruits even during refrigeration storage. This research studied the effects of thermal inactivation of B. cinerea in synthetic medium and strawberry puree using hot water baths at different temperatures. These media were studied in order to determine if results obtained in a solution with the major components of the fruit (synthetic media), are comparable to the ones obtained in fruit purees. The results demonstrated that B. cinerea spores can be inactivated by heat treatments using relatively low temperatures (42-46 °C). Inactivation curves were well described by first order kinetics (R² 0.91-0.99). B. cinerea conidia inoculated in synthetic medium required less time to achieve one log reduction in population than those inoculated in the fruit puree. D values were 22, 8.5, 4 and 1.4 min at 42, 44, 46 and 48 °C, respectively, in synthetic medium; while D values in strawberry puree were 44.9, 13.8, 4.7 and 1.4 min at 42, 44, 46 and 48 °C, respectively. The z values obtained were 4.15 and 5.08 °C for the strawberry puree and synthetic medium respectively, showing higher sensitivity of B. cinerea in fruit purees than in the synthetic medium. Thus, a change in the medium composition had a marked difference in the heat inactivation of B. cinerea conidia, and the results obtained in synthetic medium are not accurate to describe the behavior of the microorganism in the fruit.

  7. Genomic analysis of the necrotrophic fungal pathogens Sclerotinia sclerotiorum and Botrytis cinerea.

    Directory of Open Access Journals (Sweden)

    Joelle Amselem

    2011-08-01

    Full Text Available Sclerotinia sclerotiorum and Botrytis cinerea are closely related necrotrophic plant pathogenic fungi notable for their wide host ranges and environmental persistence. These attributes have made these species models for understanding the complexity of necrotrophic, broad host-range pathogenicity. Despite their similarities, the two species differ in mating behaviour and the ability to produce asexual spores. We have sequenced the genomes of one strain of S. sclerotiorum and two strains of B. cinerea. The comparative analysis of these genomes relative to one another and to other sequenced fungal genomes is provided here. Their 38-39 Mb genomes include 11,860-14,270 predicted genes, which share 83% amino acid identity on average between the two species. We have mapped the S. sclerotiorum assembly to 16 chromosomes and found large-scale co-linearity with the B. cinerea genomes. Seven percent of the S. sclerotiorum genome comprises transposable elements compared to <1% of B. cinerea. The arsenal of genes associated with necrotrophic processes is similar between the species, including genes involved in plant cell wall degradation and oxalic acid production. Analysis of secondary metabolism gene clusters revealed an expansion in number and diversity of B. cinerea-specific secondary metabolites relative to S. sclerotiorum. The potential diversity in secondary metabolism might be involved in adaptation to specific ecological niches. Comparative genome analysis revealed the basis of differing sexual mating compatibility systems between S. sclerotiorum and B. cinerea. The organization of the mating-type loci differs, and their structures provide evidence for the evolution of heterothallism from homothallism. These data shed light on the evolutionary and mechanistic bases of the genetically complex traits of necrotrophic pathogenicity and sexual mating. This resource should facilitate the functional studies designed to better understand what makes these

  8. Research on microorganism xylanases and their applications%微生物木聚糖酶及其应用

    Institute of Scientific and Technical Information of China (English)

    高艳秀; 陈复生; 丁长河

    2012-01-01

    Xylan is the major constituent of hemicelluloses. Due to the structural heterogeneity of the xylans, xylan-degrading enzyme systems include several hydrolytic enzymes. Xylanase(EC 3.2.1.8,1,4-β-D-xylanase)can hydrolyze β-1,4-glycosidic linkages of the xylan backbone to produce short chain xylooligosacchrides of various lengths. Hence, endo-β - xylanase is the crucial enzyme component of microbial xylanolytic enzyme systems. And the microorganism xylanases system, classification, source and distribution were summarized. The microorganism xylanases including their properties, production and their applications in food, papermaking, feed industries were reviewed.%木聚糖(Xylan)是植物半纤维素的主要成分,是一种复杂的多聚五碳糖.木聚糖酶(Xylanase,EC 3.2.1.8)以内切方式作用于木聚糖主链,产生不同链长的寡糖和少量的木糖,是木聚糖降解酶系中最为关键的酶.本文综述了微生物木聚糖酶系统、微生物木聚糖酶的分类及其来源分布、微生物木聚糖酶的特性、产生及在食品、造纸、饲料行业的应用.

  9. Investigating the expression of F10 and G11 xylanases in Aspergillus niger A09 with qPCR.

    Science.gov (United States)

    Cui, Shixiu; Wang, Tianwen; Hu, Hong; Liu, Liangwei; Song, Andong; Chen, Hongge

    2016-09-01

    There exist significant differences between the 2 main types of xylanases, family F10 and G11. A clear understanding of the expression pattern of microbial F10 and G11 under different culture conditions would facilitate better production and industrial application of xylanase. In this study, the fungal xylanase producer Aspergillus niger A09 was systematically investigated in terms of induced expression of xylanase F10 and G11. Results showed that carbon and nitrogen sources could influence xylanase F10 and G11 transcript abundance, with G11 more susceptible to changes in culture media composition. The most favorable carbon and nitrogen sources for high G11 and low F10 production by A. niger A09 were xylan (2%) and (NH4)2C2O4 (0.3%), respectively. Following cultivation at 33 °C for 60 h, the highest xylanase activity (1132 IU per gram of wet mycelia) was observed. On the basis of differential gene expression of F10 and G11, as well as their different properties, we deduced that the F10 protein initially targeted xylan and hydrolyzed it into fragments including xylose, after which xylose acted as the inducer of F10 and G11 gene expression. These speculations also accounted for our failure to identify conditions favoring the high production of F10 but a low production of G11.

  10. Paddy Husk as Support for Solid State Fermentation to Produce Xylanase from Bacillus pumilus

    Institute of Scientific and Technical Information of China (English)

    Ranganathan KAPILAN; Vasanthy ARASARATNAM

    2011-01-01

    To optimize culture conditions for xylanase production by solid state fermentation (SSF) using Bacillus pumilus,with paddy husk as support,solid medium contained 200 g of paddy husk with 800 mL of liquid fermentation medium [xylan,20.0 g/L; peptone,2.0 g/L; yeast extract,2.5 g/L; K2HPO4,2.5 g/L; KH2PO4,1.0 g/L; NaCl,0.1 g/L; (NH4)2SO4,2.0 g/L,CaCl22H2O,0.005 g/L; MgCl2·6H2O,0.005 g/L; and FeCl3,0.005 g/L] at pH 9.0 was applied.The highest xylanase activity (142.0 +0.47 U/g DM] was obtained on the 6th day at 30℃.The optimized paddy husk to liquid fermentation medium ratio was 2∶9,and the optimized culture temperature was 40℃.When commercial Birchwood xylan was replaced with different concentrations of corncob,xylanase production was maximized (224.2 U/g DM) in the medium with 150 g/L corncob.Xylanase production was increased by sucrose,fructose and arabinose,whereas reduced by glucose,galactose,lactose and amylose.When organic nitrogen sources were replaced with locally available nitrogen sources such as groundnut powder or sesame seedcake powder or coconut seedcake powder or soy meal powder,the highest xylanase production (290.7 U/g DM) was obtained in the medium with soy meal powder and 16.0 g/L of soy meal powder was the optimum (326.5±0.34 U/g DM).Based on the optimization studies,B.pumilus produced 2.3 times higher xylanase activity.The medium cost was reduced from 2 458.3 to 178.3 SLR/kg and the total activity which could be obtained from 1 kg of the medium was increased from 48 624 to 220 253 Units.

  11. Production, purification and characterisation of alkali stable xylanase from Cellulosimicrobium sp. MTCC 10645

    Institute of Scientific and Technical Information of China (English)

    Rajashri D Kamble; Anandrao R Jadhav

    2012-01-01

    Objective: The aim of this experimental study was production, purification and characterization of alkali stable xylanase from locally isolated Cellulosimicrobium sp. MTCC 10645, which is an important industrial enzyme used in the pulp and paper industry. Methods: The enzyme was produced in Erlenmeyer flasks containing fresh basal salt medium supplemented with 1% oat spelt xylan. The enzyme was extracted and isolated using ammonium sulphate precipitation and dialysis. It was further purified using DEAE cellulose chromatography and purity was checked by SDS-PAGE. Effect of temperature and pH on activity and stability of enzyme was studied. The enzyme was laso studied for its substrate specificity and kinetic parameters. Results: The isolate was identified on the basis of cultural, morphological, physiological and biochemical properties as well as 16S rRNA sequencing. Among the carbon sources tested, birchwood xylan found prominent for increased level of xylanase i. e. 96.33 U/ml. The enzyme was purified by DEAE cellulose chromatography at NaCl concentration of 0.25 M and had a molecular mass of 78.0 kDa. Xylanase was purified sixteen fold with a specific activity of 246.6 U/mg. Xylanase activity was maximum at 50℃. The enzyme was thermostable retaining 8%of the original activity after incubation at 60℃ of 4 h. The enzyme was active over a pH range of 6.0-11.0, although its activity was optimal at pH 7.0. About 48.52% of the enzyme activity was retained after 4 h at pH 11.0. The enzyme was active on oat spelt and birchwood xylans but not on avicel, CMC, cellobiose, starch or p-nitrophenyl xylopyranoside. The xylanase had Km and Vmax values of 4.76 mg/ml and 232.5 μmol/min/mg, respectively when birchwood xylan used as substrate. Conclusions:The xylanase showed a unique pattern of xylan hydrolysis releasing a large amount of intermediate products (xylotriose and xylobiose) with small quantity of xylose. Some of these characteristics make this enzyme potentially

  12. The study on the infection of apple fruits by Botrytis cinerea Pers. after harvest

    OpenAIRE

    Henryk Bryk

    2013-01-01

    The aim of this studv was to determine the possibility to infection of apples after harvest by conidia and/or mycelium of Botrytis cinerea Pers. Conidia were unable to infect uninjured apple skin regardless of inoculum density and presence of nutrients. The infection of apples by conidia occurred after the surface wax had been removed by washing of apples with chloroform. Injuries of skin appeared to be a favourable entry point for conidia and mycelium of B.cinerea. Only the mycelium of B.cin...

  13. Characterization of Botrytis cinerea isolates from small fruits and grapevine in Serbia

    Directory of Open Access Journals (Sweden)

    Tanović Brankica

    2009-01-01

    Full Text Available Twenty-six single-spore isolates of Botrytis cinerea from blackberry, raspberry, strawberry, and grapevine were investigated using transposable elements, morphological characterization, and sensitivity to fungicides. Both transposable elements, Flipper and Boty, were detected among isolates from all the hosts. Six vacuma (without transposable elements and seven transposa (containing both elements isolates were found to be present in sympatry in Serbia. Isolates containing only the Boty element were detected. Eight morphological types of colonies on PDA and MA media were observed, confirming the great phenotypic variability of B. cinerea. Sensitivity to fungicides was various, depending on both the fungicide and the isolate.

  14. Filamentous fungi isolated from grape marc as antagonists of Botrytis cinerea

    Directory of Open Access Journals (Sweden)

    Jovičić-Petrović Jelena P.

    2016-01-01

    Full Text Available In this paper we report on the isolation and identification of three filamentous fungi from grape marc, and antifungal effect of their cell-free culture filtrates on the growth of Botrytis cinerea, causal agent of gray mold. Grape marc is a waste material that has been used as soil amendment in sustainable agriculture. Isolates originating from grape marc were identified on the basis of morphological features and internal transcribed spacer rDNA or β-tubulin gene sequencing. The presence of three different species, Penicillium paneum, Penicillium chrysogenum and Aspergillus fumigatus has been detected expressing different effect on the growth of B. cinerea. The effect of crude culture filtrates of selected fungi on B. cinerea growth was tested. Heat sensitivity of the established inhibition effect was examined by autoclaving the crude culture filtrate prior to testing. Additional aim was to determine whether antifungal effect was influenced by previous exposure to B. cinerea in dual liquid cultures. Crude culture filtrate of A. fumigatus K16/2 showed the lowest suppression of B. cinerea growth. A maximal percentage inhibition achieved within the study was 38.2%, 39.8% and 23.8 for crude filtrates of P. paneum K7/1, P. chrysogenum K11/1 and A. fumigatus K16/2, respectively. Presence of B. cinerea in dual liquid culture induced significant increase in antifungal capacity of the culture filtrates in comparison to pure culture filtrates of the chosen isolates. The antifungal activity of all of the isolates’ culture filtrates retained after heat treatment suggesting the presence of some thermostable antifungal metabolites. The results indicate the complexity and specificity of the interaction between filamentous fungi and B. cinerea. Grape marc is a good source for isolation od B. cinerea fungal antagonists and their antifungal metabolites. Specificity of fungal-fungal interactions suggests that further research on the antagonistic mechanisms and

  15. Cloning of xylanase gene of Streptomyces flavogriseus in Escherichia coli and bacteriophage lambda-induced lysis for the release of cloned enzyme.

    Science.gov (United States)

    Srivastava, R; Ali, S S; Srivastava, B S

    1991-03-01

    The xylanase gene of Streptomyces flavogriseus was cloned in pUC8 plasmid and expressed in Escherichia coli lysogenic for lambda cI857. lambda-Induced lysis of E. coli at 42 degrees C allowed efficient release of cloned enzyme activity in extracellular environment. The xylanase gene was located in the 0.8-kb HindIII fragment and coded for 18,000 Mr xylanase.

  16. The expression of a xylanase targeted to ER-protein bodies provides a simple strategy to produce active insoluble enzyme polymers in tobacco plants.

    Directory of Open Access Journals (Sweden)

    Immaculada Llop-Tous

    Full Text Available BACKGROUND: Xylanases deserve particular attention due to their potential application in the feed, pulp bleaching and paper industries. We have developed here an efficient system for the production of an active xylanase in tobacco plants fused to a proline-rich domain (Zera of the maize storage protein γ-zein. Zera is a self-assembling domain able to form protein aggregates in vivo packed in newly formed endoplasmic reticulum-derived organelles known as protein bodies (PBs. METHODOLOGY/PRINCIPAL FINDINGS: Tobacco leaves were transiently transformed with a binary vector containing the Zera-xylanase coding region, which was optimized for plant expression, under the control of the 35S CaMV promoter. The fusion protein was efficiently expressed and stored in dense PBs, resulting in yields of up to 9% of total protein. Zera-xylanase was post-translationally modified with high-mannose-type glycans. Xylanase fused to Zera was biologically active not only when solubilized from PBs but also in its insoluble form. The resistance of insoluble Zera-xylanase to trypsin digestion demonstrated that the correct folding of xylanase in PBs was not impaired by Zera oligomerization. The activity of insoluble Zera-xylanase was enhanced when substrate accessibility was facilitated by physical treatments such as ultrasound. Moreover, we found that the thermostability of the enzyme was improved when Zera was fused to the C-terminus of xylanase. CONCLUSION/SIGNIFICANCE: In the present work we have successfully produced an active insoluble aggregate of xylanase fused to Zera in plants. Zera-xylanase chimeric protein accumulates within ER-derived protein bodies as active aggregates that can easily be recovered by a simple density-based downstream process. The production of insoluble active Zera-xylanase protein in tobacco outlines the potential of Zera as a fusion partner for producing enzymes of biotechnological relevance. Zera-PBs could thus become efficient and low

  17. Emerging role of N- and C-terminal interactions in stabilizing (β/α8 fold with special emphasis on Family 10 xylanases

    Directory of Open Access Journals (Sweden)

    Amit Bhardwaj

    2012-09-01

    Full Text Available Xylanases belong to an important class of industrial enzymes. Various xylanases have been purified and characterized from a plethora of organisms including bacteria, marine algae, plants, protozoans, insects, snails and crustaceans. Depending on the source, the enzymatic activity of xylanases varies considerably under various physico-chemical conditions such as temperature, pH, high salt and in the presence of proteases. Family 10 or glycosyl hydrolase 10 (GH10 xylanases are one of the well characterized and thoroughly studied classes of industrial enzymes. The TIM-barrel fold structure which is ubiquitous in nature is one of the characteristics of family 10 xylanases. Family 10 xylanases have been used as a “model system” due to their TIM-barrel fold to dissect and understand protein stability under various conditions. A better understanding of structure-stability-function relationships of family 10 xylanases allows one to apply these governing molecular rules to engineer other TIM-barrel fold proteins to improve their stability and retain function(s under adverse conditions. In this review, we discuss the implications of N-and C-terminal interactions, observed in family 10 xylanases on protein stability under extreme conditions. The role of metal binding and aromatic clusters in protein stability is also discussed. Studying and understanding family 10 xylanase structure and function, can contribute to our protein engineering knowledge.

  18. EMERGING ROLE OF N- AND C-TERMINAL INTERACTIONS IN STABILIZING (β;/α8 FOLD WITH SPECIAL EMPHASIS ON FAMILY 10 XYLANASES

    Directory of Open Access Journals (Sweden)

    Amit Bhardwaj

    2012-09-01

    Full Text Available Xylanases belong to an important class of industrial enzymes. Various xylanases have been purified and characterized from a plethora of organisms including bacteria, marine algae, plants, protozoans, insects, snails and crustaceans. Depending on the source, the enzymatic activity of xylanases varies considerably under various physico-chemical conditions such as temperature, pH, high salt and in the presence of proteases. Family 10 or glycosyl hydrolase 10 (GH10 xylanases are one of the well characterized and thoroughly studied classes of industrial enzymes. The TIM-barrel fold structure which is ubiquitous in nature is one of the characteristics of family 10 xylanases. Family 10 xylanases have been used as a “model system” due to their TIM-barrel fold to dissect and understand protein stability under various conditions. A better understanding of structure-stability-function relationships of family 10 xylanases allows one to apply these governing molecular rules to engineer other TIM-barrel fold proteins to improve their stability and retain function(s under adverse conditions. In this review, we discuss the implications of N-and C-terminal interactions, observed in family 10 xylanases on protein stability under extreme conditions. The role of metal binding and aromatic clusters in protein stability is also discussed. Studying and understanding family 10 xylanase structure and function, can contribute to our protein engineering knowledge.

  19. Toxicological studies on Thermomyces lanuginosus xylanase expressed by Fusarium venenatum, intended for use in food.

    Science.gov (United States)

    Pedersen, P B; Broadmeadow, A

    2000-09-01

    The xylanase used in this study was produced by a submerged fermentation of Fusarium venenatum and contained a gene code originating from Thermomyces lanuginosus. The enzyme was subject to a 13-week toxicological test in rats and in vitro tests to document its safety in use. The enzyme is to be applied as a processing aid in the baking industry to improve handling and stability of dough. The enzyme was not found to be mutagenic in the Salmonella typhimurium reverse mutation assay, nor did it cause chromosomal aberrations in cultured human lymphocytes. Oral administration to rats of up to 10.0 ml/kg bw/day (equivalent to a Total Organic Solids dosage of 1.12 g/kg bw/day or a xylanase dosage of 89422 FXU (W)/kg bw/day) for 13 weeks did not cause any adverse effect.

  20. Pseudomonas sp. xylanase for clarification of Mausambi and Orange fruit juice

    Science.gov (United States)

    Sharma, Pawan Kumar; Chand, Duni

    2012-07-01

    Xylanase can be usd for many Industrial applications and juice clarification is one of them. Pseudomonas sp. xylanase was used for fruit juice clarification in free State. Maximum amount of juice clarification was in case of Mausambi juice was observed at 40 C∞ and 52 hours, in case of free enzyme treated juice there is 46.9% increase in clarity and 1.7 fold increase in reducing sugars of the juice and enzyme dose was optimized as 8U with maximum flow rate of 6 ml/min at this dose. In case of orange juice in free enzyme treated juice maximum clarity was observed at 40 C∞ and 52 hours, juice was found to be 42.14 % clear with increase of 1.9 fold of reducing sugars, enzyme dose optimized was 8.06U with maximum flow rate of 0.86 ml/min.

  1. Cloning and expression of an endo-1,4-β-xylanase from the coffee berry borer, Hypothenemus hampei

    Directory of Open Access Journals (Sweden)

    Padilla-Hurtado Beatriz

    2012-01-01

    Full Text Available Abstract Background The coffee berry borer, Hypothenemus hampei, reproduces and feeds exclusively on the mature endosperm of the coffee seed, which has a cell wall composed mainly of a heterogeneous mixture of hemicellulose polysaccharides, including arabinoxylans. Xylanases are digestive enzymes responsible for the degradation of xylan based polymers, hydrolyzing them into smaller molecules that are easier to assimilate by insects. We report the cloning, expression and enzymatic characterization of a xylanase gene that was identified in the digestive tract of the coffee berry borer. Methods The complete DNA sequence encoding a H. hampei xylanase (HhXyl was obtained using a genome walking technique in a cDNA library derived from the borer digestive tract. The XIP-I gene was amplified from wheat (Triticum aestivum variety Soisson. A Pichia pastoris expression system was used to express the recombinant form of these enzymes. The xylanase activity and XIP-I inhibitory activity was quantified by the 3,5-dinitrosalicylic (DNS. The biological effects of XIP-I on borer individuals were evaluated by providing an artificial diet enriched with the recombinant XIP-I protein to the insects. Results The borer xylanase sequence contains a 951 bp open reading frame that is predicted to encode a 317-amino acid protein, with an estimated molecular weight of 34.92 kDa and a pI of 4.84. Bioinformatic analysis revealed that HhXyl exhibits high sequence homology with endo-β-D-xylanases of Streptomyces bingchenggensis from glycosyl hydrolase 10 (GH10. The recombinant xylanase showed maximal activity at pH 5.5 and 37°C. XIP-I expressed as a recombinant protein inhibited HhXyl activity in vitro and caused individual H. hampei mortality in bioassays when included as a supplement in artificial diets. Conclusion A xylanase from the digestive tract of the coffee berry borer was identified and functionally characterized. A xylanase inhibitor protein, XIP-I, from wheat was

  2. ABA suppresses Botrytis cinerea elicited NO production in tomato to influence H2O2 generation and increase host susceptibility

    Directory of Open Access Journals (Sweden)

    Anushen eSivakumaran

    2016-05-01

    Full Text Available Abscisic acid (ABA production has emerged a susceptibility factor in plant-pathogen interactions. This work examined the interaction of ABA with NO in tomato following challenge with the ABA-synthesising pathogen, Botrytis cinerea. Trace gas detection using a quantum cascade laser detected NO production within minutes of challenge with B. cinerea whilst photoacoustic laser detection detected ethylene production – an established mediator of defence against this pathogen - occurring after 6 h. Application of the NO generation inhibitor N-Nitro-L-arginine methyl ester (L-NAME suppressed both NO and ethylene production and resistance against B. cinerea. The tomato mutant sitiens fails to accumulate ABA (abscisic acid, shows increased resistance to B. cinerea and we noted exhibited elevated NO and ethylene production. Exogenous application of L-NAME or ABA reduced NO production in sitiens and reduced resistance to B. cinerea. Increased resistance to B. cinerea in sitiens have previously been linked to increased reactive oxygen species (ROS generation but this was reduced in both L-NAME and ABA treated sitiens. Taken together, our data suggests that ABA can decreases resistance to B. cinerea via reduction of NO production which also suppresses both ROS and ethylene production.

  3. LongSAGE gene-expression profiling of Botrytis cinerea germination suppressed by resveratrol, the major grapevine phytoalexin.

    Science.gov (United States)

    Zheng, Chuanlin; Choquer, Mathias; Zhang, Bing; Ge, Hui; Hu, Songnian; Ma, Huiqin; Chen, Shangwu

    2011-09-01

    The ascomycetes Botrytis cinerea is one of the most studied necrotrophic phytopathogens and one of the main fungal parasites of grapevine. As a defense mechanism, grapevine produces a phytoalexin compound, resveratrol, which inhibits germination of the fungal conidium before it can penetrate the plant barriers and lead to host cell necrotrophy. To elucidate the effect of resveratrol on transcriptional regulation in B. cinerea germlings, two LongSAGE (long serial analysis of gene expression) libraries were generated in vitro for gene-expression profiling: 41 428 tags and among them, 15 665 unitags were obtained from resveratrol-treated B. cinerea germlings and 41 358 tags, among them, 16 362 unitags were obtained from non-treated B. cinerea germlings. In-silico analysis showed that about half of these unitags match known genes in the complete B. cinerea genome sequence. Comparison of unitag frequencies between libraries highlighted 110 genes that were transcriptionally regulated in the presence of resveratrol: 53 and 57 genes were significantly down- and upregulated, respectively. Manual curation of their putative functional categories showed that primary metabolism of germinating conidia appears to be markedly affected under resveratrol treatment, along with changes in other putative metabolic pathways, such as resveratrol detoxification and virulence-effector secretion, in B. cinerea germlings. We propose a hypothetical model of cross talk between B. cinerea germinating conidia and resveratrol-producing grapevine at the very early steps of infection.

  4. Effects and possible mechanism of tea tree oil against Botrytis cinerea and Penicillium expansum in vitro and in vivo test.

    Science.gov (United States)

    Li, Yonghua; Shao, Xingfeng; Xu, Jiayu; Wei, Yingying; Xu, Feng; Wang, Hongfei

    2017-03-01

    The purpose of this study was to investigate the antifungal activities and possible mechanisms of tea tree oil (TTO) against Botrytis cinerea and Penicillium expansum in vitro and in vivo. The results show that TTO exhibits dose-dependent antifungal activity against both pathogens, but P. expansum is less sensitive than B. cinerea to TTO not only in the in vitro test but also in artificially inoculated cherry fruits. TTO vapor treatment reduced the decay caused by these pathogens in inoculated cherry fruits, but the effect on P. expansum was less than that on B. cinerea. While the total lipid and ergosterol contents of the cell membrane are greater in P. expansum than in B. cinerea, TTO treatment lowers the total lipid content in the membranes of both species by well over 50%, and ergosterol content is reduced to a greater extent in B. cinerea than in P. expansum. In both pathogens, TTO alters mycelial morphology and cellular ultrastructure. Oxygen consumption measurements show that TTO inhibits respiratory metabolism via the tricarboxylic acid cycle pathway in both pathogens, though more severely in B. cinerea than in P. expansum. The relatively decreased sensitivity of P. expansum to TTO may be due to the fact that TTO causes less disruption of the cell membrane in this organism, and higher inhibition the respiratory metabolism to the extent observed in B. cinerea.

  5. New Host Plant of Botrytis cinerea in China%我国灰葡萄孢(Botrytis cinerea)的新寄主植物

    Institute of Scientific and Technical Information of China (English)

    张中义; 何永宏; 王学英; 张立新; 张陶; 桂明英

    2002-01-01

    @@ 报道我国灰葡萄孢(Botrytis cinerea Pers.: Fr.) 的寄生性新寄主植物73种,按寄主汉名、学名、分布地和标本号排列,标本保藏于云南农业大学真菌标本室(MHYAU),以供植保、植检部门参考.

  6. Effects of disruption of xylanase-encoding genes on the xylanolytic system of Streptomyces lividans.

    Science.gov (United States)

    Arhin, F F; Shareck, F; Kluepfel, D; Morosoli, R

    1994-01-01

    Wild-type Streptomyces lividans produced the three xylanases (XlnA, XlnB, and XlnC) when xylan, xylan hydrolysates obtained by the action of XlnA, XlnB, and XlnC, or purified small xylo-oligosaccharides (xylobiose [X2], xylotriose [X3], xylotetraose [X4], and xylopentaose [X5]) were used as the carbon source. The three xylanase genes of S. lividans (xlnA, xlnB, and xlnC) were disrupted by using vectors that integrate into the respective genes. Disruption of one or more of the xln genes resulted in reduced growth rates and reduced total xylanase activities when the strain was grown in xylan. The greatest effect was observed when xlnA was disrupted. In medium containing xylan, disruption of xlnA did not affect expression of xlnB and xlnC; disruption of xlnB did not affect expression of xlnA but affected expression of xlnC; and disruption of xlnC did not affect expression of xlnA but affected expression of xlnB. A fraction of XlnB or XlnC hydrolytic products (those with a degree of polymerization greater than 11 [X11]) was found to stimulate expression of xlnB and xlnC in strains disrupted in xlnC and xlnB, respectively, whereas lower-molecular-weight fractions as well as purified small xylo-oligosaccharides did not. The stimulating molecule(s) lost its effect when it was hydrolyzed further by XlnA. A mechanism of transglycosylation reactions by the S. lividans xylanases is postulated to be involved in the regulation of xln genes. Images PMID:8051006

  7. Cloning, overexpression, and characterization of a novel alkali-thermostable xylanase from Geobacillus sp. WBI.

    Science.gov (United States)

    Mitra, Suranjita; Mukhopadhyay, Bidhan Chandra; Mandal, Anisur Rahaman; Arukha, Ananta Prasad; Chakrabarty, Kuheli; Das, Gourab Kanti; Chakrabartty, Pran Krishna; Biswas, Swadesh Ranjan

    2015-04-01

    An endo-β-1,4-xylanase gene xynA of a thermophilic Geobacillus sp. WBI from "hot" compost was isolated by PCR amplification. The gene encoding 407 residues were overexpressed in E. coli and purified by Ni-NTA chromatography. The purified enzyme (47 kDa) had a broad pH optimum of 6.0 to 9.0, and was active between 50 and 90 °C. The enzyme retained 100% of its activity when incubated at 65 °C for 1 h under alkaline condition (pH 10.0) and retained 75% activity at pH 11.0. The K(m) and V(max) of the enzyme were 0.9 mg ml(-1) and 0.8 µmol ml(-1) min(-1), respectively. In molecular dynamics simulation at 338 K (65 °C), the enzyme was found to be stable. At an elevated temperature (450 K) specific α-helix and β-turns of the proteins were most denatured. The denaturation was less in WBI compared with its highest homolog G. stearothermophilus T-6 xylanase with difference of six residues. The results predict that these regions are responsible for the improved thermostability observed over related enzymes. The present work encourages further experimental demonstration to understand how these regions contribute thermostability to WBI xylanase. The study noted that WBI produces a xylanase with unique characteristics, specifically alkali-thermostability.

  8. TLXI, a novel type of xylanase inhibitor from wheat (Triticum aestivum) belonging to the thaumatin family.

    Science.gov (United States)

    Fierens, Ellen; Rombouts, Sigrid; Gebruers, Kurt; Goesaert, Hans; Brijs, Kristof; Beaugrand, Johnny; Volckaert, Guido; Van Campenhout, Steven; Proost, Paul; Courtin, Christophe M; Delcour, Jan A

    2007-05-01

    Wheat (Triticum aestivum) contains a previously unknown type of xylanase (EC 3.2.1.8) inhibitor, which is described in the present paper for the first time. Based on its >60% similarity to TLPs (thaumatin-like proteins) and the fact that it contains the Prosite PS00316 thaumatin family signature, it is referred to as TLXI (thaumatin-like xylanase inhibitor). TLXI is a basic (pI> or =9.3 in isoelectric focusing) protein with a molecular mass of approx. 18-kDa (determined by SDS/PAGE) and it occurs in wheat with varying extents of glycosylation. The TLXI gene sequence encodes a 26-amino-acid signal sequence followed by a 151-amino-acid mature protein with a calculated molecular mass of 15.6-kDa and pI of 8.38. The mature TLXI protein was expressed successfully in Pichia pastoris, resulting in a 21-kDa (determined by SDS/PAGE) recombinant protein (rTLXI). Polyclonal antibodies raised against TLXI purified from wheat react with epitopes of rTLXI as well as with those of thaumatin, demonstrating high structural similarity between these three proteins. TLXI has a unique inhibition specificity. It is a non-competitive inhibitor of a number of glycoside hydrolase family 11 xylanases, but it is inactive towards glycoside hydrolase family 10 xylanases. Progress curves show that TLXI is a slow tight-binding inhibitor, with a K(i) of approx. 60-nM. Except for zeamatin, an alpha-amylase/trypsin inhibitor from maize (Zea mays), no other enzyme inhibitor is currently known among the TLPs. TLXI thus represents a novel type of inhibitor within this group of proteins.

  9. Computational design of an endo-1,4-[beta]-xylanase ligand binding site

    Energy Technology Data Exchange (ETDEWEB)

    Morin, Andrew; Kaufmann, Kristian W.; Fortenberry, Carie; Harp, Joel M.; Mizoue, Laura S.; Meiler, Jens (Vanderbilt)

    2012-09-05

    The field of computational protein design has experienced important recent success. However, the de novo computational design of high-affinity protein-ligand interfaces is still largely an open challenge. Using the Rosetta program, we attempted the in silico design of a high-affinity protein interface to a small peptide ligand. We chose the thermophilic endo-1,4-{beta}-xylanase from Nonomuraea flexuosa as the protein scaffold on which to perform our designs. Over the course of the study, 12 proteins derived from this scaffold were produced and assayed for binding to the target ligand. Unfortunately, none of the designed proteins displayed evidence of high-affinity binding. Structural characterization of four designed proteins revealed that although the predicted structure of the protein model was highly accurate, this structural accuracy did not translate into accurate prediction of binding affinity. Crystallographic analyses indicate that the lack of binding affinity is possibly due to unaccounted for protein dynamics in the 'thumb' region of our design scaffold intrinsic to the family 11 {beta}-xylanase fold. Further computational analysis revealed two specific, single amino acid substitutions responsible for an observed change in backbone conformation, and decreased dynamic stability of the catalytic cleft. These findings offer new insight into the dynamic and structural determinants of the {beta}-xylanase proteins.

  10. Two New Xylanases with Different Substrate Specificities from the Human Gut Bacterium Bacteroides intestinalis DSM 17393

    KAUST Repository

    Hong, Pei-Ying

    2014-01-24

    Xylan is an abundant plant cell wall polysaccharide and is a dominant component of dietary fiber. Bacteria in the distal human gastrointestinal tract produce xylanase enzymes to initiate the degradation of this complex heteropolymer. These xylanases typically derive from glycoside hydrolase (GH) families 10 and 11; however, analysis of the genome sequence of the xylan-degrading human gut bacterium Bacteroides intestinalis DSM 17393 revealed the presence of two putative GH8 xylanases. In the current study, we demonstrate that the two genes encode enzymes that differ in activity. The xyn8A gene encodes an endoxylanase (Xyn8A), and rex8A encodes a reducing-end xylose-releasing exo-oligoxylanase (Rex8A). Xyn8A hydrolyzed both xylopentaose (X5) and xylohexaose (X6) to a mixture of xylobiose (X2) and xylotriose (X3), while Rex8A hydrolyzed X3 through X6 to a mixture of xylose (X1) and X2. Moreover, rex8A is located downstream of a GH3 gene (xyl3A) that was demonstrated to exhibit β-xylosidase activity and would be able to further hydrolyze X2 to X1. Mutational analyses of putative active site residues of both Xyn8A and Rex8A confirm their importance in catalysis by these enzymes. Recent genome sequences of gut bacteria reveal an increase in GH8 Rex enzymes, especially among the Bacteroidetes, indicating that these genes contribute to xylan utilization in the human gut.

  11. Gene cloning and characterization of an acidic xylanase from Acidobacterium capsulatum.

    Science.gov (United States)

    Inagaki, K; Nakahira, K; Mukai, K; Tamura, T; Tanaka, H

    1998-06-01

    The gene xynA encoding an acid endo-beta-1,4-xylanase from an acidophilic bacterium, Acidobacterium capsulatum 161, was cloned and expressed in Eschrichia coli. The nucleotide sequence of the 1.6-kb DNA fragment containing xynA was analyzed, revealing an open reading frame of 1,215 bp encoding a peptide of 405 amino acid residues. The deduced amino acid sequence of XynA was very similar to other xylanases that are from the glycosyl hydrolase family 10. XynA was purified to homogeneity by SDS-polyacrylamide gel electrophoresis from E. coli transformants. The molecular mass and isoelectric point of XynA were 41 kDa and 7.3, respectively. The xylanase activity of the cloned XynA is an endo-acting enzyme that shows optimal activity at pH 5.0 and 65 degrees C, and is stable pH between 3.0 and 8.0. The K(m) and Vmax with oat spelt xylan as a substrate at pH 5.0 and 30 degrees C are 3.5 mg/ml and 403 mumol/min/mg.

  12. Optimisation of amylase and xylanase addition in dependance of white flour amylase activity

    Directory of Open Access Journals (Sweden)

    Lončar Davor M.

    2016-01-01

    Full Text Available In this study the effect of different quantities of added amylase to white wheat flours characterized with different activities of naturally existing amylases is tested. Response surface methodology is chosen to test the effects of main applied technological parameters on bread quality responses. Independent variables are chosen to be: quantity of added amylase and bulk fermentation time, while analysed responses are: specific volume, grain structure, bulk fermentation. Bread quality responses are statistically significant, while predicted and observed responses correspond very well, which allows good prediction of bread quality parameters based on applied technological parameters and flour characteristics. Score analysis shows that optimum quantity of amylase addition regarding bread quality depends on the activity of naturally existing amylases. Optimal quantity of added xylanase in bread samples made from both flour types is 0.004%. Xylanase improved properties of white wheat bread and higher effect is experienced with flour that has more active naturally existing amylases. Addition of amylase has statistically significantly increased a* values of crust. Addition of xylanase has statistically significantly decreased values of b* in comparison to the respective bread sample with only added amylase.

  13. Screening of a xylanase clone from a fosmid library of rumen microbiota in Hu sheep.

    Science.gov (United States)

    Wang, Jiakun; Sun, Zhongyuan; Zhou, Yan; Wang, Qian; Ye, Jun'an; Chen, Zhenming; Liu, Jianxin

    2012-01-01

    The glycosyl hydrolase family 11, which is responsible for carbohydrate metabolism, was identified in the open reading frame (ORF) 6 of a xylanase positive clone from a fosmid library of rumen microbiota of Hu sheep. A BLASTP search of GenBank revealed that ORF6 encoded a 355-amino acid putative endoxylanase, having 61% similarity (e(-73)) to endo-1,4-β-xylanase of Fibrobacter succinogenes S85 (YP_003250510.1). Predicted with the SWISS-MODEL, there were two separate β-sandwich clusters linked with a high serine containing linker in ORF6. The N-terminal β-sandwich is a novel endoxylanase of the glycosyl hydrolase family 11 with a specific activity of 1150.00 U/mg. The optimal pH and temperature for this enzyme were shown to be pH 5.0 and 50°C, respectively. The C-terminal helped increase the stability of the xylanase but decreased the activity to some degree. The C-terminal β-sandwich could bind avicel, but no conserved domain could be found. It may be a novel carbohydrate-binding module.

  14. The influence of sorbitol on the production of cellulases and xylanases in an airlift bioreactor.

    Science.gov (United States)

    Ritter, Carla Eliana Todero; Fontana, Roselei Claudete; Camassola, Marli; da Silveira, Maurício Moura; Dillon, Aldo José Pinheiro

    2013-11-01

    The production of cellulases and xylanases by Penicillium echinulatum in an airlift bioreactor was evaluated. In batch production, we tested media with isolated or associated cellulose and sorbitol. In fed-batch production, we tested cellulose addition at two different times, 30 h and 48 h. Higher liquid circulation velocities in the downcomer were observed in sorbitol 10 g L(-1) medium. In batch production, higher FPA (filter paper activity) and endoglucanase activities were obtained with cellulose (7.5 g L(-1)) and sorbitol (2.5 g L(-1)), 1.0 U mL(-1) (120 h) and 6.4 U m L(-1) (100 h), respectively. For xylanases, the best production condition was cellulose 10 g L(-1), which achieved 5.5 U mL(-1) in 64 h. The fed-batch process was favorable for obtaining xylanases, but not for FPA and endoglucanases, suggesting that in the case of cellulases, the inducer must be added early in the process.

  15. Partial purification and characterization of Xylanase from Trichoderma viride produced under SSF

    Directory of Open Access Journals (Sweden)

    M Irfan

    2012-03-01

    Full Text Available Summary: In the present study xylanase enzyme was produced from Trichoderma viride in solid state fermentation using sugarcane bagasse as a substrate. The whole fermentation process was carried out in 250ml Erlenmeyer flask at 30oC for seven days of fermentation period. The enzyme was partially purified by ammonium sulphate (60% fractionation followed by dialysis. The partially purified enzyme was further characterized showing optimum pH and temperature of 5.0 and 50oC respectively. Metal profile of the enzyme showed that it was stimulated by FeSO4 (134%, CaCl2 (129%, BaCl2 (105%, MgSO4 (113%, MnCl2 (102% or AgCl (107% and it was strongly inhibited by EDTA (26% or HgSO4 (32%. Industrial Relevance: In the present study, xylanase enzyme was produced and characterized from Trichoderma viride in solid state fermentation using cheap substrate. This enzyme is very helpful in industrial sector especially in pulp and paper industry, food industry and also in bioethanol production. Pilot scale production of this enzyme in industries can reduce the import cost of the enzyme and make the whole process cost effective. Keywords: Partial purification; Characterization; Xylanase; Trichoderma viride; SSF

  16. Effect of Aspergillus niger xylanase on dough characteristics and bread quality attributes.

    Science.gov (United States)

    Ahmad, Zulfiqar; Butt, Masood Sadiq; Ahmed, Anwaar; Riaz, Muhammad; Sabir, Syed Mubashar; Farooq, Umar; Rehman, Fazal Ur

    2014-10-01

    The present study was conducted to investigate the impact of various treatments of xylanase produced by Aspergillus niger applied in bread making processes like during tempering of wheat kernels and dough mixing on the dough quality characteristics i.e. dryness, stiffness, elasticity, extensibility, coherency and bread quality parameters i.e. volume, specific volume, density, moisture retention and sensory attributes. Different doses (200, 400, 600, 800 and 1,000 IU) of purified enzyme were applied to 1 kg of wheat grains during tempering and 1 kg of flour (straight grade flour) during mixing of dough in parallel. The samples of wheat kernels were agitated at different intervals for uniformity in tempering. After milling and dough making of both types of flour (having enzyme treatment during tempering and flour mixing) showed improved dough characteristics but the improvement was more prominent in the samples receiving enzyme treatment during tempering. Moreover, xylanase decreased dryness and stiffness of the dough whereas, resulted in increased elasticity, extensibility and coherency and increase in volume & decrease in bread density. Xylanase treatments also resulted in higher moisture retention and improvement of sensory attributes of bread. From the results, it is concluded that dough characteristics and bread quality improved significantly in response to enzyme treatments during tempering as compared to application during mixing.

  17. Xylanase production by a thermo-tolerant Bacillus species under solid-state and submerged fermentation

    Directory of Open Access Journals (Sweden)

    Uma Gupta

    2009-12-01

    Full Text Available Effects of xylose on xylanase production by a thermophilic Bacillus sp showed diverse patterns on corn cob (CC and wheat bran (WB as sole carbon sources in solid- state fermentation (SSF and submerged fermentation (SmF. Supplementation of these media with either mineral salt solution (MSS or yeast extract peptone (YEP also exerted variable effects. While under SSF, xylose stimulated xylanase synthesis by 44.01%, on wheat bran supplemented with MSS, it decreased the enzyme activity by 12.89% with YEP supplementation. In SmF, however the enzyme synthesis was stimulated by xylose on supplementation with both MSS and YEP by 41.38% and 27.47%, respectively. On corn cob under SSF, xylose repression was significant both with MSS (26.92% and YEP (23.90% supplementation. Repression by xylose also took place on corn cob and YEP (19.69% under SmF, while significant stimulation (28.55% was observed by MSS supplementation. The possible role of media composition and fermentation conditions in the regulation of xylanase synthesis by xylose is discussed.

  18. Changes in volatile production during an infection of tomato plants by Botrytis cinerea

    NARCIS (Netherlands)

    Jansen, R.M.C.; Miebach, M.; Kleist, E.; Henten, van E.J.; Wildt, J.

    2006-01-01

    Botrytis blight caused by the fungus Botrytis cinerea is probably the most common disease of greenhouse-grown crops like tomato. Botrytis blight in tomato plants is mainly detected by visual inspection or destructive biochemical and molecular determinations. These methods are time consuming and not

  19. Constitutive expression of MKS1 confers susceptibility to Botrytis cinerea infection independent of PAD3 expression.

    Science.gov (United States)

    Fiil, Berthe Katrine; Petersen, Morten

    2011-10-01

    Signal transduction through MAPK cascades is essential for eukaryotic cell response to various extracellular stimuli, such as the induction of innate immune responses. Arabidopsis thaliana relies in particular on three of its 20 MAPKs, MPK3,-4,-6, for a proper immune response. Recently we showed that one MPK4-substrate, MKS1, is required for basal resistance against the virulent Pseudomonas syringae and the oomycete Hyaloperonospora arabidopsidis. Overexpression of MKS1 (35S-MKS1) led to increased resistance to the same pathogens but also to an increased susceptibility towards the fungi Botrytis cinerea. MKS1 interacts with the transcription factor WRKY33, which in turn controls the regulation of PAD3 and CYP71A13, two genes, required for proper resistance to B. cinerea. Therefore, we tested if the increased susceptibility towards B. cinerea from 35S-MKS1 was due to deregulation of WRKY33 targets. PAD3 and CYP71A13 expression is similar in 35S-MKS1 and WT after B. cinerea treatment suggesting another mechanism controls 35S-MKS1 susceptibility.

  20. Variation among volatile profiles induced by Botrytis cinerea infection of tomato plants

    NARCIS (Netherlands)

    Jansen, R.M.C.

    2007-01-01

    Botrytis blight caused by the fungus Botrytis cinerea is probably the most common disease of greenhouse-grown crops like tomato. Botrytis blight in tomato plants is mainly detected by visual inspection or destructive biochemical and molecular determinations. These methods are time consuming and not

  1. Towards consumer-friendly cisgenic strawberries which are less susceptible to Botrytis cinerea

    NARCIS (Netherlands)

    Schaart, J.G.

    2004-01-01

    This thesis describes the development of genetically modified (GM) strawberries which are less susceptible to fruit rot caused by the fungus Botrytis cinerea. To achieve Botrytis resistance, a polygalacuronase inhibiting protein (PGIP) gene has been isolation from strawberry and was characterised. I

  2. Inhibitory effect of boron against Botrytis cinerea on table grapes and its possible mechanisms of action.

    Science.gov (United States)

    Qin, Guozheng; Zong, Yuanyuan; Chen, Qiling; Hua, Donglai; Tian, Shiping

    2010-03-31

    Boron, an essential plant micronutrient, was effective in the form of potassium tetraborate for control of postharvest gray mold caused by Botrytis cinerea on table grapes stored at room temperature or at 0 degrees C. The inhibition of fruit decay was closely correlated with boron concentrations and partially influenced by pH value of the solution. Boron strongly inhibited spore germination, germ tube elongation, and mycelial spread of B. cinerea in the culture medium. Application of boron at 1% caused the appearance of abnormal spores (disrupted) in some cases. By using propidium iodide fluorescent staining, loss of membrane integrity in B. cinerea was observed after boron treatment. Furthermore, boron led to the leakage of cellular constituents (soluble proteins and carbohydrates) from hyphae of B. cinerea. These data suggest that the mechanisms by which boron decreased gray mold decay of table grapes may be directly related to the disruption effect of boron on cell membrane of the fungal pathogen that resulted in the breakdown of the cell membrane and loss of cytoplasmic materials from the hyphae.

  3. BcSUN1, a B. cinerea SUN-Family Protein, Is Involved in Virulence

    Science.gov (United States)

    Pérez-Hernández, Alicia; González, Mario; González, Celedonio; van Kan, Jan A. L.; Brito, Nélida

    2017-01-01

    BcSUN1 is a glycoprotein secreted by Botrytis cinerea, an important plant pathogen that causes severe losses in agriculture worldwide. In this work, the role of BcSUN1 in different aspects of the B. cinerea biology was studied by phenotypic analysis of Bcsun1 knockout strains. We identified BcSUN1 as the only member of the Group-I SUN family of proteins encoded in the B. cinerea genome, which is expressed both in axenic culture and during infection. BcSUN1 is also weakly attached to the cellular surface and is involved in maintaining the structure of the cell wall and/or the extracellular matrix. Disruption of the Bcsun1 gene produces different cell surface alterations affecting the production of reproductive structures and adhesion to plant surface, therefore reducing B. cinerea virulence. BcSUN1 is the first member of the SUN family reported to be involved in the pathogenesis of a filamentous fungus. PMID:28163701

  4. Differential predation on tadpoles influences the potential effects of hybridization between Hyla cinerea and Hyla gratiosa

    Science.gov (United States)

    Gunzburger, M.S.

    2005-01-01

    Long-term effects of hybridization and introgression are influenced by performance of hybrids in habitats of parental species. The treefrogs Hyla cinerea and Hyla gratiosa, which typically breed in permanent and temporary habitats, respectively, have occasionally hybridized throughout the Southeastern United States. To predict in which of the parental habitats effects of hybridization might be strongest, I performed experiments to evaluate predation on tadpoles of H. cinerea, H. gratiosa, and F1 hybrids with predators typical of the breeding habitats of the parental species. Hybrid tadpoles had lower survival with sunfish than odonate naiad (dragonfly) predators and tended to increase hiding behavior in response to sunfish predation. Tadpoles of H. gratiosa also had higher survival with odonates than sunfish, but H. cinerea had similar survival with both predator types. These results suggest that hybrids are most likely to survive and return to breed in temporary habitats used by H. gratiosa. Thus, hybridization and introgression might be more likely to have adverse effects on populations of H. gratiosa than H. cinerea. Copyright 2005 Society for the Study of Amphibians and Reptiles.

  5. Determination of fungicide resistance in Botrytis cinerea from strawberry in the Central Coast Region of California

    Science.gov (United States)

    A study was conducted in 2013 to investigate the occurrence of fungicide resistance in Botrytis cinerea populations in California’s northern strawberry growing region; specifically in Watsonville and Salinas. In mid-May, 59 samples consisting of a single diseased fruit or plant part with gray mold s...

  6. Ulocladium atrum 385: Een veelbelovende kandidaat voor de biologische bestrijding van Botrytis cinerea

    NARCIS (Netherlands)

    Köhl, J.; Molhoek, W.M.L.

    2002-01-01

    De schimmel Ulocladium atrum is geselecteerd als een antagonist van Botrytis cinerea. De ecologische eigenschappen van deze antagonist en de toepassingen op bovengrondse plantendelen worden in dit artikel beschreven. Gegevens bij de bijgaande figuren: 1) Effect van Ucladium atrum op de grauwe schimm

  7. Characterization and inhibitory activity of chitosan on hyphae growth and morphology of Botrytis cinerea plant pathogen

    Directory of Open Access Journals (Sweden)

    Sebastião Silva Junior

    2014-07-01

    Full Text Available Summary. Low and high molecular weight chitosan were tested in different concentrations and growth times with the aim to evaluate the inhibitory activity against Botrytis cinerea, a very important plant pathogen. Tested chitosans were characterized by vibratory spectroscopy and elementary analyzes to determine the deacetylation degree. In addiction molar mass was estimated by viscosity measuring. Scanning electron microscopy was utilized for antimicrobial activity observation. Results showed that both chitosans markedly inhibited fungal growth, which was effected by incubation time and chitosan concentration. Scanning electron microscopy observations revealed that chitosan induced changes in surface morphology. The present study show that chitosan is capable of inhibit the growth and cause serious damage to the cell structure of the B. cinerea, as well as have the ability to form an impervious layer around the cell. Therefore, chitosan could be considered as a potential alternative for synthetic fungicides.Industrial relevance. Ultrastructural analysis showed that chitosan is capable of causing serious damage to the cell structure of the B. cinerea, as well as have the ability to form an impervious layer around the cell. Chitosan could inhibit the growth of B. cinerea in vitro and consequently may be considered as a potential alternative in replacement of synthetic fungicides.Keywords. biopolymer; chitosan; antifungal activity; fungal morphology; electron microscopy

  8. Postharvest Control of Botrytis cinerea and Monilinia fructigena in Apples by Gamma Irradiation Combined with Fumigation.

    Science.gov (United States)

    Cheon, Wonsu; Kim, Young Soo; Balaraju, Kotnala; Kim, Bong-Su; Lee, Byeong-Ho; Jeon, Yongho

    2016-08-01

    To extend the shelf life of apples in South Korea, we evaluated the effect of gamma irradiation alone or gamma irradiation combined with fumigation on the control of postharvest decay caused by Botrytis cinerea and Monilinia fructigena. An irradiation dose of 1.0 kGy caused the maximal inhibition of B. cinerea and M. fructigena spore germination. The gamma irradiation dose required to reduce the spore germination by 90% was 0.76 and 0.78 kGy for B. cinerea and M. fructigena, respectively. Inhibition of conidial germination of both fungal pathogens occurred at a greater level at the doses of 0.2 to 1.0 kGy compared with the nontreated control; 0.2 kGy caused 90.5 and 73.9% inhibition of B. cinerea and M. fructigena, respectively. Treatment in vitro with the ecofriendly fumigant ethanedinitrile had a greater effect compared with the nontreated control. The in vitro antifungal effects of the gamma irradiation and fumigation treatments allowed us to further study the effects of the combined treatments. Interestingly, when irradiation was combined with fumigation, the percentage of disease inhibition increased more at lower (<0.4 kGy) than at higher doses of irradiation, suggesting that the combined treatments reduced the necessary irradiation dose in phytosanitary irradiation processing under storage conditions.

  9. Ciprofloxacin treatment of bacterial peritonitis associated with chronic ambulatory peritoneal dialysis caused by Neisseria cinerea.

    Science.gov (United States)

    Taegtmeyer, M; Saxena, R; Corkill, J E; Anijeet, H; Parry, C M

    2006-08-01

    Bacterial peritonitis is a well-recognized complication of chronic ambulatory peritoneal dialysis (CAPD) in patients with end-stage renal failure. We present a case of peritonitis due to an unusual pathogen, Neisseria cinerea, unresponsive to the standard intraperitoneal (i.p.) vancomycin and gentamicin, which responded rapidly to oral ciprofloxacin.

  10. Neisseria cinerea isolates can adhere to human epithelial cells by type IV pilus-independent mechanisms.

    Science.gov (United States)

    Wörmann, Mirka E; Horien, Corey L; Johnson, Errin; Liu, Guangyu; Aho, Ellen; Tang, Christoph M; Exley, Rachel M

    2016-03-01

    In pathogenic Neisseria species the type IV pili (Tfp) are of primary importance in host-pathogen interactions. Tfp mediate initial bacterial attachment to cell surfaces and formation of microcolonies via pilus-pilus interactions. Based on genome analysis, many non-pathogenic Neisseria species are predicted to express Tfp, but aside from studies on Neisseria elongata, relatively little is known about the formation and function of pili in these organisms. Here, we have analysed pilin expression and the role of Tfp in Neisseria cinerea. This non-pathogenic species shares a close taxonomic relationship to the pathogen Neisseria meningitidis and also colonizes the human oropharyngeal cavity. Through analysis of non-pathogenic Neisseria genomes we identified two genes with homology to pilE, which encodes the major pilin of N. meningitidis. We show which of the two genes is required for Tfp expression in N. cinerea and that Tfp in this species are required for DNA competence, similar to other Neisseria. However, in contrast to the meningococcus, deletion of the pilin gene did not impact the association of N. cinerea to human epithelial cells, demonstrating that N. cinerea isolates can adhere to human epithelial cells by Tfp-independent mechanisms.

  11. Grape Berry Colonization and Biological Control of Botrytis cinerea by Indigenous Vineyard Yeasts

    Science.gov (United States)

    Botrytis bunch rot, caused by Botrytis cinerea, is the most important disease of grape berries, especially during transportation and storage. Biological control is a potential means of postharvest management of Botrytis bunch rot. The study was aimed at testing the hypothesis that antagonistic yeast...

  12. Botrytis cinerea endopolygalacturonase genes are differentially expressed in various plant tissues

    NARCIS (Netherlands)

    Have, ten A.; Oude Breuil, W.; Wubben, J.P.; Visser, J.; Kan, van J.A.L.

    2001-01-01

    Botrytis cinerea, the causal agent of blight, rot, and gray mold on many plant species, secretes various endopolygalacturonases during all stages of infection. The expression pattern of the encoding genes (Bcpg 1-6) was studied on four hosts: tomato, broad bean, apple, and courgette (also known as z

  13. AISLAMIENTO Y EVALUACIÓN IN VITRO DE ANTAGONISTAS DE Botrytis cinerea EN MORA

    Directory of Open Access Journals (Sweden)

    Jos\\u00E9 Alonso Calvo-Araya

    2012-01-01

    Full Text Available El objetivo de este estudio fue determinar la capacidad antagónica de hongos a Botrytis cinerea en el cultivo de la mora en Costa Rica. Durante el primer semestre del 2009 se aislaron 35 hongos filamentosos habitantes del carpoplano de frutos de mora, de los cuales seis cepas de Trichoderma fueron seleccionadas para su evaluación in vitro contra B. cinerea por medio de la técnica de cultivos duales. En la evaluación se determinó la competencia por sustrato y el efecto antibiótico. Para evaluar la competencia por sustrato se utilizó la escala de Bell y en el caso del efecto antibiótico se calculó el porcentaje de inhibición del crecimiento. Todas las cepas evaluadas compitieron eficientemente por sustrato contra B. cinerea, destacaron los aislamientos Lu13 y Lu15, que alcanzaron el grado I en la escala usada, donde el antagonista sobrepasó y creció sobre el patógeno cubriendo el 100% del medio de cultivo. Los restantes aislamientos de Trichoderma alcanzaron el grado II de antagonismo. En cuanto al efecto antibiótico, todos los aislamientos inhibieron el crecimiento micelial de B. cinerea, cuatro de ellos alcanzaron un valor mayor al 80% al ser evaluados en condiciones in vitro.

  14. Systematic notes on Asian birds. 39. The correct name for the Mangrove Whistler Pachycephala cinerea (Blyth)

    NARCIS (Netherlands)

    Walters, M.P.

    2003-01-01

    The correct scientific name for the Mangrove Whistler is Pachycephala cinerea (Blyth, 1847) and not Pachycephala grisola (Blyth, 1843). The proposal for change by Mukherjee (1970) was based on a purported type specimen of the latter, which cannot qualify as a type.

  15. The toolbox of Trichoderma spp. in the biocontrol of Botrytis cinerea disease.

    Science.gov (United States)

    Vos, Christine M F; De Cremer, Kaat; Cammue, Bruno P A; De Coninck, Barbara

    2015-05-01

    Botrytis cinerea is a necrotrophic fungal pathogen causing disease in many plant species, leading to economically important crop losses. So far, fungicides have been widely used to control this pathogen. However, in addition to their detrimental effects on the environment and potential risks for human health, increasing fungicide resistance has been observed in the B. cinerea population. Biological control, that is the application of microbial organisms to reduce disease, has gained importance as an alternative or complementary approach to fungicides. In this respect, the genus Trichoderma constitutes a promising pool of organisms with potential for B. cinerea control. In the first part of this article, we review the specific mechanisms involved in the direct interaction between the two fungi, including mycoparasitism, the production of antimicrobial compounds and enzymes (collectively called antagonism), and competition for nutrients and space. In addition, biocontrol has also been observed when Trichoderma is physically separated from the pathogen, thus implying an indirect systemic plant defence response. Therefore, in the second part, we describe the consecutive steps leading to induced systemic resistance (ISR), starting with the initial Trichoderma-plant interaction and followed by the activation of downstream signal transduction pathways and, ultimately, the defence response resulting in ISR (ISR-prime phase). Finally, we discuss the ISR-boost phase, representing the effect of ISR priming by Trichoderma spp. on plant responses after additional challenge with B. cinerea.

  16. Deficiencies in jasmonate-mediated plant defense reveal quantitative variation in Botrytis cinerea pathogenesis.

    Directory of Open Access Journals (Sweden)

    Heather C Rowe

    2010-04-01

    Full Text Available Despite the described central role of jasmonate signaling in plant defense against necrotrophic pathogens, the existence of intraspecific variation in pathogen capacity to activate or evade plant jasmonate-mediated defenses is rarely considered. Experimental infection of jasmonate-deficient and jasmonate-insensitive Arabidopsis thaliana with diverse isolates of the necrotrophic fungal pathogen Botrytis cinerea revealed pathogen variation for virulence inhibition by jasmonate-mediated plant defenses and induction of plant defense metabolites. Comparison of the transcriptional effects of infection by two distinct B. cinerea isolates showed only minor differences in transcriptional responses of wild-type plants, but notable isolate-specific transcript differences in jasmonate-insensitive plants. These transcriptional differences suggest B. cinerea activation of plant defenses that require plant jasmonate signaling for activity in response to only one of the two B. cinerea isolates tested. Thus, similar infection phenotypes observed in wild-type plants result from different signaling interactions with the plant that are likely integrated by jasmonate signaling.

  17. The homeobox BcHOX8 gene in Botrytis cinerea regulates vegetative growth and morphology.

    Directory of Open Access Journals (Sweden)

    Zsuzsanna Antal

    Full Text Available Filamentous growth and the capacity at producing conidia are two critical aspects of most fungal life cycles, including that of many plant or animal pathogens. Here, we report on the identification of a homeobox transcription factor encoding gene that plays a role in these two particular aspects of the development of the phytopathogenic fungus Botrytis cinerea. Deletion of the BcHOX8 gene in both the B. cinerea B05-10 and T4 strains causes similar phenotypes, among which a curved, arabesque-like, hyphal growth on hydrophobic surfaces; the mutants were hence named Arabesque. Expression of the BcHOX8 gene is higher in conidia and infection cushions than in developing appressorium or mycelium. In the Arabesque mutants, colony growth rate is reduced and abnormal infection cushions are produced. Asexual reproduction is also affected with abnormal conidiophore being formed, strongly reduced conidia production and dramatic changes in conidial morphology. Finally, the mutation affects the fungus ability to efficiently colonize different host plants. Analysis of the B. cinerea genome shows that BcHOX8 is one member of a nine putative homeobox genes family. Available gene expression data suggest that these genes are functional and sequence comparisons indicate that two of them would be specific to B. cinerea and its close relative Sclerotinia sclerotiorum.

  18. Partial stem and leaf resistance against the fungal pathogen Botrytis cinerea in wild relatives of tomato

    NARCIS (Netherlands)

    Have, ten A.; Berloo, van R.; Lindhout, P.; Kan, van J.A.L.

    2007-01-01

    Tomato (Solanum lycopersicum) is one of many greenhouse crops that can be infected by the necrotrophic ascomycete Botrytis cinerea. Commercial cultivation of tomato is hampered by the lack of resistance. Quantitative resistance has been reported in wild tomato relatives, mostly based on leaf assays.

  19. Cloning, expression and characteristics of a novel alkalistable and thermostable xylanase encoding gene (Mxyl retrieved from compost-soil metagenome.

    Directory of Open Access Journals (Sweden)

    Digvijay Verma

    Full Text Available BACKGROUND: The alkalistable and thermostable xylanases are in high demand for pulp bleaching in paper industry and generating xylooligosaccharides by hydrolyzing xylan component of agro-residues. The compost-soil samples, one of the hot environments, are expected to be a rich source of microbes with thermostable enzymes. METHODOLOGY/PRINCIPAL FINDINGS: Metagenomic DNA from hot environmental samples could be a rich source of novel biocatalysts. While screening metagenomic library constructed from DNA extracted from the compost-soil in the p18GFP vector, a clone (TSDV-MX1 was detected that exhibited clear zone of xylan hydrolysis on RBB xylan plate. The sequencing of 6.321 kb DNA insert and its BLAST analysis detected the presence of xylanase gene that comprised 1077 bp. The deduced protein sequence (358 amino acids displayed homology with glycosyl hydrolase (GH family 11 xylanases. The gene was subcloned into pET28a vector and expressed in E. coli BL21 (DE3. The recombinant xylanase (rMxyl exhibited activity over a broad range of pH and temperature with optima at pH 9.0 and 80°C. The recombinant xylanase is highly thermostable having T1/2 of 2 h at 80°C and 15 min at 90°C. CONCLUSION/SIGNIFICANCE: This is the first report on the retrieval of xylanase gene through metagenomic approach that encodes an enzyme with alkalistability and thermostability. The recombinant xylanase has a potential application in paper and pulp industry in pulp bleaching and generating xylooligosaccharides from the abundantly available agro-residues.

  20. Effects of indole-3-acetic acid on Botrytis cinerea isolates obtained from potted plants.

    Science.gov (United States)

    Martínez, J A; Valdés, R; Gómez-Bellot, M J; Bañón, S

    2011-01-01

    We study the growth of different isolates of Botrytis cinerea collected from potted plants which were affected by Botrytis blight in southern Spain during recent years. These isolates, which show widely phenotypic differences when grown in vitro, are differentially affected by growth temperature, gibberellic acid applications and paclobutrazol, an efficient plant growth retardant and fungicide at the same time. In this work, we have evaluated the effect of the auxin indole-3-acetic acid (IAA) dose (0, 1, 10, and 100 mg/plate) on the growth of the collection of B. cinerea isolates obtained from the following potted plants: Cyclamen persicum, Hydrangea macrophylla, Lantona camara, and Lonicera japonica. B. cinerea produces indolacetic acid, but so far the precise biosynthetic pathway and some effects on this fungal species are still unclear, although recent studies have revealed an antifungal activity of IAA on several fungi, including B. cinerea isolated from harvested fruits. Mycelial growth curves and growth rates assessed from difference in colony areas during the both linear and deceleration phase, conidiation (measured as time of appearance), conidia length (microm), and sclerotia production (number/plate) were evaluated in the isolates, which were grown at 26 degrees C on Petri dishes containing potato dextrose agar for up to 35 days. Mycelial growth curves fitted a typical kinetic equation of fungi grown on solid media. B. cinerea isolates showed a high degree of variability in their growth kinetics, depending on the isolate and auxin dose. This plant growth substance delayed mycelial growth during the linear phase in an isolate-dependent manner, thus isolates from C. persicum, H. macrophylla and L. camara were more affected by IAA than L. japonica. On the other hand, 100 mg of IAA was the critical dose to significantly reduce the growth rate in all isolates and to promote brown-striped hyphae development, especially in isolate from C. persicum. 10 and 100 mg

  1. Does release of encapsulated nutrients have an important role in the efficacy of xylanase in broilers?

    Science.gov (United States)

    Khadem, A; Lourenço, M; Delezie, E; Maertens, L; Goderis, A; Mombaerts, R; Höfte, M; Eeckhaut, V; Van Immerseel, F; Janssens, G P J

    2016-05-01

    The non-starch polysaccharides (NSPs) in cell walls can act as a barrier for digestion of intracellular nutrients. This effect is called "cage effect." Part of the success of fibrolytic enzymes in broiler feed is assumed to be attributed to cage effect reduction. Further, changes in viscosity and potential prebiotic action should also be considered. The aim of this study was to gain insight into the relative importance of the cage effect in xylanase efficacy in broilers. Using a 2×2 factorial design, 24 pens with 30 Ross 308 male chicks were fed corn-soy based diets consisting of normal and freeze-thawed (5 d at -18°C) corn, both with and without xylanase. The freeze-thaw method was used to eliminate the cage effect, whereas a corn-based diet was used to exclude viscosity effects. Body weights (BW), feed intake (FI), and feed conversion ratio (FCR) were determined at d 13, 26, and 39. A balance study was executed at the end of the growing phase. These birds were euthanized at d 34 (non-fasted) to determine the viscosity of digesta, blood metabolites, intestinal morphology, and microbiota composition. During the finisher period, there was a significant interaction between enzyme supplementation and freeze-thawing for FCR, in which FCR was improved by freeze-thawed corn and tended to be improved by normal corn+enzyme compared with the control group. The improvement in performance (finisher period) of freeze-thawed corn and xylanase coincided with increased gut absorption of glucose (based on postprandial plasma concentrations) and increased number of Clostridiumcluster IV in the caecum, and agreed with the higher gut villus height. In addition, xylanase inclusion significantly increased the postprandial plasma glycine and triglycerides concentration, and led to elevated bacterial gene copies of butyryl CoA:acetate CoA-transferase, suggesting a prebiotic effect of xylanase addition through more than just the cage effect reduction. The applied model managed to rule

  2. Biocontrol of Botrytis cinerea by successful introduction of Pantoea ananatis in the grapevine phyllosphere

    Directory of Open Access Journals (Sweden)

    Gasser F

    2012-12-01

    Full Text Available Florian Gasser,1 Massimiliano Cardinale,1 Barbara Schildberger,2 Gabriele Berg11Institute of Environmental Biotechnology, Graz University of Technology, Graz, Austria; 2Höhere Bundesanstalt und Bundesamt für Wein-und Obstbau, Klosterneuburg, AustriaBackground and aims: The fungus Botrytis cinerea is a common problem in viticulture and leads to serious losses in both yield and quality. The objective was to study the potential of the antagonist Pantoea ananatis BLBT1-08 for controlling this disease.Methods: Pathogen suppression by Pantoea treatments was investigated in different field trials and in detached leaf assays. The mode of action was studied by confocal laser scanning microscopy of treated grape leaves and by in vitro assays.Results: The introduction of P. ananatis BLBT1-08 in a 3-year field trial resulted in statistically significant reduction of disease symptoms. However, B. cinerea abundance, measured by quantitative real-time polymerase chain reaction of a B. cinerea specific gene, was not reduced when compared to non-treated, symptom-free leaves. A DsRed fluorescent protein labeled BLBT1-08 strain showed a high phyllosphere competence and competition on the leaf surface, but did not colonize the inner parts of plant tissue. Germination of B. cinerea was not inhibited by BLBT1-08 on the leaf, but mycelial growth and symptoms were suppressed without direct pathogen-antagonist contact. The antimicrobial activity was amino acid and temperature dependent.Conclusion: P. ananatis BLBT1-08 is a competitive and promising biocontrol agent for the control of B. cinerea and is highly effective at reducing disease incidence.Keywords: biological control, sustainable viticulture, antagonism

  3. Global and grain-specific accumulation of glycoside hydrolase family 10 xylanases in transgenic maize (Zea mays).

    Science.gov (United States)

    Gray, Benjamin N; Bougri, Oleg; Carlson, Alvar R; Meissner, Judy; Pan, Shihao; Parker, Matthew H; Zhang, Dongcheng; Samoylov, Vladimir; Ekborg, Nathan A; Michael Raab, R

    2011-12-01

    In planta expression of cell wall degrading enzymes is a promising approach for developing optimized biomass feedstocks that enable low-cost cellulosic biofuels production. Transgenic plants could serve as either an enzyme source for the hydrolysis of pretreated biomass or as the primary biomass feedstock in an autohydrolysis process. In this study, two xylanase genes, Bacillus sp. NG-27 bsx and Clostridium stercorarium xynB, were expressed in maize (Zea mays) under the control of two different promoters. Severe phenotypic effects were associated with xylanase accumulation in maize, including stunted plants and sterile grains. Global expression of these xylanases from the rice ubiquitin 3 promoter (rubi3) resulted in enzyme accumulation of approximately 0.01 mg enzyme per gram dry weight, or approximately 0.1% of total soluble protein (TSP). Grain-specific expression of these enzymes from the rice glutelin 4 promoter (GluB-4) resulted in higher-level accumulation of active enzyme, with BSX and XynB accumulating up to 4.0% TSP and 16.4% TSP, respectively, in shriveled grains from selected T0 plants. These results demonstrate the potential utility of the GluB-4 promoter for biotechnological applications. The phenotypic effects of xylanase expression in maize presented here demonstrate the difficulties of hemicellulase expression in an important crop for cellulosic biofuels production. Potential alternate approaches to achieve xylanase accumulation in planta without the accompanying negative phenotypes are discussed.

  4. The influence of some factors on β-1,4-xylanase activity of the filamentous fungus Trichoderma reesei QM9414

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    Alexandru Manoliu

    2012-03-01

    Full Text Available The mesophyllic fungus Trichoderma reesei (anamorph to Hypocrea jecorina is an important biotechnological tool, known for its ability to secrete large quantities of hydrolytic enzymes. Renewable biomass, such as agricultural and forest wastes are used to produce microbial enzymes in various industrial processes such as food, feed and bioethanol industries. In raw biomass materials, such as wheat straws, barley straws and maize stalks, the main polysaccharide is cellulose which is closely associated with hemicelluloses like xylan, manan and xyloguclan. In consequence, the hydrolysis of these materials requires the concerted action of several enzymes, namely cellulases and xylanases. Endo-xylanase (endo-1,4--xylanase, EC 3.2.1.8 is the key enzyme involved in xylan hydrolysis, the mainhemicellulosic component of plant cell walls. The metabolic activity and enzyme productivity of Trichoderma reesei isinfluenced by various environmental conditions. In this context, we analysed the effect of pH, cultivation period, thenature of the substrate used and the nitrogen source on enzymatic activity. The maximum xylanase yield was recorded at a initial pH of 4 (116.189 IU/ml for barley and 5 for wheat (88.578 IU/ml, respectively maize (116.583 IU/ml. The bestsubstrate for endo-xylanase activity was maize stalks (90.446 IU/ml at a a concentration of 30g/L.

  5. Combination of Xylanase and Debranching Enzymes Specific to Wheat Arabinoxylan Improve the Growth Performance and Gut Health of Broilers.

    Science.gov (United States)

    Lei, Zhao; Shao, Yuxin; Yin, Xiaonan; Yin, Dafei; Guo, Yuming; Yuan, Jianmin

    2016-06-22

    Arabinoxylan (AX) is the major antinutritional factor of wheat. This study evaluated the synergistic effects of xylanase and debranching enzymes (arabinofuranosidase [ABF] and feruloyl esterase [FAE]) on AX. During in vitro tests, the addition of ABF or FAE accelerated the hydrolysis of water-soluble AX (WE-AX) and water-insoluble AX (WU-AX) and produced more xylan oligosaccharides (XOS) than xylanase alone. XOS obtained from WE-AX stimulated greater proliferation of Lactobacillus brevis and Bacillus subtilis than did fructo-oligosaccharides (FOS) and glucose. During in vivo trials, xylanase increased the average daily growth (ADG), decreased the feed-conversion ratio (FCR), and reduced the digesta viscosity of jejunum and intestinal lesions of broilers fed a wheat-based diet on day 36. ABF or FAE additions further improved these effects. Broilers fed a combination of xylanase, ABF, and FAE exhibited the best growth. In conclusion, the synergistic effects among xylanase, ABF, and FAE increased AX degradation, which improve the growth performance and gut health of broilers.

  6. Xylanases, Cellulases, and Acid Protease Produced by Stenocarpella maydis Grown in Solid-state and Submerged Fermentation

    Directory of Open Access Journals (Sweden)

    Edna María Hernández-Domínguez

    2014-03-01

    Full Text Available Activity levels of extracellular hydrolytic enzymes produced by Stenocarpella maydis, a fungal pathogen of maize, have so far not been reported. Production of xylanase, cellulase, and acid protease by this ascomycete using different culture media in solid-state and submerged fermentation was studied. In solid-state fermentation, polyurethane foam was used as an inert support, and corncob, corn leaves, and broken corn were used as biodegradable supports. The highest xylanase activity was produced in the medium with xylan in both fermentation systems, reaching 18,020 U/L and 19,266 U/L for submerged and solid-state fermentation, respectively. Cellulase production was observed only in the culture medium with carboxymethylcellulose, obtaining values of 7,872 U/L in submerged fermentation and 9,439 U/L in solid-state fermentation. The acid protease was produced only in minimal medium with glucose in acidic pH, reaching the highest levels of activity in SSF (806 U/L. The corncob was the best biodegradable support for the production of xylanases and acid protease. Two isoenzymes of xylanase and cellulase were observed in both fermentation systems, and three isoenzymes of xylanase were produced on the biodegradable supports.

  7. Production of Xylanase from Arthrobacter sp. MTCC 6915 Using Saw Dust As Substrate under Solid State Fermentation

    Directory of Open Access Journals (Sweden)

    Sevanan Murugan

    2011-01-01

    Full Text Available Saw dust was used as substrate for xylanase production from Arthrobacter sp. MTCC 6915. The study of period of incubation, temperature, pH, carbon, and nitrogen sources for xylanase production was optimized. Xylanase production was found to be optimum at an incubation period of 96 hrs (117.0 U/mL, temperature 30°C (105.0 U/mL, and pH 9.0 (102.9 U/mL. The results showed that the xylanase production was found to be higher in the presence of carboxymethylcellulose (176.4 U/mL and dextrose (126.0 U/mL. It was also observed that peptone (170.1 U/mL and beef extract (161.7 U/mL supported maximum xylanase production.The enzyme was characterized and found to be fairly active at pH 9 (764.4 U/mL and temperature 60°C (819 U/mL. Even in the present study, a major difference in the production temperature (30°C and optimal temperature (60°C of the enzyme activity was observed. However, the pH of the production media and the enzyme activity were found to be the same (pH 9.

  8. Bioprocess and biotecnology: effect of xylanase from Aspergillus niger and Aspergillus flavus on pulp biobleaching and enzyme production using agroindustrial residues as substract.

    Science.gov (United States)

    de Alencar Guimaraes, Nelciele Cavalieri; Sorgatto, Michele; Peixoto-Nogueira, Simone de Carvalho; Betini, Jorge Henrique Almeida; Zanoelo, Fabiana Fonseca; Marques, Maria Rita; de Moraes Polizeli, Maria de Lourdes Teixeira; Giannesi, Giovana C

    2013-01-01

    This study compares two xylanases produced by filamentous fungi such as A. niger and A. flavus using agroindustrial residues as substract and evaluated the effect of these enzymes on cellulose pulp biobleaching process. Wheat bran was the best carbon source for xylanase production by A. niger and A. flavus. The production of xylanase was 18 and 21% higher on wheat bran when we compare the xylanase production with xylan. At 50°C, the xylanase of A. niger retained over 85% activity with 2 h of incubation, and A. flavus had a half-life of more than 75 minutes. At 55°C, the xylanase produced by A. niger showed more stable than from A. flavus showing a half-life of more than 45 minutes. The xylanase activity of A. niger and A. flavus were somehow protected in the presence of glycerol 5% when compared to the control (without additives). On the biobleaching assay it was observed that the xylanase from A. flavus was more effective in comparison to A. niger. The kappa efficiency corresponded to 36.32 and 25.93, respectively. That is important to emphasize that the cellulase activity was either analyzed and significant levels were not detected, which explain why the viscosity was not significantly modified.

  9. Development and evaluation of a novel and rapid detection assay for Botrytis cinerea based on loop-mediated isothermal amplification.

    Science.gov (United States)

    Duan, Ya-Bing; Ge, Chang-Yan; Zhang, Xiao-Ke; Wang, Jian-Xin; Zhou, Ming-Guo

    2014-01-01

    Botrytis cinerea is a devastating plant pathogen that causes grey mould disease. In this study, we developed a visual detection method of B. cinerea based on the Bcos5 sequence using loop-mediated isothermal amplification (LAMP) with hydroxynaphthol blue dye (HNB). The LAMP reaction was optimal at 63 °C for 45 min. When HNB was added prior to amplification, samples with B. cinerea DNA developed a characteristic sky blue color after the reaction but those without DNA or with DNA of other plant pathogenic fungi did not. Results of HNB staining method were reconfirmed when LAMP products were subjected to gel electrophoresis. The detection limit of this LAMP assay for B. cinerea was 10(-3) ng µL(-1) of genomic DNA per reaction, which was 10-fold more sensitive than conventional PCR (10(-2) ng µL(-1)). Detection of the LAMP assay for inoculum of B. cinerea was possible in the inoculated tomato and strawberry petals. In the 191 diseased samples, 180 (94.2%) were confirmed as positive by LAMP, 172 (90.1%) positive by the tissue separation, while 147 (77.0%) positive by PCR. Because the LAMP assay performed well in aspects of sensitivity, specificity, repeatability, reliability, and visibility, it is suitable for rapid detection of B. cinerea in infected plant materials prior to storage and during transportation, such as cut flowers, fruits and vegetables.

  10. Isolation and characteristics of protocatechuic acid from Paenibacillus elgii HOA73 against Botrytis cinerea on strawberry fruits.

    Science.gov (United States)

    Nguyen, Xuan Hoa; Naing, Kyaw Wai; Lee, Young Seong; Moon, Jae Hak; Lee, Jeong Hyun; Kim, Kil Yong

    2015-05-01

    This study was undertaken to describe purification, identification, and characteristics of protocatechuic acid (PCA) isolated for the first time from Paenibacillus elgii HOA73 against Botrytis cinerea (the cause of gray mold disease on strawberry fruit). PCA was purified by different chromatographic techniques and identified as PCA (3,4-dihydroxybenzoic acid) by nuclear magnetic resonance and liquid chromatography-mass spectrometry analyses. PCA displayed potent antifungal activity against B. cinerea and Rhizoctonia solani. However, the antifungal activities were not sufficient to inhibit mycelial growth of Phytophthora capsici and Fusarium oxysporum. The minimum inhibitory concentration of PCA to inhibit any visible mycelial growth of both B. cinerea and R. solani was 64 µg ml(-1) . Most B. cinerea conidia displayed altered shape and absence of germination, or were degraded after treatment with 50 and 100 µg ml(-1) PCA, respectively. Moreover, gray mold formation on strawberry fruit was almost or completely inhibited by these PCA concentrations 7 days following infection with B. cinerea conidia, respectively. PCA may be a promising alternative to chemical fungicides as a potential biofungicide to prevent growth of B. cinerea in strawberry fruit disease management.

  11. Impact of the Botrytis cinerea strain and metabolism on (-)-geosmin production by Penicillium expansum in grape juice.

    Science.gov (United States)

    La Guerche, Stéphane; De Senneville, Laure; Blancard, Dominique; Darriet, Philippe

    2007-10-01

    Geosmin, an off-flavour of some rotten grapes, has been implicated in wine defects. Botrytis cinerea and Penicillium expansum were the most common among the numerous microorganisms isolated from rotten grapes. P. expansum produces geosmin on model media but not healthy grape juice. However, geosmin synthesis by P. expansum was demonstrated in grape juice and on crushed grapes that had been pre-cultured with certain B. cinerea strains. 34 out of 156 B. cinerea strains ([bot +] phenotype) isolated from the centre of grape bunches were able to induce high geosmin production, up to 494 ng/l, by P. expansum in grape juice. A study of the impact of grape juice composition on geosmin synthesis by P. expansum revealed the importance of nitrogen composition, particularly amino-acid deficiency. Metabolism of amino acids by B. cinerea was shown to be favourable to geosmin synthesis by P. expansum. However, the amino-acid and ammonium concentrations in grape juices pre-cultured with B. cinerea [bot -] and [bot +] strains were very similar implying that other factors are involved as well. Indeed, an ethanol-precipitable fraction, probably a polysaccharide, synthesized by B. cinerea [bot -], but not [bot +] strains, inhibited geosmin production by P. expansum.

  12. Development and evaluation of a novel and rapid detection assay for Botrytis cinerea based on loop-mediated isothermal amplification.

    Directory of Open Access Journals (Sweden)

    Ya-Bing Duan

    Full Text Available Botrytis cinerea is a devastating plant pathogen that causes grey mould disease. In this study, we developed a visual detection method of B. cinerea based on the Bcos5 sequence using loop-mediated isothermal amplification (LAMP with hydroxynaphthol blue dye (HNB. The LAMP reaction was optimal at 63 °C for 45 min. When HNB was added prior to amplification, samples with B. cinerea DNA developed a characteristic sky blue color after the reaction but those without DNA or with DNA of other plant pathogenic fungi did not. Results of HNB staining method were reconfirmed when LAMP products were subjected to gel electrophoresis. The detection limit of this LAMP assay for B. cinerea was 10(-3 ng µL(-1 of genomic DNA per reaction, which was 10-fold more sensitive than conventional PCR (10(-2 ng µL(-1. Detection of the LAMP assay for inoculum of B. cinerea was possible in the inoculated tomato and strawberry petals. In the 191 diseased samples, 180 (94.2% were confirmed as positive by LAMP, 172 (90.1% positive by the tissue separation, while 147 (77.0% positive by PCR. Because the LAMP assay performed well in aspects of sensitivity, specificity, repeatability, reliability, and visibility, it is suitable for rapid detection of B. cinerea in infected plant materials prior to storage and during transportation, such as cut flowers, fruits and vegetables.

  13. Gene Overexpression and RNA Silencing Tools for the Genetic Manipulation of the S-(+-Abscisic Acid Producing Ascomycete Botrytis cinerea

    Directory of Open Access Journals (Sweden)

    Zhong-Tao Ding

    2015-05-01

    Full Text Available The phytopathogenic ascomycete Botrytis cinerea produces several secondary metabolites that have biotechnical significance and has been particularly used for S-(+-abscisic acid production at the industrial scale. To manipulate the expression levels of specific secondary metabolite biosynthetic genes of B. cinerea with Agrobacterium tumefaciens-mediated transformation system, two expression vectors (pCBh1 and pCBg1 with different selection markers and one RNA silencing vector, pCBSilent1, were developed with the In-Fusion assembly method. Both expression vectors were highly effective in constitutively expressing eGFP, and pCBSilent1 effectively silenced the eGFP gene in B. cinerea. Bcaba4, a gene suggested to participate in ABA biosynthesis in B. cinerea, was then targeted for gene overexpression and RNA silencing with these reverse genetic tools. The overexpression of bcaba4 dramatically induced ABA formation in the B. cinerea wild type strain Bc-6, and the gene silencing of bcaba4 significantly reduced ABA-production in an ABA-producing B. cinerea strain.

  14. Studies on Mutation Breeding of High-Yielding Xylanase Strains by Low-Energy Ion Beam Implantation

    Institute of Scientific and Technical Information of China (English)

    LI Shichang; YAO Jianming; YU Zhengling

    2007-01-01

    As a new mutagenetic method,low-energy ion implantation has been used widely in many research areas in recent years.In order to obtain some industrial strains with high xylanase yield,the wild type strain Aspergillus niger A3 was mutated by means of nitrogen ions implantation (10 keV,2.6 × 1014~1.56 × 1015 ions/cm2) and a mutant N212 was isolated subsequently.However,it was found that the initial screening means of the high-yielding xylanase strains such as transparent halos was unfit for first screening.Compared with that of the wild type strain,xylanase production of the mutant N212 was increased from 320 IU/ml to 610 IU/ml,and the optimum fermentation temperature was increased from 28 ℃ to 30 ℃.

  15. Kinetics and substrate selectivity of a Triticum aestivum xylanase inhibitor (TAXI) resistant D11F/R122D variant of Bacillus subtilis XynA xylanase

    DEFF Research Database (Denmark)

    Rasmussen, Louise Enggaard; Sørensen, Jens F.; Meyer, Anne S.

    2010-01-01

    ), water-unextractable arabinoxylan (WUAX), birchwood xylan, and wheat bran. Both the BsX and the BsX(mut) catalyzed the release of xylo-oligosaccharides with higher degree of polymerization from WUAX than from WEAX. At equimolar addition levels the activity of the BsX(mut) was lower than that of the Bs......X with respect to both the initial rate and the product yields obtained after prolonged reaction on the xylan substrates. The calculated substrate selectivity factors indicated that the BsX and the BsX(mut) both had higher catalytic rate on WUAX than on WEAX. Addition of a 100:1 (TAXI:xylanase) molar ratio...

  16. Xylanase production by Penicillium canescens on soya oil cake in solid-state fermentation.

    Science.gov (United States)

    Antoine, Assamoi Allah; Jacqueline, Destain; Thonart, Philippe

    2010-01-01

    There is an increasing interest for the organic residues from various sectors of agriculture and industries over the past few decades. Their application in the field of fermentation technology has resulted in the production of bulk chemicals and value-added products such as amino acid, enzymes, mushroom, organic acids, single-cell protein, biologically active secondary metabolites, etc. (Ramachandran et al., Bioresource Technology 98:2000-2009, 2007). In this work, the production of extracellular xylanase by the fungus Penicillium canescens was investigated in solid-state fermentation using five agro-industrial substrates (soya oil cake, soya meal, wheat bran, whole wheat bran, and pulp beet). The best substrate was the soya oil cake. In order to optimize the production, the most effective cultivation conditions were investigated in Erlenmeyer flasks and in plastic bags with 5 and 100 g of soya oil cake, respectively. The initial moisture content, initial pH, and temperature of the culture affected the xylanase synthesis. The optimal fermentation medium was composed by soya oil cake crushed to 5 mm supplemented with 3% and 4% (w/w) of casein peptone and Na(2)HPO(4) x 2H(2)O. After 7 days of incubation at 30 degrees C and under 80% of initial moisture, a xylanase production level of 18,895 +/- 778 U/g (Erlenmeyer flasks) and 9,300 +/- 589 U/g (plastic bags) was reached. The partially purified enzyme recovered by ammonium sulfate fractionation was completely stable at freezing and refrigeration temperatures up to 6 months and reasonably stable at room temperature for more than 3 months.

  17. Biorefining of wood: combined production of ethanol and xylanase from waste fiber sludge.

    Science.gov (United States)

    Cavka, Adnan; Alriksson, Björn; Rose, Shaunita H; van Zyl, Willem H; Jönsson, Leif J

    2011-08-01

    The possibility to utilize fiber sludge, waste fibers from pulp mills and lignocellulose-based biorefineries, for combined production of liquid biofuel and biocatalysts was investigated. Without pretreatment, fiber sludge was hydrolyzed enzymatically to monosaccharides, mainly glucose and xylose. In the first of two sequential fermentation steps, the fiber sludge hydrolysate was fermented to cellulosic ethanol with the yeast Saccharomyces cerevisiae. Although the final ethanol yields were similar, the ethanol productivity after 9.5 h was 3.3 g/l/h for the fiber sludge hydrolysate compared with only 2.2 g/l/h for a reference fermentation with similar sugar content. In the second fermentation step, the spent fiber sludge hydrolysate (the stillage obtained after distillation) was used as growth medium for recombinant Aspergillus niger expressing the xylanase-encoding Trichoderma reesei (Hypocrea jecorina) xyn2 gene. The xylanase activity obtained with the spent fiber sludge hydrolysate (8,500 nkat/ml) was higher than that obtained in a standard medium with similar monosaccharide content (1,400 nkat/ml). Analyses based on deglycosylation with N-glycosidase F suggest that the main part of the recombinant xylanase was unglycosylated and had molecular mass of 20.7 kDa, while a minor part had N-linked glycosylation and molecular mass of 23.6 kDa. Chemical analyses of the growth medium showed that important carbon sources in the spent fiber sludge hydrolysate included xylose, small aliphatic acids, and oligosaccharides. The results show the potential of converting waste fiber sludge to liquid biofuel and enzymes as coproducts in lignocellulose-based biorefineries.

  18. CLONING, PURIFICATION AND CHARACTERIZATION OF HALOTOLERANT XYLANASE FROM Geobacillus Thermodenitrificans C5

    Directory of Open Access Journals (Sweden)

    Muhammad Irfan

    2016-06-01

    Full Text Available High levels of extracellular xylanase activity (994.50 IU/ml produced by Geobacillus thermodenitrificans C5 originated gene was detected when it was expressed in E. coli BL21 host. Thermostable xylanase (GthC5Xyl was purified to homogeneity and showed a molecular mass of approximately 44 kDa according to SDS-PAGE. The specific activity of the purified GthC5Xyl was up to 1243.125IU/mg with a 9.89-fold purification. The activity of GthC5Xyl was stimulated by CoCl2, MnSO4, CuSO4, MnCl2 but was inhibited by FeSO4, Hg, CaSO4. GthC5Xyl showed resistant to SDS, Tween 20, Triton X-100, β- Mercaptoethanol, PMSF, DTT, NEM and DEPC, SDS, and EDTA. A greater affinity for oat spelt xylan was exhibited by GthC5Xyl with maximum enzymatic activity at 60°C and 6.0 pH. The activity portrayed by GthC5Xyl was found to be acellulytic with stability at high temperature (70°C-80°C and low pH (4.0 to 8.0. Xylobiose and xylopentose were the end products of proficient oat spelt xylanase hydrolysis by GthC5Xyl. High SDS resistance and broader stability of GthC5Xyl proves it to be worthy of impending application in numerous industrial processes especially textile, detergents and animal feed industry.

  19. Cloning and characterization of the first GH10 and GH11 xylanases from Rhizopus oryzae.

    Science.gov (United States)

    Xiao, Zhizhuang; Grosse, Stephan; Bergeron, Hélène; Lau, Peter C K

    2014-10-01

    The only available genome sequence for Rhizopus oryzae strain 99-880 was annotated to not encode any β-1,4-endoxylanase encoding genes of the glycoside hydrolase (GH) family 10 or 11. Here, we report the identification and cloning of two such members in R. oryzae strain NRRL 29086. Strain 29086 was one of several selected fungi grown on wheat or triticale bran and screened for xylanase activity among other hydrolytic actions. Its high activity (138 U/ml) in the culture supernatant led to the identification of two activity-stained proteins, designated Xyn-1 and Xyn-2 of respective molecular masses 32,000 and 22,000. These proteins were purified to electrophoretic homogeneity and characterized. The specific activities of Xyn-1 and Xyn-2 towards birchwood xylan were 605 and 7,710 U/mg, respectively. Kinetic data showed that the lower molecular weight Xyn-2 had a higher affinity (K m=3.2 ± 0.2 g/l) towards birchwood xylan than Xyn-1 by about 4-fold. The melting temperature (T m) of the two proteins, estimated to be in the range of 49.5-53.7 °C indicated that they are rather thermostable proteins. N-terminal and internal peptide sequences were obtained by chemical digestion of the purified xylanases to facilitate cloning, expression in Escherichia coli, and sequencing of the respective gene. The cloned Rhizopus xylanases were used to demonstrate release of xylose from flax shives-derived hemicellulose as model feedstock. Overall, this study expands the catalytic toolbox of GH10 and 11 family proteins that have applications in various industrial and bioproducts settings.

  20. Isothermal titration calorimetry and surface plasmon resonance allow quantifying substrate binding to different binding sites of Bacillus subtilis xylanase

    DEFF Research Database (Denmark)

    Cuyvers, Sven; Dornez, Emmie; Abou Hachem, Maher;

    2012-01-01

    Isothermal titration calorimetry and surface plasmon resonance were tested for their ability to study substrate binding to the active site (AS) and to the secondary binding site (SBS) of Bacillus subtilis xylanase A separately. To this end, three enzyme variants were compared. The first was a cat......Isothermal titration calorimetry and surface plasmon resonance were tested for their ability to study substrate binding to the active site (AS) and to the secondary binding site (SBS) of Bacillus subtilis xylanase A separately. To this end, three enzyme variants were compared. The first...

  1. Cellulose and hemicellulose-degrading enzymes in Fusarium commune transcriptome and functional characterization of three identified xylanases

    DEFF Research Database (Denmark)

    Yuhong, Huang; Busk, Peter Kamp; Lange, Lene

    2015-01-01

    in Fusarium commune. Prediction of the cellulose and hemicellulose-degrading enzymes in the F. commune transcriptome using peptide pattern recognition revealed 147 genes encoding glycoside hydrolases and six genes encoding lytic polysaccharide monooxygenases (AA9 and AA11), including all relevant cellulose......-d-xylanase and β-xylosidase activities; and XYL11 was a true xylanase characterized by high substrate specificity. These results indicate that F. commune with genetic modification is a promising source of enzymes for the decomposition of lignocellulosic biomass....

  2. Fungal cellulase/xylanase production and corresponding hydrolysis using pretreated corn stover as substrates.

    Science.gov (United States)

    Zhang, Liang; Wang, Xiaoqing; Ruan, Zhenhua; Liu, Ying; Niu, Xiaorui; Yue, Zhengbo; Li, Zhimin; Liao, Wei; Liu, Yan

    2014-01-01

    Three pretreated corn stover (ammonia fiber expansion, dilute acid, and dilute alkali) were used as carbon source to culture Trichoderma reesei Rut C-30 for cellulase and xylanase production. The results indicated that the cultures on ammonia fiber expansion and alkali pretreated corn stover had better enzyme production than the acid pretreated ones. The consequent enzymatic hydrolysis was performed applying fungal enzymes on pretreated corn stover samples. Tukey's statistical comparisons exhibited that there were significant differences on enzymatic hydrolysis among different combination of fungal enzymes and pretreated corn stover. The higher sugar yields were achieved by the enzymatic hydrolysis of dilute alkali pretreated corn stover.

  3. Production of beta-xylanase and beta-xylosidase by the extremely halophilic archaeon Halorhabdus utahensis

    DEFF Research Database (Denmark)

    Wainø, M.; Ingvorsen, K.

    2003-01-01

    The extremely halophilic archaeon, Halorhabdus utahensis, isolated from the Great Salt Lake, Utah, produced beta-xylanase and beta-xylosidase activities. Both enzymes were active over a broad NaCl range from near zero to 30% NaCl when tested with culture broth. A broad NaCl optimum was observed......-xylosidase activity was optimal at 65degreesC. SDS-PAGE and zymogram techniques revealed the presence of two xylan-degrading proteins of approximately 45 and 67 kDa in culture supernatants. To our knowledge, this paper is the first report on hemicellulose-degrading enzymes produced by an extremely halophilic archaeon....

  4. Xylanase Production from Trichoderma harzianum 1073 D3 with Alternative Carbon and Nitrogen Sources

    OpenAIRE

    Seyis,Isil; Aksoz, Nilufer

    2005-01-01

    The effect of some natural wastes (orange pomace, orange peel, lemon pomace, lemon peel, apple pomace, pear peel, banana peel, melon peel and hazelnut shell) on the production of xylanase from Trichoderma harzianum 1073 D3 has been studied and maximum activity has been observed on melon peel (26.5 U/mg of protein) followed by apple pomace and hazelnut shell. Also, molasses could be used as an additional carbon source as it decreased the production time approximately by 50 %. Finally, potentia...

  5. French vineyards provide information that opens ways for effective resistance management of Botrytis cinerea (grey mould).

    Science.gov (United States)

    Walker, Anne-Sophie; Micoud, Annie; Rémuson, Florent; Grosman, Jacques; Gredt, Michel; Leroux, Pierre

    2013-06-01

    Resistance to fungicides is an evolutionary process resulting from the selection of advantageous genotypes in naturally diverse populations. Seven fungicide modes of action are authorised to control grey mould caused by Botrytis cinerea on grapevine in France, and five of them have encountered specific resistance, with variable frequencies in populations and possible consequences for field fungicide efficacy. Moreover, multidrug resistance is caused by fungicide efflux and allows a weak resistance towards six unrelated modes of action. Here, a review is given of the fungicide resistance status of B. cinerea in France, particularly in the vineyards of Champagne, which are the most affected. Recently developed resistance and recent findings concerning the associated resistance mechanisms are focused upon in particular. Finally, antiresistance strategies are presented, and examples of managed resistance are discussed in a more general manner with the aim of extending this knowledge to other crops and countries undergoing similar resistance problems.

  6. Phytoconstituents and in vitro Evaluation of Antioxidant Capacities of Cotula Cinerea (Morocco Methanol Extracts

    Directory of Open Access Journals (Sweden)

    Farid Khallouki

    2015-06-01

    Full Text Available T he purpose of this study was to determine the phytochemical content of Cotula cinerea to establish principal components which may consolidate its use as a medicinal plant in the southeast of Morocco. The amount of total phenolic compounds as determined by analytical HPLC in methanol extracts was 79.23 ± 2.5 mg/g dry matter. The major phenolic compounds identified by HPLC-ESI-MS were neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, 3,4-dicaffeoylquinic acid, 3,5-dicaffeoylquinic acid, 4,5-dicaffeoylquinic acid and luteolin-4´-O-glucoside. All compounds displayed very strong antioxidant capacities in the DPPH, FRAP and ORAC assays . The data indicates that methanol extracts of C. cinerea via their antioxidant capacities, may be effective disease prevention potions in traditional African medicine which is probably related to the significant content of echinoids and flavonoids.

  7. Chemical Characterization of Different Sumac and Pomegranate Extracts Effective against Botrytis cinerea Rots

    OpenAIRE

    Romeo, Flora V.; Gabriele Ballistreri; Simona Fabroni; Sonia Pangallo; Maria Giulia Li Destri Nicosia; Leonardo Schena; Paolo Rapisarda

    2015-01-01

    Pomegranate (Punica granatum L.) peel and sumac (Rhus coriaria L.) fruit and leaf extracts were chemically characterized and their ability to inhibit table grape (cv. Italia) rots caused by Botrytis cinerea was evaluated on artificially inoculated berries. Different extraction methods were applied and extracts were characterized through Ultra Fast High Performance Liquid Chromatography coupled to Photodiode array detector and Electrospray ionization Mass spectrometer (UPLC-PDA-ESI/MSn) for th...

  8. Bombus terrestris as an entomovector for suppressing Botrytis cinerea in open field strawberry

    OpenAIRE

    Mänd, Marika; Karise, Reet; Muljar, Riin

    2013-01-01

    Strawberry (Fragaria x ananassa) is a fruit crop grown worldwide, but diseases such as the grey mould Botrytis cinerea frequently limit its yield. Most of grey mould infection on the fruits is initiated during the flowering period. Use of foraging bees as disseminators of microbial control agents (MCAs) to flowers is known as entomovector technology. Many researchers have shown that bumble bees can efficiently vector MCAs; however, most studies have been conducted in greenhouse conditions.

  9. The pathogenicity of different Botrytis cinerea Pers. isolates to apples and their sensitivity to benzimidazole fungicides

    OpenAIRE

    Hanna Bryk

    2013-01-01

    The pathogenicity of 80 isolates of Botrytis cinerea Pers. from different hosts to apple fruit was examined. Host specificity among isolates was not found. All of the isolates, independent of their derivation, caused apple fruit rot. Isolates from apple fruits showed moderate and strong pathogenicity to apple fruits. Only 1 of the 22 examined isolates showed weak pathogenicity. Tolerance to benomyl was compared among isolates obtained from apple fruits and from other hosts. It was found that ...

  10. Proctitis associated with Neisseria cinerea misidentified as Neisseria gonorrhoeae in a child.

    OpenAIRE

    Dossett, J H; Appelbaum, P. C.; Knapp, J S; Totten, P A

    1985-01-01

    An 8-year-old boy developed proctitis. Rectal swabs yielded a Neisseria sp. that was repeatedly identified by API (Analytab Products, Plainview, N.Y.), Minitek (BBL Microbiology Systems, Cockeysville, Md.), and Bactec (Johnston Laboratories, Towson, Md.) methods as Neisseria gonorrhoeae. Subsequent testing in a reference laboratory yielded an identification of Neisseria cinerea. It is suggested that identification of a Neisseria sp. isolated from genital or rectal sites in a child be confirme...

  11. Nosocomial pneumonia caused by a glucose-metabolizing strain of Neisseria cinerea.

    OpenAIRE

    Boyce, J M; Taylor, M R; Mitchell, E B; Knapp, J S

    1985-01-01

    We describe what appears to be the first reported case of nosocomial pneumonia caused by Neisseria cinerea. The isolate metabolized glucose when tested in BACTEC Neisseria Differentiation Kits (Johnston Laboratories), but did not produce detectable acid in cystine-Trypticase (BBL Microbiology Systems) agar medium or in modified oxidation-fermentation medium. Clinical laboratories that rely on the BACTEC method for differentiation of pathogenic neisseriae should be aware of the fact that N. ci...

  12. Biological characteristics and resistance analysis of the novel fungicide SYP-1620 against Botrytis cinerea.

    Science.gov (United States)

    Zhang, Xiaoke; Wu, Dongxia; Duan, Yabing; Ge, Changyan; Wang, Jianxin; Zhou, Mingguo; Chen, Changjun

    2014-09-01

    SYP-1620, a quinone-outside-inhibitor (QoI), is a novel broad-spectrum fungicide. In this study, 108 isolates of Botrytis cinerea from different geographical regions in Jiangsu Province of China were characterized for baseline sensitivity to SYP-1620. The curves of baseline sensitivity were unimodal with a mean EC50 value of 0.0130±0.0109 μg/mL for mycelial growth, 0.01147±0.0062 μg/mL for spore germination, respectively. The biological characterization of SYP-1620 against B. cinerea was determined in vitro. The results indicated that SYP-1620 has a strong inhibiting effect on spore germination, mycelial growth, and respiration. The protective and curative test of SYP-1620 suggested that protective effect was better than curative either on strawberry leaves or on cucumber leaves in vivo. In addition, the biological characterization of SYP-1620-resistant mutants of B. cinerea was investigated. SYP-1620 has no cross-resistance with other types of fungicide. Compared to the sensitive isolates, the resistant mutants had lower mycelial growth and virulence but not differ in mycelial dry weight. Sequencing indicated that SYP-1620 resistance was associated with a single point mutation (G143A) in the cytochrome b gene.

  13. Diversity of MHC DQB and DRB Genes in the Endangered Australian Sea Lion (Neophoca cinerea).

    Science.gov (United States)

    Lau, Quintin; Chow, Natalie; Gray, Rachael; Gongora, Jaime; Higgins, Damien P

    2015-01-01

    Major histocompatibility complex (MHC) class II molecules have an important role in vertebrate adaptive immunity, being responsible for recognizing, binding, and presenting specific antigenic peptides to T lymphocytes. Here, we study the MHC class II DQB and DRB exon 2 genes of the Australian sea lion (Neophoca cinerea), an endangered pinniped species that experiences high pup mortality. Following characterization of N. cinerea DQB and DRB by molecular cloning, and evaluation of diversity in pups across 2 colonies using variant screening (n = 47), 3 DQB alleles and 10 DRB variants (including 1 pseudogene allele) were identified. The higher diversity at DRB relative to DQB is consistent with other studies in marine mammals. Despite overall lower MHC class II allelic diversity relative to some other pinniped species, we observed similar levels of nucleotide diversity and selection in N. cinerea. In addition, we provide support for recent divergence of MHC class II alleles. The characterization of MHC class II diversity in the Australian sea lion establishes a baseline for further investigation of associations with disease, including endemic hookworm infection, and contributes to the conservation management of this species.

  14. Development of Botrytis cinerea Pers. ex Fr. on leaves of common poinsettia (Euphorbia pulcherrima Willd.

    Directory of Open Access Journals (Sweden)

    Beata Kułek

    2012-12-01

    Full Text Available The development of Botrytis cinerea was assessed on six cultivars of common poinsettia, differing in the colour of bracts, and being in great demand among buyers of these ornamental plants. Resistance to this pathogen differed in the investigated poinsettias. Cultivar 'Malibu Red' (red bracts turned out to be most susceptible, while cv. 'Marblestar' (cream-pink and cv. 'Coco White' (white - relatively resistant to this fungus. After application of various inoculation methods (leaf discs, cut off leaves, whole plants the differences in resistance to B. cinerea were confirmed for two extreme cultivars - susceptible ('Malibu Red' and resistant ('Coco White', which indicated genetic background of this polymorphism. The rate of disease development on poinsettia leaves was affected by the amount of spores used for inoculation (optimum density of 3.5·105 B. cinerea conidia / ml suspension and the addition of stimulants (0.1 M glucose with 0.05 M KH2PO4, which facilitated germination and infection of the host tissue. The inoculated poinsettia leaves showed high stability of plasma membranes. In the susceptible cultivar, in spite of the development of necrotic spots, a significant increase in the membrane damage index (by 13% was found only on day 7 of the disease development.

  15. Host-targeting-motif Harbored Secretary Proteins in Genome of Plant Pathogenic Fungus Botrytis cinerea

    Institute of Scientific and Technical Information of China (English)

    Zhang Yue; Chen Zi-niu; Su Yuan; Yu Lei

    2012-01-01

    According to our previous study, saprophytic fungi Botrytis cinerea contained 579 predicted secretary proteins. Among them, we found that 122 of these proteins contained the highly conserved pathogenic-related host-targeting-motif RxLx within 100 residues adjacent to the signal peptide cleavage site. According to PEDNAT and COG of the GenBank database, the functions of this motif containing proteins included metabolism modification and cell secretion. We blasted them in GenBank and found 47.54% had highly conserved homologues in other species, among them 74.1% had putative functional domains. This suggests these proteins are presumably ancient and vertically transmitted within the species. Many of these domains belonged to proteins which played roles in the pathogenic process of other kinds of pathogens and some had already been proved to be pathogenic secretary proteins of Botrytis cinerea. So we postulated that proteins contained host-targeting-motif RxLx were candidates participating in the pathogenesis of Botrytis cinerea.

  16. Interaction of Ulocladium atrum, a Potential Biological Control Agent, with Botrytis cinerea and Grapevine Plantlets

    Directory of Open Access Journals (Sweden)

    Sébastien Ronseaux

    2013-09-01

    Full Text Available The effectiveness of biological control agent, Ulocladium atrum (isolates U13 and U16 in protecting Vitis vinifera L. cv. Chardonnay against gray mold disease caused by Botrytis cinerea, and simulation of the foliar defense responses was investigated. A degraded mycelium structure during cultural assay on potato dextrose agar revealed that U. atrum isolates U13 and U16 were both antagonistic to B. cinerea, mainly when isolates were inoculated two days before Botrytis. Under in vitro conditions, foliar application of U. atrum protected grapevine leaves against gray mold disease. An increase in chitinase activity was induced by the presence of U. atrum isolates indicating that the biological control agents triggered plant defense mechanisms. Moreover, U13 has the potential to colonize the grapevine plantlets and to improve their growth. The ability of U. atrum isolates to exhibit an antagonistic effect against B. cinerea in addition to their aptitude to induce plant resistance and to promote grapevine growth may explain a part of their biological activity. Hence, this study suggests that U. atrum provides a suitable biocontrol agent against gray mold in grapevines.

  17. Laccase production by free and immobilized mycelia of Peniophora cinerea and Trametes versicolor: a comparative study.

    Science.gov (United States)

    Silvério, Sara C; Moreira, Sérgio; Milagres, Adriane M F; Macedo, Eugénia A; Teixeira, José A; Mussatto, Solange I

    2013-03-01

    The production of laccase by immobilized mycelia of Peniophora cinerea and Trametes versicolor was studied. In an initial stage, experimental assays were performed in Erlenmeyer flasks using free and immobilized mycelium, and the performance of the fungal strains to produce the enzyme was compared. Both fungi adhered into the support material (a synthetic fiber), growing not only on the surface but also in the interspaces of the fibers. Immobilization of P. cinerea provided a 35-fold increase in laccase production when compared to the production obtained by using free mycelium. On the other hand, immobilization of T. versicolor caused a decrease in laccase activity. A comparison between the strains revealed that immobilized P. cinerea (3,500 U/L) surpassed the enzyme production by free T. versicolor (800 U/L). When the conditions that gave the best laccase production to each fungus were employed in a stirred tank bioreactor, very low laccase production was observed for both the cases, suggesting that shear stress and mycelia damage caused by the agitation impellers negatively affected the enzyme production.

  18. RNAseq-based transcriptome analysis of Lactuca sativa infected by the fungal necrotroph Botrytis cinerea.

    Science.gov (United States)

    De Cremer, Kaat; Mathys, Janick; Vos, Christine; Froenicke, Lutz; Michelmore, Richard W; Cammue, Bruno P A; De Coninck, Barbara

    2013-11-01

    The fungal pathogen Botrytis cinerea establishes a necrotrophic interaction with its host plants, including lettuce (Lactuca sativa), causing it to wilt, collapse and eventually dry up and die, which results in serious economic losses. Global expression profiling using RNAseq and the newly sequenced lettuce genome identified a complex network of genes involved in the lettuce-B. cinerea interaction. The observed high number of differentially expressed genes allowed us to classify them according to the biological pathways in which they are implicated, generating a holistic picture. Most pronounced were the induction of the phenylpropanoid pathway and terpenoid biosynthesis, whereas photosynthesis was globally down-regulated at 48 h post-inoculation. Large-scale comparison with data available on the interaction of B. cinerea with the model plant Arabidopsis thaliana revealed both general and species-specific responses to infection with this pathogen. Surprisingly, expression analysis of selected genes could not detect significant systemic transcriptional alterations in lettuce leaves distant from the inoculation site. Additionally, we assessed the response of these lettuce genes to a biotrophic pathogen, Bremia lactucae, revealing that similar pathways are induced during compatible interactions of lettuce with necrotrophic and biotrophic pathogens.

  19. Botrytis cinerea protein O-mannosyltransferases play critical roles in morphogenesis, growth, and virulence.

    Directory of Open Access Journals (Sweden)

    Mario González

    Full Text Available Protein O-glycosylation is crucial in determining the structure and function of numerous secreted and membrane-bound proteins. In fungi, this process begins with the addition of a mannose residue by protein O-mannosyltransferases (PMTs in the lumen side of the ER membrane. We have generated mutants of the three Botrytis cinerea pmt genes to study their role in the virulence of this wide-range plant pathogen. B. cinerea PMTs, especially PMT2, are critical for the stability of the cell wall and are necessary for sporulation and for the generation of the extracellular matrix. PMTs are also individually required for full virulence in a variety of hosts, with a special role in the penetration of intact plant leaves. The most significant case is that of grapevine leaves, whose penetration requires the three functional PMTs. Furthermore, PMT2 also contributes significantly to fungal adherence on grapevine and tobacco leaves. Analysis of extracellular and membrane proteins showed significant changes in the pattern of protein secretion and glycosylation by the pmt mutants, and allowed the identification of new protein substrates putatively glycosylated by specific PMTs. Since plants do no possess these enzymes, PMTs constitute a promising target in the development of novel control strategies against B. cinerea.

  20. Antifungal activity of resveratrol against Botrytis cinerea is improved using 2-furyl derivatives.

    Directory of Open Access Journals (Sweden)

    Francesco Caruso

    Full Text Available The antifungal effect of three furyl compounds closely related to resveratrol, (E-3,4,5-trimethoxy-β-(2-furyl-styrene (1, (E-4-methoxy-β-(2-furyl-styrene (2 and (E-3,5-dimethoxy-β-(2-furyl-styrene (3 against Botrytis cinerea was analyzed. The inhibitory effect, at 100 µg ml(-1 of compounds 1, 2, 3 and resveratrol on conidia germination, was determined to be about 70%, while at the same concentration pterostilbene (a dimethoxyl derivative of resveratrol produced complete inhibition. The title compounds were more fungitoxic towards in vitro mycelial growth than resveratrol and pterostilbene. Compound 3 was the most active and a potential explanation of this feature is given using density functional theory (DFT calculations on the demethoxylation/demethylation process. Compound 3 was further evaluated for its effects on laccase production, oxygen consumption and membrane integrity of B. cinerea. An increase of the laccase activity was observed in the presence of compound 3 and, using Sytox Green nucleic acid stain, it was demonstrated that this compound altered B. cinerea membrane. Finally, compound 3 partially affected conidia respiration.

  1. Effect of additives on adsorption and desorption behavior of xylanase on acid-insoluble lignin from corn stover and wheat straw.

    Science.gov (United States)

    Li, Yanfei; Ge, Xiaoyan; Sun, Zongping; Zhang, Junhua

    2015-06-01

    The competitive adsorption between cellulases and additives on lignin in the hydrolysis of lignocelluloses has been confirmed, whereas the effect of additives on the interaction between xylanase and lignin is not clear. In this work, the effects of additives, poly(ethylene glycol) 2000, poly(ethylene glycol) 6000, Tween 20, and Tween 80, on the xylanase adsorption/desorption onto/from acid-insoluble lignin from corn stover (CS-lignin) and wheat straw (WS-lignin) were investigated. The results indicated that the additives could adsorb onto isolated lignin and reduce the xylanase adsorption onto lignin. Compared to CS-lignin, more additives could adsorb onto WS-lignin, making less xylanase adsorbed onto WS-lignin. In addition, the additives could enhance desorption of xylanase from lignin, which might be due to the competitive adsorption between xylanase and additives on lignin. The released xylanase from lignin still exhibited hydrolytic capacity in the hydrolysis of isolated xylan and xylan in corn stover.

  2. Optimization of Xylanase Production through Response Surface Methodology by Fusarium sp. BVKT R2 Isolated from forest soil and its applications in saccharification

    Directory of Open Access Journals (Sweden)

    Ramanjaneyulu Golla

    2016-09-01

    Full Text Available AbstractXylanses are hydrolytic enzymes with wide applications in several industries like biofuels, paper and pulp, deinking, food and feed. The present study was aimed at hitting at high yield xylanase producing fungi from natural resources. Two highest xylanase producing fungal isolates - Q12 and L1were picked from collection of 450 fungal cultures for the utilization of xylan. These fungal isolates - Q12 and L1 were identified basing on ITS gene sequencing analysis as Fusarium sp. BVKT R2 (KT119615 and Fusarium strain BRR R6 (KT119619, respectively with construction of phylogenetic trees. Fusarium sp. BVKT R2 was further optimized for maximum xylanase production and the interaction effects between variables on production of xylanase were studied through response surface methodology. The optimal conditions for maximal production of xylanase were sorbitol 1.5%, yeast extract 1.5%, pH of 5.0, Temperature of 32.5ºC, and agitation of 175 rpm. Under optimal conditions, the yields of xylanase production by Fusarium sp. BVKT R2 was as high as 4560 U/ml in SmF. Incubation of different lignocellulosic biomasses with crude enzyme of Fusarium sp. BVKT R2 at 37°C for 72 h could achieve about 45% saccharification. The results suggest that Fusarium sp. BVKT R2 has potential applications in saccharification process of biomass.Key words: Fusarium sp., Optimization, Response Surface Methodology, Saccharification, Submerged fermentation, Xylanase

  3. Strategies of xylanase supplementation for an efficient saccharification and cofermentation process from pretreated wheat straw.

    Science.gov (United States)

    Alvira, Pablo; Tomás-Pejó, Elia; Negro, María José; Ballesteros, Mercedes

    2011-07-01

    Ethanol production from lignocellulosic raw materials includes a pretreatment step before enzymatic hydrolysis (EH). Pretreated substrates contain complex hemicelluloses in the solid fraction that can protect the cellulose from enzymatic attack. In addition, soluble xylooligomers are contained in the pretreated materials and may have an inhibitory effect on cellulase activity. In this context, several approaches for xylanase supplementation have been studied to increase EH yields. In this study, the whole slurry obtained after steam explosion pretreatment of wheat straw has been used as substrate. EH experiments were performed using commercial cellulase preparations supplemented with an endoxylanase (XlnC) from Aspergillus nidulans. Among different strategies of XlnC supplementation, the 24-h xylanase treatment before cellulase addition yielded an increase of 40.1 and 10.1% in glucose and xylose production, respectively. Different XlnC addition strategies were integrated in a simultaneous saccharification and cofermentation process (SSCF) using the xylose fermenting strain Saccharomyces cerevisiae F12. Ethanol production in SSCF was 28.4% higher when comparing to a simultaneous saccharification and fermentation process.

  4. Contribution of ethanol-tolerant xylanase G2 from Aspergillus oryzae on Japanese sake brewing.

    Science.gov (United States)

    Sato, Yuichiro; Fukuda, Hisashi; Zhou, Yan; Mikami, Shigeaki

    2010-12-01

    We purified three xylanase isozymes (XynF1, XynF3 and XynG2) from a solid-state Aspergillus oryzae RIB128 culture using chromatography. The results of our sake-brewing experiment, in which we used exogenously supplemented enzymes, revealed that only XynG2 improved the alcohol yield and the material utilization. The alcohol yield of the XynG2 batch displayed an increase of 4.4% in comparison to the control, and the amount of sake cake decreased by 4.6%. The contribution of XynG2 was further confirmed through our brewing experiment in which we used the yeast heterogeneously expressing fungal xylanase isozymes. Interestingly XynG1, an enzyme with a XynG2-like sequence that is more vulnerable to ethanol, did not improve the sake-mash fermentation. The stability of XynG2 in ethanol was prominent, and it retained most of its original activity after we exposed it to 80% ethanol for 30min, whereas the stability of the other isozymes in ethanol, including XynG1, was much lower (20-25% ethanol). We concluded, therefore, that the improvement of material utilization achieved with XynG2 is primarily attributable to its characteristically high stability in ethanol, thereby, effectively degrading rice endosperm cell walls under high-alcohol conditions such as a sake-mash environment.

  5. Structural Analysis of Xylanase from Marine Thermophilic Geobacillus stearothermophilus in Tanjung Api, Poso, Indonesia

    Directory of Open Access Journals (Sweden)

    BUDI SAKSONO

    2010-12-01

    Full Text Available A xylanase gene, xynA, has been cloned from thermophilic strain Geobacillus stearothermophilus, which was isolated from marine Tanjung Api, Indonesia. The polymerase chain reaction product of 1266 bp of xynA gene consisted of 1221 bp open reading frame and encoded 407 amino acids including 30 residues of signal peptide. The sequence exhibited highest identity of 98.7% in the level of amino acid, with an extracellular endo-1,4-β-xylanase from G. stearothermophilus T-6 (E-GSX T-6 of the glycoside hydrolase family 10 (GH10. A comparative study between the local strain G. stearothermophilus (GSX L and E-GSX T-6 on homology of amino acid sequence indicated five differents amino acids in the gene. They were Threonine/Alanine (T/A, Asparagine/Aspartate (N/D, Lysine/Asparagine (K/N, Isoleucine/Methionine (I/M, Serine/Threonine (S/T at the position 220, 227, 228, 233, and 245, respectively. Protein structural analysis of those differences suggested that those amino acids may play role in biochemical properties such as enzyme stability, in particular its thermostability.

  6. Endo-xylanase and endo-cellulase-assisted extraction of pectin from apple pomace.

    Science.gov (United States)

    Wikiera, Agnieszka; Mika, Magdalena; Starzyńska-Janiszewska, Anna; Stodolak, Bożena

    2016-05-20

    Pectins were extracted from apple pomace with monoactive preparation of endo-xylanase and endo-cellulase. The process was conducted for 10 h in conditions of pH 5.0 at 40 °C, with constant shaking. Endo-xylanase application resulted in the highest extraction efficiency of pectins (19.8%). The obtained polymer was characterised by a very high molecular mass, high level of neutral sugars - mainly arabinose, galactose and glucose, and very high DM (73.4). It also contained the highest level of protein and phenols. Pectin extracted with endo-cellulase had 1.5 fold lower molecular mass but contained significantly more GalA (70.5%) of a high degree of methylation (66.3%). The simultaneous application of both enzymatic preparations resulted in their cooperation, leading to a decrease of both the extraction efficiency and the molecular mass of pectin. However, this pectin was distinguished by the highest GalA (74.7%) and rhamnose contents.

  7. Cellulase and xylanase activity during the decomposition of three aquatic macrophytes in a tropical oxbow lagoon

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    L Sciessere

    2011-09-01

    Full Text Available Due to the connection between enzymatic activity and degradation of different fractions of organic matter, enzyme assays can be used to estimate degradation rates of particulate and dissolved organic carbon in freshwater systems. The aim of this study was to quantify and model the enzymatic degradation involving the decomposition of macrophytes, describing temporal activity of cellulases (EC 3.2.1.4 and EC 3.2.1.91 and xylanase (EC 3.2.1.8 during in situ decomposition of three aquatic macrophytes (Salvinia sp., Eichhornia azurea and Cyperus giganteus on the surface and water-sediment interface (w-s interface of an oxbow lagoon (Óleo lagoon within a natural Brazilian Savanna Reserve. Overall, the enzymatic degradation of aquatic macrophytes in Óleo lagoon occurred during the whole year and was initiated together with leaching. Xylanase production was ca. 5 times higher than cellulase values due to easy access to this compound by cellulolytic microorganisms. Enzymatic production and detritus mass decay were similar on the surface and w-s interface. Salvinia sp. was the most recalcitrant detritus, with low mass decay and enzymatic activity. E. azurea and C. giganteus decomposition rates and enzymatic production were high and similar. Due to the physicochemical homogeneity observed in the Óleo lagoon, the differences between the decay rates of each species are mostly related with detritus chemical quality.

  8. A thermostable Gloeophyllum trabeum xylanase with potential for the brewing industry.

    Science.gov (United States)

    Wang, Xiaoyu; Luo, Huiying; Yu, Wangning; Ma, Rui; You, Shuai; Liu, Weina; Hou, Lingyu; Zheng, Fei; Xie, Xiangming; Yao, Bin

    2016-05-15

    A xylanase gene of glycoside hydrolase family 10, GtXyn10, was cloned from Gloeophyllum trabeum CBS 900.73 and expressed in Pichia pastoris GS115. Purified recombinant GtXyn10 exhibited significant activities to xylan (100.0%), lichenan (11.2%), glucan (15.2%) and p-nitrophenol-β-cellobiose (18.6%), demonstrated the maximum xylanase and glucanase activities at pH 4.5-5.0 and 75°C, retained stability over the pH range of 2.0-7.5 and at 70°C, and was resistant to pepsin and trypsin, most metal ions and SDS. Multiple sequence alignment and modeled-structure analysis identified a unique Gly48 in GtXyn10, and site-directed mutagenesis of Gly48 to Lys improved the temperature optimum up to 80°C. Under simulated mashing conditions, GtXyn10 (80U) reduced the mash viscosity by 12.8% and improved the filtration rate by 31.3%. All these properties above make GtXyn10 attractive for potential applications in the feed and brewing industries.

  9. Cellulase and Xylanase Production by Penicillium echinulatum in Submerged Media Containing Cellulose Amended with Sorbitol

    Directory of Open Access Journals (Sweden)

    Carla Eliana Todero Ritter

    2013-01-01

    Full Text Available The present work investigated the use of sorbitol as a soluble carbon source, in association with cellulose, to produce cellulases and xylanases in submerged cultures of Penicillium echinulatum 9A02S1. Because cellulose is an insoluble carbon source, in cellulase production, there are some problems with rheology and oxygen transfer. The submerged fermentations containing media composed of 0, 0.25, 0.5, 0.75, and 1% (w/v sorbitol and cellulose that were added at different times during the cultivation; 0.2% (w/v soy bran; 0.1% (w/v wheat bran; and a solution of salts. The highest filter paper activity (FPA ( IU·mL−1 was obtained on the seventh day in the medium containing 0.5% (w/v sorbitol and 0.5% (w/v cellulose added 24 h after the start of cultivation. However, the CMCases showed an activity peak on the sixth day ( IU·mL−1 in the medium containing 0.75% (w/v sorbitol and 0.75% (w/v cellulose added after 12 h of cultivation. The xylanases showed the highest activity in the medium with 0.75% (w/v sorbitol and 0.25% (w/v cellulose added 36 h after the start of cultivation. This strategy enables the reduction of the cellulose concentration, which in high concentrations can cause rheological and oxygen transfer problems.

  10. Dynamics in the Strawberry Rhizosphere Microbiome in Response to Biochar and Botrytis cinerea Leaf Infection

    Science.gov (United States)

    De Tender, Caroline; Haegeman, Annelies; Vandecasteele, Bart; Clement, Lieven; Cremelie, Pieter; Dawyndt, Peter; Maes, Martine; Debode, Jane

    2016-01-01

    Adding biochar, the solid coproduct of biofuel production, to peat can enhance strawberry growth, and disease resistance against the airborne fungal pathogen Botrytis cinerea. Additionally, biochar can induce shifts in the strawberry rhizosphere microbiome. However, the moment that this biochar-mediated shift occurs in the rhizosphere is not known. Further, the effect of an above-ground infection on the strawberry rhizosphere microbiome is unknown. In the present study we established two experiments in which strawberry transplants (cv. Elsanta) were planted either in peat or in peat amended with 3% biochar. First, we established a time course experiment to measure the effect of biochar on the rhizosphere bacterial and fungal communities over time. In a second experiment, we inoculated the strawberry leaves with B. cinerea, and studied the impact of the infection on the rhizosphere bacterial community. The fungal rhizosphere community was stabilized after 1 week, except for the upcoming Auriculariales, whereas the bacterial community shifted till 6 weeks. An effect of the addition of biochar to the peat on the rhizosphere microbiome was solely measured for the bacterial community from week 6 of plant growth onwards. When scoring the plant development, biochar addition was associated with enhanced root formation, fruit production, and postharvest resistance of the fruits against B. cinerea. We hypothesize that the bacterial rhizosphere microbiome, but also biochar-mediated changes in chemical substrate composition could be involved in these events. Infection of the strawberry leaves with B. cinerea induced shifts in the bacterial rhizosphere community, with an increased bacterial richness. This disease-induced effect was not observed in the rhizospheres of the B. cinerea-infected plants grown in the biochar-amended peat. The results show that an above-ground infection has its effect on the strawberry rhizosphere microbiome, changing the bacterial interactions in the

  11. Constitutive expression of the xylanase inhibitor TAXI-III delays Fusarium head blight symptoms in durum wheat transgenic plants.

    Science.gov (United States)

    Moscetti, Ilaria; Tundo, Silvio; Janni, Michela; Sella, Luca; Gazzetti, Katia; Tauzin, Alexandra; Giardina, Thierry; Masci, Stefania; Favaron, Francesco; D'Ovidio, Renato

    2013-12-01

    Cereals contain xylanase inhibitor (XI) proteins which inhibit microbial xylanases and are considered part of the defense mechanisms to counteract microbial pathogens. Nevertheless, in planta evidence for this role has not been reported yet. Therefore, we produced a number of transgenic plants constitutively overexpressing TAXI-III, a member of the TAXI type XI that is induced by pathogen infection. Results showed that TAXI-III endows the transgenic wheat with new inhibition capacities. We also showed that TAXI-III is correctly secreted into the apoplast and possesses the expected inhibition parameters against microbial xylanases. The new inhibition properties of the transgenic plants correlate with a significant delay of Fusarium head blight disease symptoms caused by Fusarium graminearum but do not significantly influence leaf spot symptoms caused by Bipolaris sorokiniana. We showed that this contrasting result can be due to the different capacity of TAXI-III to inhibit the xylanase activity of these two fungal pathogens. These results provide, for the first time, clear evidence in planta that XI are involved in plant defense against fungal pathogens and show the potential to manipulate TAXI-III accumulation to improve wheat resistance against F. graminearum.

  12. Crystallization and preliminary crystallographic analysis of endo-1,4-beta-xylanase I from Aspergillus niger

    NARCIS (Netherlands)

    Krengel, U; Rozeboom, HJ; Kalk, KH; Dijkstra, BW

    1996-01-01

    A family G xylanase from Aspergillus niger has been crystallized using the vapor-diffusion method. Several crystal forms could be obtained using various sodium salts as precipitants. Three of the crystal forms belong to space groups P2(1), P2(1)2(1)2(1) and P4(3) and have cell parameters of approxim

  13. Agar-agar entrapment increases the stability of endo-β-1,4-xylanase for repeated biodegradation of xylan.

    Science.gov (United States)

    Bibi, Zainab; Shahid, Faiza; Ul Qader, Shah Ali; Aman, Afsheen

    2015-04-01

    Microbial xylanases, specially endo-β-1,4-xylanase catalyzes the hydrolysis of xylan, is considered one of the most significant hydrolases. It has numerous applications but most extensively is utilized in paper and pulp industry as a bio-bleaching agent. Immobilization technique is comprehensively studied with the expectation of modifying and improving enzyme stability and characteristics for commercial purposes. Currently, matrix entrapment technique is applied to immobilize endo-β-1,4-xylanase within agar-agar gel beads produced by Geobacillus stearothermophilus KIBGE-IB29. Maximal enzyme immobilization yield was achieved at 2.5% of agar-agar concentration. Optimized conditions demonstrated an increase in the optimal reaction time from 05 min to 30 min and incubation temperature from 50 °C to 60 °C with reference to free enzyme whereas; no effect was observed for optimum pH. Entrapment technique uniquely changed the kinetic parameters of immobilized endo-β-1,4-xylanase (Km: 0.5074 mg min(-1) to 0.5230 mg min(-1) and Vmax: 4773 U min(-1) to 968 U min(-1)) as compared to free enzyme. However, immobilized enzyme displayed broad thermal stability and retained 79.0% of its initial activity at 80 °C up to 30 min whereas; free enzyme completely lost its activity at this temperature. With respect to economic feasibility, the immobilized enzyme showed impressive recycling efficiency up to six reaction cycles.

  14. Fusion of carbohydrate binding modules from Thermotoga neapolitana with a family 10 xylanase from Bacillus halodurans S7.

    Science.gov (United States)

    Mamo, Gashaw; Hatti-Kaul, Rajni; Mattiasson, Bo

    2007-01-01

    Xylanase A of Thermotoga neapolitana contains binding domains both at the N- and C-terminal ends of the catalytic domain. In the N-terminal position it contains two carbohydrate-binding modules (CBM) which belong to family 22. These CBMs bind xylan but not to cellulose. The gene encoding the mature peptide of these CBMs was fused with an alkaline active GH10 xylanase from Bacillus halodurans S7 and expressed in Escherichia coli. The (His)(6) tagged hybrid protein was purified by immobilized metal affinity chromatography and characterized. Xylan binding by the chimeric protein was influenced by NaCl concentration and pH of the binding medium. Binding increased with increasing salt concentration up to 200 mM. Higher extent of binding was observed under acidic conditions. The fusion of the CBM structures enhanced the hydrolytic efficiency of the xylanase against insoluble xylan, but decreased the stability of the enzyme. The optimum temperature and pH for the activity of the xylanase did not change.

  15. Engineering increased thermostability in the GH-10 endo-1,4-ß-xylanase from Thermoascus aurantiacus CBMAI 756

    Science.gov (United States)

    The GH10 endo-xylanase from Thermoascus aurantiacus CBMAI 756 (XynA) is industrially attractive due to its considerable thermostability and high specific activity. Considering the possibility of a further improvement in thermostability, eleven mutants were created in the present study via site-direc...

  16. Seven N-terminal residues of a thermophilic xylanase are sufficient to confer hyperthermostability on its mesophilic counterpart.

    Directory of Open Access Journals (Sweden)

    Shan Zhang

    Full Text Available Xylanases, and especially thermostable xylanases, are increasingly of interest for the deconstruction of lignocellulosic biomass. In this paper, the termini of a pair of xylanases, mesophilic SoxB and thermophilic TfxA, were studied. Two regions in the N-terminus of TfxA were discovered to be potentially important for the thermostability. By focusing on Region 4, it was demonstrated that only two mutations, N32G and S33P cooperated to improve the thermostability of mesophilic SoxB. By introducing two potential regions into SoxB in combination, the most thermostable mutant, M2-N32G-S33P, was obtained. The M2-N32G-S33P had a melting temperature (Tm that was 25.6°C higher than the Tm of SoxB. Moreover, M2-N32G-S33P was even three-fold more stable than TfxA and had a Tm value that was 9°C higher than the Tm of TfxA. Thus, for the first time, the mesophilic SoxB "pupil" outperformed its thermophilic TfxA "master" and acquired hyperthermostability simply by introducing seven thermostabilizing residues from the extreme N-terminus of TfxA. This work suggested that mutations in the extreme N-terminus were sufficient for the mesophilic xylanase SoxB to acquire hyperthermostability.

  17. Cloning and in-silico analysis of beta-1,3-xylanase from psychrophilic yeast, Glaciozyma antarctica PI12

    Science.gov (United States)

    Nor, Nooraisyah Mohamad; Bakar, Farah Diba Abu; Mahadi, Nor Muhammad; Murad, Abdul Munir Abdul

    2015-09-01

    A beta-1,3-xylanase (EC 3.2.1.32) gene from psychrophilic yeast, Glaciozyma antarctica has been identified via genome data mining. The enzyme was grouped into GH26 family based on Carbohydrate Active Enzyme (CaZY) database. The molecular weight of this protein was predicted to be 42 kDa and is expected to be soluble for expression. The presence of signal peptide suggested that this enzyme may be released extracellularly into the marine environment of the host's habitat. This supports the theory that such enzymatic activity is required for degradation of nutrients of polysaccharide origins into simpler carbohydrates outside the environment before it could be taken up inside the cell. The sequence for this protein showed very little conservation (< 30%) with other beta-1,3-xylanases from available databases. Based on the phylogenetic analysis, this protein also showed distant relationship to other xylanases from eukaryotic origin. The protein may have undergone major substitution in its gene sequence order to adapt to the cold climate. This is the first report of beta-1,3-xylanase gene isolated from a psychrophilic yeast.

  18. Synthesis, processing and export of cytoplasmic endo-ß-1,4-xylanase from barley aleurone during germination

    NARCIS (Netherlands)

    Caspers, M.P.M.; Lok, F.; Sinjorgo, K.M.C.; Zeijl, M. van; Nielsen, K.A.; Cameron-Mills, V.

    2001-01-01

    We have identified the major endo-β-l,4-xylanase (XYN-1) in the aleurone of germinating barley grain, and show that it is expressed as a precursor of Mr 61 500 with both N- and C-terminal propeptides. XYN-1 is synthesized as an inactive enzyme in the cytoplasm, and only becomes active at a late stag

  19. Thermal-induced conformational changes in the product release area drive the enzymatic activity of xylanases 10B: Crystal structure, conformational stability and functional characterization of the xylanase 10B from Thermotoga petrophila RKU-1

    Energy Technology Data Exchange (ETDEWEB)

    Santos, Camila Ramos; Meza, Andreia Navarro [Laboratorio Nacional de Biociencias (LNBio), Centro Nacional de Pesquisa em Energia e Materiais, Campinas, SP (Brazil); Hoffmam, Zaira Bruna; Silva, Junio Cota; Alvarez, Thabata Maria; Ruller, Roberto [Laboratorio Nacional de Ciencia e Tecnologia do Bioetanol (CTBE), Centro Nacional de Pesquisa em Energia e Materiais, Campinas, SP (Brazil); Giesel, Guilherme Menegon; Verli, Hugo [Centro de Biotecnologia, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS (Brazil); Squina, Fabio Marcio [Laboratorio Nacional de Ciencia e Tecnologia do Bioetanol (CTBE), Centro Nacional de Pesquisa em Energia e Materiais, Campinas, SP (Brazil); Prade, Rolf Alexander [Department of Microbiology and Molecular Genetics, Oklahoma State University, Stillwater, OK (United States); Murakami, Mario Tyago, E-mail: mario.murakami@lnbio.org.br [Laboratorio Nacional de Biociencias (LNBio), Centro Nacional de Pesquisa em Energia e Materiais, Campinas, SP (Brazil)

    2010-12-10

    Research highlights: {yields} The hyperthermostable xylanase 10B from Thermotoga petrophila RKU-1 produces exclusively xylobiose at the optimum temperature. {yields} Circular dichroism spectroscopy suggests a coupling effect of temperature-induced structural changes with its enzymatic behavior. {yields} Crystallographic and molecular dynamics studies indicate that conformational changes in the product release area modulate the enzyme action mode. -- Abstract: Endo-xylanases play a key role in the depolymerization of xylan and recently, they have attracted much attention owing to their potential applications on biofuels and paper industries. In this work, we have investigated the molecular basis for the action mode of xylanases 10B at high temperatures using biochemical, biophysical and crystallographic methods. The crystal structure of xylanase 10B from hyperthermophilic bacterium Thermotoga petrophila RKU-1 (TpXyl10B) has been solved in the native state and in complex with xylobiose. The complex crystal structure showed a classical binding mode shared among other xylanases, which encompasses the -1 and -2 subsites. Interestingly, TpXyl10B displayed a temperature-dependent action mode producing xylobiose and xylotriose at 20 {sup o}C, and exclusively xylobiose at 90 {sup o}C as assessed by capillary zone electrophoresis. Moreover, circular dichroism spectroscopy suggested a coupling effect of temperature-induced structural changes with this particular enzymatic behavior. Molecular dynamics simulations supported the CD analysis suggesting that an open conformational state adopted by the catalytic loop (Trp297-Lys326) provokes significant modifications in the product release area (+1,+2 and +3 subsites), which drives the enzymatic activity to the specific release of xylobiose at high temperatures.

  20. Phylogenetic analysis of β-xylanase SRXL1 of Sporisorium reilianum and its relationship with families (GH10 and GH11) of Ascomycetes and Basidiomycetes

    OpenAIRE

    Jorge Álvarez-Cervantes; Gerardo Díaz-Godínez; Yuridia Mercado-Flores; Vijai Kumar Gupta; Miguel Angel Anducho-Reyes

    2016-01-01

    In this paper, the amino acid sequence of the β-xylanase SRXL1 of Sporisorium reilianum, which is a pathogenic fungus of maize was used as a model protein to find its phylogenetic relationship with other xylanases of Ascomycetes and Basidiomycetes and the information obtained allowed to establish a hypothesis of monophyly and of biological role. 84 amino acid sequences of β-xylanase obtained from the GenBank database was used. Groupings analysis of higher-level in the Pfam database allowed to...

  1. Control Effect and Possible Mechanism of the Natural Compound Phenazine-1-Carboxamide against Botrytis cinerea.

    Science.gov (United States)

    Zhang, Ya; Wang, Chong; Su, Pin; Liao, Xiaolan

    2015-01-01

    To develop new agents against strawberry grey mould and to aid in the development of biological pesticides, we investigated the inhibitory effect of a natural compound, phenazine-1-carboxamide (PCN), against Botrytis cinerea using a growth rate assay. Additionally, indoor toxicity and the in vitro control effect of PCN were further studied to determine its potential mechanisms of action on B. cinerea. PCN was inhibitory against B. cinerea with a 50% effective concentration (EC50) of 108.12 μg/mL; the toxicity of PCN was equivalent to that of carbendazim (CBM). The best in vitro control effect of PCN against grey mould in strawberry (fruit) reached 75.32%, which was slightly higher than that of CBM. The field control effect of PCN against grey mould reached a maximum of 72.31% at a PCN concentration of 700 μg/mL, which was 1.02 times higher than that of CBM. Fungistatic activity was observed at low concentrations of PCN, while high concentrations of PCN resulted in fungicidal activity against B. cinerea. This natural compound strongly inhibited both spore and sclerotium germination of B. cinerea, with the best relative inhibition rates of 77.03% and 82.11%, respectively. The inhibitory effect of PCN on mycelial growth of B. cinerea was significant and reached levels of 87.32%. Scanning electron microscopy observations revealed that after 48 h of PCN treatment, the mycelia appeared loose, locally twisted, and folded, with exudation of contents; the mycelia was withered and twisted, with edge burrs, deformations, ruptures and a sheet-like structure. Transmission electron microscopy observations revealed that after 48 h of PCN treatment, the structure of the cell nucleus was unclear and the vacuoles had ruptured; additionally, various organelles exhibited disordered structures, there were substantial non-membrane transparent inclusions, the cells were plasmolysed, the cell walls were collapsed in some cases, and the hyphal tissue was essentially necrotic. A PCN

  2. Control Effect and Possible Mechanism of the Natural Compound Phenazine-1-Carboxamide against Botrytis cinerea.

    Directory of Open Access Journals (Sweden)

    Ya Zhang

    Full Text Available To develop new agents against strawberry grey mould and to aid in the development of biological pesticides, we investigated the inhibitory effect of a natural compound, phenazine-1-carboxamide (PCN, against Botrytis cinerea using a growth rate assay. Additionally, indoor toxicity and the in vitro control effect of PCN were further studied to determine its potential mechanisms of action on B. cinerea. PCN was inhibitory against B. cinerea with a 50% effective concentration (EC50 of 108.12 μg/mL; the toxicity of PCN was equivalent to that of carbendazim (CBM. The best in vitro control effect of PCN against grey mould in strawberry (fruit reached 75.32%, which was slightly higher than that of CBM. The field control effect of PCN against grey mould reached a maximum of 72.31% at a PCN concentration of 700 μg/mL, which was 1.02 times higher than that of CBM. Fungistatic activity was observed at low concentrations of PCN, while high concentrations of PCN resulted in fungicidal activity against B. cinerea. This natural compound strongly inhibited both spore and sclerotium germination of B. cinerea, with the best relative inhibition rates of 77.03% and 82.11%, respectively. The inhibitory effect of PCN on mycelial growth of B. cinerea was significant and reached levels of 87.32%. Scanning electron microscopy observations revealed that after 48 h of PCN treatment, the mycelia appeared loose, locally twisted, and folded, with exudation of contents; the mycelia was withered and twisted, with edge burrs, deformations, ruptures and a sheet-like structure. Transmission electron microscopy observations revealed that after 48 h of PCN treatment, the structure of the cell nucleus was unclear and the vacuoles had ruptured; additionally, various organelles exhibited disordered structures, there were substantial non-membrane transparent inclusions, the cells were plasmolysed, the cell walls were collapsed in some cases, and the hyphal tissue was essentially

  3. Plant signalling components EDS1 and SGT1 enhance disease caused by the necrotrophic pathogen Botrytis cinerea.

    Science.gov (United States)

    El Oirdi, Mohamed; Bouarab, Kamal

    2007-01-01

    * Botrytis cinerea is a necrotrophic fungus that causes grey mould on a wide range of food plants, especially grapevine, tomato, soft fruits and vegetables. This disease brings about important economic losses in both pre- and postharvest crops. Successful protection of host plants against this pathogen is severely hampered by a lack of resistance genes in the hosts and the considerable phenotypic diversity of the fungus. * The aim of this study was to test whether B. cinerea manipulates the immunity-signalling pathways in plants to restore its disease. * We showed that B. cinerea caused disease in Nicotiana benthamiana through the activation of two plant signalling genes, EDS1 and SGT1, which have been shown to be essential for resistance against biotrophic pathogens; and more interestingly, virus-induced gene silencing of these two plant signalling components enhanced N. benthamiana resistance to B. cinerea. Finally, plants expressing the baculovirus antiapoptotic protein p35 were more resistant to this necrotrophic pathogen than wild-type plants. * This work highlights a new strategy used by B. cinerea to establish disease. This information is important for the design of strategies to improve plant pathogen resistance.

  4. Field and laboratory studies of the susceptibility of the green treefrog (Hyla cinerea to Batrachochytrium dendrobatidis infection.

    Directory of Open Access Journals (Sweden)

    Laura A Brannelly

    Full Text Available Amphibians worldwide are experiencing devastating declines, some of which are due to the amphibian chytrid fungus (Batrachochytrium dendrobatidis, Bd. Populations in the southeastern United States, however, have not been noticeably affected by the pathogen. The green treefrog (Hyla cinerea is abundant and widespread in the southeastern United States, but has not been documented to harbor Bd infection. This study examined the susceptibility of H. cinerea to two strains of Bd in the lab and the prevalence of infection in wild populations of this species in southeastern Louisiana. Although we were able to infect H. cinerea with Bd in the lab, we did not observe any clinical signs of chytridiomycosis. Furthermore, infection by Bd does not appear to negatively affect body condition or growth rate of post-metamorphic individuals. We found no evidence of infection in surveys of wild H. cinerea. Our results suggest that H. cinerea is not susceptible to chytridiomycosis post-metamorphosis and probably is not an important carrier of the fungal pathogen Bd in the southeastern United States, although susceptibility at the larval stage remains unknown.

  5. Botrytis californica, a new cryptic species in the B. cinerea species complex causing gray mold in blueberries and table grapes.

    Science.gov (United States)

    Saito, S; Margosan, D; Michailides, T J; Xiao, C L

    2016-01-01

    The Botrytis cinerea species complex comprises two cryptic species, originally referred to Group I and Group II based on Bc-hch gene RFLP haplotyping. Group I was described as a new cryptic species B. pseudocinerea During a survey of Botrytis spp. causing gray mold in blueberries and table grapes in the Central Valley of California, six isolates, three from blueberries and three from table grapes, were placed in Group I but had a distinct morphological character with conidiophores significantly longer than those of B. cinerea and B. pseudocinerea We compared these with B. cinerea and B. pseudocinerea by examining morphological and physiological characters, sensitivity to fenhexamid and phylogenetic analysis inferred from sequences of three nuclear genes. Phylogenetic analysis with the three partial gene sequences encoding glyceraldehyde-3-phosate dehydrogenase (G3PDH), heat-shock protein 60 (HSP60) and DNA-dependent RNA polymerase subunit II (RPB2) supported the proposal of a new Botrytis species, B. californica, which is closely related genetically to B. cinerea, B. pseudocinerea and B. sinoviticola, all known as causal agents of gray mold of grapes. Botrytis californica caused decay on blueberry and table grape fruit inoculated with the fungus. This study suggests that B. californica is a cryptic species sympatric with B. cinerea on blueberries and table grapes in California.

  6. Impedance of the Grape Berry Cuticle as a Novel Phenotypic Trait to Estimate Resistance to Botrytis Cinerea

    Directory of Open Access Journals (Sweden)

    Katja Herzog

    2015-05-01

    Full Text Available Warm and moist weather conditions during berry ripening provoke Botrytis cinerea (B. cinerea causing notable bunch rot on susceptible grapevines with the effect of reduced yield and wine quality. Resistance donors of genetic loci to increase B. cinerea resistance are widely unknown. Promising traits of resistance are represented by physical features like the thickness and permeability of the grape berry cuticle. Sensor-based phenotyping methods or genetic markers are rare for such traits. In the present study, the simple-to-handle I-sensor was developed. The sensor enables the fast and reliable measurement of electrical impedance of the grape berry cuticles and its epicuticular waxes (CW. Statistical experiments revealed highly significant correlations between relative impedance of CW and the resistance of grapevines to B. cinerea. Thus, the relative impedance Zrel of CW was identified as the most important phenotypic factor with regard to the prediction of grapevine resistance to B. cinerea. An ordinal logistic regression analysis revealed a R2McFadden of 0.37 and confirmed the application of Zrel of CW for the prediction of bunch infection and in this way as novel phenotyping trait. Applying the I-sensor, a preliminary QTL region was identified indicating that the novel phenotypic trait is as well a valuable tool for genetic analyses.

  7. Characterization of Neisseria cinerea, a nonpathogenic species isolated on Martin-Lewis medium selective for pathogenic Neisseria spp.

    Science.gov (United States)

    Knapp, J S; Totten, P A; Mulks, M H; Minshew, B H

    1984-01-01

    An asaccharolytic, gram-negative, oxidase-positive diplococcus was isolated on Martin-Lewis medium from the cervix of a patient attending an arthritis clinic at Seattle Public Health Hospital, Seattle, Wash. This strain, NRL 32165, did not produce detectable acid from glucose, maltose, sucrose, fructose, mannitol, or lactose in either cystine Trypticase agar (BBL Microbiology Systems, Cockeysville, Md.) or modified oxidation-fermentation medium and was identified presumptively as a glucose-negative Neisseria gonorrhoeae strain, but was identified later as Neisseria cinerea on the basis of its biochemical reactions. Nitrate was not reduced, nitrite (0.001%, wt/vol) was reduced, and polysaccharide was not produced from sucrose. Proline, arginine, and cystine-cysteine were required for growth on defined medium. Strain NRL 32165 did not react with antigonococcal protein I monoclonal antibodies and did not produce immunoglobulin A protease. In DNA:DNA homology studies with N. gonorrhoeae NRL 8038 (F62) and N. cinerea type strain NRL 30003, strain NRL 32165 showed 95% homology relative to N. cinerea and 44% homology relative to N. gonorrhoeae. Thus, the identity of strain NRL 32165 was confirmed as N. cinerea (von Lingelsheim 1906) Murray 1939. Of all Neisseria spp., N. cinerea is most likely to be misidentified as a glucose-negative N. gonorrhoeae strain.

  8. The ABC transporter BcatrB from Botrytis cinerea exports camalexin and is a virulence factor on Arabidopsis thaliana.

    Science.gov (United States)

    Stefanato, Francesca L; Abou-Mansour, Eliane; Buchala, Antony; Kretschmer, Matthias; Mosbach, Andreas; Hahn, Matthias; Bochet, Christian G; Métraux, Jean-Pierre; Schoonbeek, Henk-jan

    2009-05-01

    Arabidopsis thaliana is known to produce the phytoalexin camalexin in response to abiotic and biotic stress. Here we studied the mechanisms of tolerance to camalexin in the fungus Botrytis cinerea, a necrotrophic pathogen of A. thaliana. Exposure of B. cinerea to camalexin induces expression of BcatrB, an ABC transporter that functions in the efflux of fungitoxic compounds. B. cinerea inoculated on wild-type A. thaliana plants yields smaller lesions than on camalexin-deficient A. thaliana mutants. A B. cinerea strain lacking functional BcatrB is more sensitive to camalexin in vitro and less virulent on wild-type plants, but is still fully virulent on camalexin-deficient mutants. Pre-treatment of A. thaliana with UV-C leads to increased camalexin accumulation and substantial resistance to B. cinerea. UV-C-induced resistance was not seen in the camalexin-deficient mutants cyp79B2/B3, cyp71A13, pad3 or pad2, and was strongly reduced in ups1. Here we demonstrate that an ABC transporter is a virulence factor that increases tolerance of the pathogen towards a phytoalexin, and the complete restoration of virulence on host plants lacking this phytoalexin.

  9. Synthesis, Fungicidal Activity and Mode of Action of 4-Phenyl-6-trifluoromethyl-2-aminopyrimidines against Botrytis cinerea

    Directory of Open Access Journals (Sweden)

    Chunhui Liu

    2016-06-01

    Full Text Available Anilinopyrimidines are the main chemical agents for management of Botrytis cinerea. However, the drug resistance in fungi against this kind of compounds is very serious. To explore new potential fungicides against B. cinerea, a series of 4-phenyl-6-trifluoromethyl-2-amino-pyrimidine compounds (compounds III-1 to III-22 were synthesized, and their structures were confirmed by 1H-NMR, IR and MS. Most of these compounds possessed excellent fungicidal activity. The compounds III-3 and III-13 showed higher fungicidal activity than the positive control pyrimethanil on fructose gelatin agar (FGA, and compound III-3 on potato dextrose agar (PDA indicated high activity compared to the positive control cyprodinil. In vivo greenhouse results indicated that the activity of compounds III-3, III-8, and III-11 was significantly higher than that of the fungicide pyrimethanil. Scanning electron micrography (SEM and transmission electron micrography (TEM were applied to illustrate the mechanism of title compounds against B. cinerea. The title compounds, especially those containing a fluorine atom at the ortho-position on the benzene ring, could maintain the antifungal activity against B. cinerea, but their mechanism of action is different from that of cyprodinil. The present study lays a good foundation for us to find more efficient reagents against B. cinerea.

  10. Interaction with Penicillium expansum enhances Botrytis cinerea growth in grape juice medium and prevents patulin accumulation in vitro.

    Science.gov (United States)

    Morales, H; Paterson, R R M; Venâncio, A; Lima, N

    2013-05-01

    Interactions between fungi occur when they grow on the same host plant. This is the case of Botrytis cinerea and Penicillium expansum on grape. P. expansum is also responsible for production of the mycotoxin patulin. In this study, the influence of the interaction between both fungi on fungal growth parameters was studied as well as the effect on the accumulation of patulin by P. expansum. For that purpose, spores of B. cinerea and P. expansum were inoculated together (mixed inoculum), and the parameters growth rate, time for growth and patulin accumulation were assessed. The presence of P. expansum conidia shortened the time for growth of mixed inoculum colonies which, at the end of incubation, were B. cinerea-like. Although some P. expansum growth was observed in mixed inoculum colonies, very low levels of patulin were observed. In assays carried out in patulin-spiked medium, B. cinerea was capable to metabolize the mycotoxin. The capabilities of B. cinerea to shorten time for growth and prevent patulin accumulation are competing abilities that facilitate grape colonization.

  11. Effects of the origins of Botrytis cinerea on earthy aromas from grape broth media further inoculated with Penicillium expansum.

    Science.gov (United States)

    Morales-Valle, H; Silva, L C; Paterson, R R M; Venâncio, A; Lima, N

    2011-08-01

    Earthy "off" aromas from wine and grape juice are highly detrimental to the production of quality grape products. These volatile compounds are produced on grapes by Botrytis cinerea, Penicillium expansum and/or a combination of P. expansum and B. cinerea strains. B. cinerea strains were isolated from different (a) vineyards in Spain and Portugal, (b) grape varieties (c) bunches (i.e., sound and botrytized) and (d) positions in the botrytized bunch (i.e., interior or exterior). A novel Headspace-Phase Microextraction (SPME) followed by Gas Chromatrography/Mass Spectrometry (GC-MS) dedicated to analyze geosmin, methylisoborneol (MIB), 1-octen-3-ol, fenchone and fenchol in grape broth medium was used. Approximately 50% of the B. cinerea strains induced detectable geosmin. One strain accumulated significant amounts of anisoles, demonstrating that this contamination might already occur in the vineyard. Strains from the interior of Cainho grape bunches induced more geosmin and hence it may be possible to reduce this volatile in wine by avoiding using these grapes in case of B. cinerea attack.

  12. Physcomitrella patens activates reinforcement of the cell wall, programmed cell death and accumulation of evolutionary conserved defense signals...upon Botrytis cinerea infection

    Science.gov (United States)

    The moss Physcomitrella patens is an evolutionarily basal model system suitable to analyze plant defense responses activated after pathogen assault. Upon infection with the necrotroph Botrytis cinerea (B. cinerea), several defense mechanisms are induced in P. patens, including the fortification of t...

  13. Cloning of a thermostable xylanase from Actinomadura sp. S14 and its expression in Escherichia coli and Pichia pastoris.

    Science.gov (United States)

    Sriyapai, Thayat; Somyoonsap, Peechapack; Matsui, Kenji; Kawai, Fusako; Chansiri, Kosum

    2011-05-01

    A thermophilic xylan-degrading Actinomadura sp. S14 was isolated from compost in Thailand. Hemicellulase activities such as endo-1,4-β-xylanase, β-xylosidase and α-arabinofuranosidase were induced with xylan-containing agriculture wastes and oat spelt xylan. The gene encoding xylanase consisting of 687bp was cloned from Actinomadura sp. S14. The deduced amino acid sequence contained a signal peptide of 41 amino acids and a probable mature xylanase of 188 amino acids. An open reading frame (xynS14) corresponding to a mature xylanase was expressed in Escherichia coli and Pichia pastoris. The specific activity of purified XynS14 (P. pastoris) was 2.4-fold higher than XynS14 (E. coli). Both XynS14s showed the same basic properties such as optimal pH and temperature (pH 6.0 and 80°C) and stability in a broad pH range (pH 5.0-11.0) and at high temperatures up to 80°C. Both XynS14s showed approximately the same substrate specificity and K(m) values toward various xylans, but XynS14 (P. pastoris) showed higher V(max) and K(cat) than XynS14 (E. coli). Higher specific activities of XynS14 (P. pastoris) may be due to protein-folding in the host. Purified XynS14 showed more endo-1,4-β-xylanase activity on xylan and xylooligosaccharides than on xylotriose.

  14. Xylan-binding xylanase Xyl30 from Streptomyces avermitilis: cloning, characterization, and overproduction in solid-state fermentation.

    Science.gov (United States)

    Hernández, Alberto; López, José C; Santamaría, Ramón; Díaz, Margarita; Fernández-Abalos, José M; Copa-Patiño, José L; Soliveri, Juan

    2008-06-01

    A DNA fragment from the lignocellulolytic actinomycete Streptomyces avermitilis CECT 3339 was cloned using a DNA probe from the xylanase gene xysA of Streptomyces halstedii. The nucleotide sequence analysis revealed two potential ORFs, xyl30 and hd30, encoding a deduced multimodular F/10 xylanase with a binding domain and a secreted glycoxyl hydrolase, respectively. In Streptomyces lividans carrying the subcloned DNA fragment, two xylanase activity bands with estimated molecular masses of 42.8 and 35 kDa (named Xyl30 forms "h" and "l", respectively), were detected by zymograms and SDS-PAGE. The two xylanases had identical N-terminal sequences, suggesting that Xyl30 "l" derived from Xyl30 "h" by C-terminal processing in the culture supernatant. No transcripts of hd30 were detected by RT-PCR. Characterization of the partially purified Xyl30 "h" confirmed the presence of a modular endoxylanase containing a xylan-binding domain, which after processing in the culture supernatant loses the aforementioned domain and thus its capacity to bind xylan (Xyl30 "l"). Xyl30 "h" achieved maximal activity at pH 7.5 and 60 degrees C, retaining more than 50% of its activity from pH 3 to 9 and more than 40% after a 1-h incubation at 70 masculineC. Moreover, in the recombinant host strain up to 400 U xylanase/g medium (dry weight) was produced in solid-state fermentation (SSF) using cereal bran as substrate. The high production yields of this enzyme and its biochemical features make it a good candidate for use in industrial applications.

  15. Energy and nutrient utilization of broiler chickens fed corn-soybean meal and corn-based diets supplemented with xylanase.

    Science.gov (United States)

    Stefanello, C; Vieira, S L; Carvalho, P S; Sorbara, J O B; Cowieson, A J

    2016-08-01

    A study was conducted to evaluate the effects of increased levels of a β-xylanase on energy and nutrient utilization of broiler chickens fed corn-soy diets. A total of 480 slow feathering Cobb × Cobb 500 male broilers were randomly distributed to 10 treatments having 8 replicates of 6 birds each. Birds were fed a common starter diet to d 14 post hatch (3,050 kcal/kg AMEn, 21.7% CP, 1.05% Ca, and 0.53% nPP). The experimental diets were provided afterwards until 25 d. Two experimental diets, a conventional corn/soy-based basal diet (CS) and the basal diet in which 40% of the diet was displaced by corn (CN), were fed as-is or supplemented with 50, 100, 150, or 200 fungal β-xylanase units (FXU)/kg. Dietary treatments were distributed factorially as a 2 × 5 arrangement. Samples of feed, excreta, and ileal digesta were analyzed for determination of ileal digestible energy (IDE), metabolizable energy, and total tract retention of protein and lipid. No interactions between diet and xylanase were observed. The CS diets had higher (P energy utilization and nutrient digestibility when compared to the CN diets. AMEn and IDE were improved (P energy utilization and digestibility of crude protein and dry matter increased with xylanase supplementation in corn/soy-based diets. When xylanase was tested in the CS diet, 92 and 124 FXU/kg maximized the energy release effect; however, the maximum energy response in the CN diet or corn was not achieved until 200 FXU/kg.

  16. Utilization of Agro-industrial Wastes for the Simultaneous Production of Amylase and Xylanase by Thermophilic Actinomycetes

    Directory of Open Access Journals (Sweden)

    Renu Singh

    2012-12-01

    Full Text Available Agro-industrial wastes such as sugarcane bagasse, wheat bran, rice bran, corn cob and wheat straw are cheapest and abundantly available natural carbon sources. The present study was aimed to production of amylase and xylanase simultaneously using agro-industrial waste as the sole carbon source. Seven thermophilic strains of actinomycete were isolated from the mushroom compost. Among of these, strain designated MSC702 having high potential to utilize agro-industrial wastes for the production of amylase and xylanase. Strain MSC702 was identified as novel species of Streptomyces through morphological characterization and 16S rRNA gene sequence. Enzyme production was determined using 1% (w/v of various agro-industrial waste in production medium containing (g/100mL: K2HPO4(0.1, (NH42SO4(0.1, NaCl (0.1, MgSO4(0.1 at pH 7.0 after incubation of 48 h at 50°C. The amylase activity (373.89 IU/mL and xylanase activity (30.15 IU/mL was maximum in rice bran. The decreasing order of amylase and xylanase activity in different type of agro-industrial wastes were found rice bran (RB > corn cob (CC > wheat bran (WB > wheat straw (WS > sugarcane bagasse (SB and rice bran (RB > wheat bran (WB > wheat straw (WS > sugarcane bagasse (SB > corn cob (CC, respectively. Mixed effect of different agro-industrial wastes was examined in different ratios. Enzyme yield of amylase and xylanase was ~1.3 and ~2.0 fold higher with RB: WB in 1:2 ratio.

  17. Microfluidic immunosensor with micromagnetic beads coupled to carbon-based screen-printed electrodes (SPCEs) for determination of Botrytis cinerea in tissue of fruits.

    Science.gov (United States)

    Fernández-Baldo, Martín A; Messina, Germán A; Sanz, Maria I; Raba, Julio

    2010-11-10

    A wide range of plant species, including economically important crops such as vegetables, ornamentals, bulbs, and fundamentally fruits, can be affected by gray mold caused by the fungal pathogen Botrytis cinerea . This paper describes the development of a microfluidic immunosensor with micromagnetic beads (MMBs) coupled to carbon-based screen-printed electrodes (SPCEs) for the rapid and sensitive quantification of B. cinerea in apple (Red Delicious), table grape (pink Moscatel), and pear (William's) tissues. The detection of B. cinerea was carried out using a competitive immunoassay method based on the use of purified B. cinerea antigens immobilized on 3-aminopropyl-modified MMBs. The total assay time was 40 min, and the calculated detection limit was 0.008 μg mL(-1). Moreover, the intra- and interassay coefficients of variation were below 7%. The developed method allowed detects B. cinerea even in asymptomatic fruits and promises to be particularly useful for application in the agricultural industry.

  18. Regulatory puzzle of xyn1 gene (xylanase1) expression in Trichoderma reesei

    Institute of Scientific and Technical Information of China (English)

    Robert L Mach; Elisabeth Würleitner; Astrid R Stricker; Roman Rauscher; Christian Wacenovsky

    2004-01-01

    @@ Xylanase 1 (Xyn1) is one of the two major representatives of the xylanase system of T. reesei; the mechanisms governing its expression were analysed throughout this study. All factors and regulatory motifs responsible for transcriptional regulation and the model of their interplay in induction and repression will be presented. Using in vivo foot printing analysis of xylan-induced and glucose repressed mycelia, we detected three adjacent nucleotide sequences contacted by DNA-binding proteins. Protection within the inverted repeat of the Cre1 (SYGGRG) consensus sequence on the non coding strand under repressing conditions is in perfect agreement with the previously reported Cre1dependent glucose repression of xyn1. Constitutive protein binding could be observed to a CCAAT-box and an inverted repeat of a 5′ GGCTAA 3′ sequence. EMSA with crude extracts from induced and repressed mycelia revealed that the latter motifs are sufficient for formation of the basal transcriptional complex under all conditions. The inverted repeat of GGCTAA closely resembles the consensus sequences of the cellulase and xylanase regulators Ace1, Ace2 and, Xyr1 (encoded by xyr1, cloned and characterised in this study) EMSA with heterologously expressed components of each factor and of the T. reesei Hap2/3/5 protein complex revealed that the basal transcriptional complex is formed by Xyr1and the Hap2/3/5. Additionally to the Cre1 mediated carbon catabolite repression a yet unknown mechanism antagonizing induction of xyn1 expression could be elucidated. Latter occurs through competition of the repressor Ace1 and Xyr1 for the GGCTAA motif. In vivo proof for the relevance of identified motifs could be given through analysis of T. reesei transformants containing correspondingly mutated versions of the xyn1 promoter fused to the A. niger goxA gene. The results indicated that the basal as well as the induction level of xyn1 gene transcription is dependent on an interaction of Xyr1with the

  19. Study on Prevention Technology of Beans Botrytis cinerea%豆角灰霉病防治技术研究

    Institute of Scientific and Technical Information of China (English)

    冯健; 冯敏; 方新; 于淼

    2012-01-01

    灰霉病是造成豆角烂荚的主要病害,灰霉病多从残花开始侵染,然后逐渐侵染豆荚,造成烂荚,植株死棵。该研究结果表明:木霉孢子稀释液进行喷施也可以有效防治灰霉病的发生。%Botrytis cinerea is primary diseases which causes rotten bean-pod,Botrytis cinerea infection starts from the flowers,and then gradually infects bean-pod,causes rotten bean-pod and plant trees to die.This study′s results showed that the trichoderma spore diluted solution spraying could effectively control the occurrence of Botrytis cinerea.

  20. An overview of the safety evaluation of the Thermomyces lanuginosus xylanase enzyme (SP 628) and the Aspergillus aculeatus xylanase enzyme (SP 578).

    Science.gov (United States)

    Bergman, A; Broadmeadow, A

    1997-01-01

    Xylanases SP 628 and SP 578 were produced by submerged fermentation of Aspergillus oryzae, containing a gene code originating from Thermomyces lanuginosus and Aspergillus aculeatus, respectively. Both enzymes were subject to the same series of toxicological tests to document their safety in use. The enzymes are to be applied as processing aids in the baking industry and in wheat starch separation. Neither enzyme was found to be mutagenic in the Salmonella typhimurium reverse mutation assay, nor did they cause chromosomal aberrations in cultured human peripheral lymphocytes. No evidence of inhalation toxicity or skin and eye irritation was found. The enzymes are not regarded as skin-sensitizers, although the Buehler test with guinea-pigs revealed a minor potential. Oral administration up to 10.0 ml/kg bw/day (equivalent to a Total Organic Solids amount of 13.3% for SP 628 and of 11.3% for SP 578) in 13-week rat studies did not show any adverse effect.

  1. Transfer of a gonococcal beta-lactamase plasmid to conjugation-deficient Neisseria cinerea strains by transformation.

    Science.gov (United States)

    Genco, C A; Clark, V L

    1988-12-01

    We have previously shown that some strains of Neisseria cinerea can serve as recipients in conjugation (Con+) with Neisseria gonorrhoeae while others cannot (Con-). To determine if a replication defect contributes to the inability of certain strains of N. cinerea to serve as recipients in conjugation, we attempted to introduce a naturally occurring gonococcal beta-lactamase plasmid into N. cinerea by transformation. Various methods were employed, and all proved unsuccessful. Since specific sequences are required for DNA uptake in transformation of N. gonorrhoeae, we constructed a number of hybrid plasmids containing N. cinerea chromosomal DNA inserted into the N. gonorrhoeae/Escherichia coli beta-lactamase shuttle vector, pLES2. When nine randomly selected plasmids with inserts were used to transform an N. cinerea strain which did not accept the gonococcal beta-lactamase plasmid by conjugation, transformants were observed with four of the hybrid plasmids. The presence of one of the hybrid plasmids, pCAG9, in transformants was confirmed by agarose gel electrophoresis, Southern hybridization, and beta-lactamase production. When an N. gonorrhoeae donor strain containing pCAG9 was used in conjugation with several N. cinerea strains, only those strains that were previously shown to act as recipients could accept and maintain pCAG9. The ability of pCAG9 and the other three hybrid plasmids to transform Con- strains demonstrates that the beta-lactamase plasmid can replicate in Con- strains, and, therefore, the Con- phenotype is due to a block in some other stage of the conjugation process.

  2. Identification of miRNAs Responsive to Botrytis cinerea in Herbaceous Peony (Paeonia lactiflora Pall. by High-Throughput Sequencing

    Directory of Open Access Journals (Sweden)

    Daqiu Zhao

    2015-09-01

    Full Text Available Herbaceous peony (Paeonia lactiflora Pall., one of the world’s most important ornamental plants, is highly susceptible to Botrytis cinerea, and improving resistance to this pathogenic fungus is a problem yet to be solved. MicroRNAs (miRNAs play an essential role in resistance to B. cinerea, but until now, no studies have been reported concerning miRNAs induction in P. lactiflora. Here, we constructed and sequenced two small RNA (sRNA libraries from two B. cinerea-infected P. lactiflora cultivars (“Zifengyu” and “Dafugui” with significantly different levels of resistance to B. cinerea, using the Illumina HiSeq 2000 platform. From the raw reads generated, 4,592,881 and 5,809,796 sRNAs were obtained, and 280 and 306 miRNAs were identified from “Zifengyu” and “Dafugui”, respectively. A total of 237 conserved and 7 novel sequences of miRNAs were differentially expressed between the two cultivars, and we predicted and annotated their potential target genes. Subsequently, 7 differentially expressed candidate miRNAs were screened according to their target genes annotated in KEGG pathways, and the expression patterns of miRNAs and corresponding target genes were elucidated. We found that miR5254, miR165a-3p, miR3897-3p and miR6450a might be involved in the P. lactiflora response to B. cinerea infection. These results provide insight into the molecular mechanisms responsible for resistance to B. cinerea in P. lactiflora.

  3. An Interspecies Comparative Analysis of the Predicted Secretomes of the Necrotrophic Plant Pathogens Sclerotinia sclerotiorum and Botrytis cinerea.

    Directory of Open Access Journals (Sweden)

    Steph Heard

    Full Text Available Phytopathogenic fungi form intimate associations with host plant species and cause disease. To be successful, fungal pathogens communicate with a susceptible host through the secretion of proteinaceous effectors, hydrolytic enzymes and metabolites. Sclerotinia sclerotiorum and Botrytis cinerea are economically important necrotrophic fungal pathogens that cause disease on numerous crop species. Here, a powerful bioinformatics pipeline was used to predict the refined S. sclerotiorum and B. cinerea secretomes, identifying 432 and 499 proteins respectively. Analyses focusing on S. sclerotiorum revealed that 16% of the secretome encoding genes resided in small, sequence heterogeneous, gene clusters that were distributed over 13 of the 16 predicted chromosomes. Functional analyses highlighted the importance of plant cell hydrolysis, oxidation-reduction processes and the redox state to the S. sclerotiorum and B. cinerea secretomes and potentially host infection. Only 8% of the predicted proteins were distinct between the two secretomes. In contrast to S. sclerotiorum, the B. cinerea secretome lacked CFEM- or LysM-containing proteins. The 115 fungal and oomycete genome comparison identified 30 proteins specific to S. sclerotiorum and B. cinerea, plus 11 proteins specific to S. sclerotiorum and 32 proteins specific to B. cinerea. Expressed sequence tag (EST and proteomic analyses showed that 246 S. sclerotiorum secretome encoding genes had EST support, including 101 which were only expressed in vitro and 49 which were only expressed in planta, whilst 42 predicted proteins were experimentally proven to be secreted. These detailed in silico analyses of two important necrotrophic pathogens will permit informed choices to be made when candidate effector proteins are selected for function analyses in planta.

  4. Effect of xylanase and cellulase supplementation on growth performance, volatile fatty acids and caecal bacteria of broiler chickens fed with palm kernel meal-based diet

    OpenAIRE

    2014-01-01

    In this study, the effect of xylanase and cellulase supplementation in palm kernel meal (PKM) based diet on growth performance, volatile fatty acids (VFAs) and the caecal bacterial populations of broiler chickens were investigated. Seventy five day old male Cobb broiler chicks were randomly allocated to three dietary treatment groups receiving T1 (20% PKM-based diet without enzyme), T2 (20% PKM-based diet with xylanase) and T3 (20% PKM-based diet with cellulase). Each enzyme was supplemented ...

  5. CARACTERIZACION DE CEPAS DE CLONOSTACHYS ROSEA PARA EL CONTROL DE BOTRYTIS CINEREA EN VIVEROS DE EUCALYPTUS GLOBULUS

    OpenAIRE

    ZALDUA FLORES, SALOME ANDREA

    2012-01-01

    Eucalyptus globulus Labill es la segunda especie forestal más importante de Chile y una de las más susceptibles a Botrytis cinerea Pers. ex Fr. Este fitopatógeno es el responsable de la enfermedad conocida como “moho gris” en más de 200 hospederos en el mundo, siendo un serio problema económico en cultivos de pre- y post-cosecha. En Chile, los problemas ocasionados por B. cinerea han sido observados en la mayoría de los viveros, con diferentes niveles de incidencia y severidad,...

  6. Production of xylanase and protease by Penicillium janthinellum CRC 87M-115 from different agricultural wastes.

    Science.gov (United States)

    Oliveira, Luciana A; Porto, Ana L F; Tambourgi, Elias B

    2006-04-01

    Five agricultural wastes were evaluated in submerged fermentation for xylanolytic enzymes production by Penicillium janthinellum. The wastes were hydrolyzed in acid medium and the liquid fraction was used for cultivation. Corn cob (55.3 U/mL) and oat husk (54.8 U/mL) were the best inducers of xylanase. Sugar cane bagasse (23.0 U/mL) and corn husk (23.8 U/mL) were moderately good, while cassava peel was negligible. Protease production was very low in all agro-industrial residues. The maximum biomass yields were 1.30 and 1.17 g/L for cassava peel and corn husk after 180 h, respectively. Xylanolytic activity showed a cell growth associated profile.

  7. Alkalistable xylanase production by alkalitolerant Paenibacillus montaniterrae RMV1 isolated from red mud.

    Science.gov (United States)

    Arora, Amita; Krishna, Pankaj; Malik, Vinita; Reddy, M Sudhakara

    2014-10-01

    The alkalitolerant and xylanolytic bacterial strain (RMV1) isolated from red mud pond was identified as Paenibacillus montaniterrae based on both biochemical and 16S rDNA sequence analysis. The RMV1 bacterial isolate produced alkalistable and thermostable endoxylanase active over a broad range of pH (4.0-11.0) and temperature (20-100 °C), with optima at 50 °C and pH 9.0 with a T1/2 of 6.7 hours at 50 °C. This is the first report on the isolation of P. montaniterrae from bauxite residue with quite a high xylanase producing ability.

  8. Xylanase Production from Trichoderma harzianum 1073 D3 with Alternative Carbon and Nitrogen Sources

    Directory of Open Access Journals (Sweden)

    Isil Seyis

    2005-01-01

    Full Text Available The effect of some natural wastes (orange pomace, orange peel, lemon pomace, lemon peel, apple pomace, pear peel, banana peel, melon peel and hazelnut shell on the production of xylanase from Trichoderma harzianum 1073 D3 has been studied and maximum activity has been observed on melon peel (26.5 U/mg of protein followed by apple pomace and hazelnut shell. Also, molasses could be used as an additional carbon source as it decreased the production time approximately by 50 %. Finally, potential alternatives of organic nitrogen source (cotton leaf and soybean residue wastes were analyzed and it was concluded that peptone could be replaced with these residues especially when economics of the process is the major objective.

  9. Production of xylooligosaccharides from forest waste by membrane separation and Paenibacillus xylanase hydrolysis

    Directory of Open Access Journals (Sweden)

    Chun-Han Ko

    2013-02-01

    Full Text Available Xylooligosaccharides (XO, derived from the alkaline (NaOH extractant of Mikania micrantha, were produced using multiple staged membrane separation and enzymatic xylanolysis. Staged nanofiltration (NMX, ultrafiltration (EUMX, and centrifugation (EMX processes for the ethanol precipitates were conducted. NMX recovered 97.26% of total xylose and removed 73.18% of sodium ions. Concentrations of total xylose were raised from 10.98 to 51.85 mg/mL by the NMX process. Recovered xylan-containing solids were hydrolyzed by the recombinant Paenibacillus xylanase. 68% XO conversions from total xylose of NMX was achieved in 24 hours. Xylopentaose (DP 5 was the major product from NMX and EMX hydrolysis. Xylohexaose (DP 6 was the major product from EUMX hydrolysis. Results of the present study suggest the applicability for XO production by nanofiltration, as NMX gave higher XO yields compared to those from a conventional ethanol-related lignocellulosic waste conversion process.

  10. The Nitrogen Availability Interferes with Mycorrhiza-Induced Resistance against Botrytis cinerea in Tomato

    Science.gov (United States)

    Sanchez-Bel, Paloma; Troncho, Pilar; Gamir, Jordi; Pozo, Maria J.; Camañes, Gemma; Cerezo, Miguel; Flors, Víctor

    2016-01-01

    Mycorrhizal plants are generally quite efficient in coping with environmental challenges. It has been shown that the symbiosis with arbuscular mycorrhizal fungi (AMF) can confer resistance against root and foliar pathogens, although the molecular mechanisms underlying such mycorrhiza-induced resistance (MIR) are poorly understood. Tomato plants colonized with the AMF Rhizophagus irregularis display enhanced resistance against the necrotrophic foliar pathogen Botrytis cinerea. Leaves from arbuscular mycorrhizal (AM) plants develop smaller necrotic lesions, mirrored also by a reduced levels of fungal biomass. A plethora of metabolic changes takes place in AMF colonized plants upon infection. Certain changes located in the oxylipin pathway indicate that several intermediaries are over-accumulated in the AM upon infection. AM plants react by accumulating higher levels of the vitamins folic acid and riboflavin, indolic derivatives and phenolic compounds such as ferulic acid and chlorogenic acid. Transcriptional analysis support the key role played by the LOX pathway in the shoots associated with MIR against B. cinerea. Interestingly, plants that have suffered a short period of nitrogen starvation appear to react by reprogramming their metabolic and genetic responses by prioritizing abiotic stress tolerance. Consequently, plants subjected to a transient nitrogen depletion become more susceptible to B. cinerea. Under these experimental conditions, MIR is severely affected although still functional. Many metabolic and transcriptional responses which are accumulated or activated by MIR such NRT2 transcript induction and OPDA and most Trp and indolic derivatives accumulation during MIR were repressed or reduced when tomato plants were depleted of N for 48 h prior infection. These results highlight the beneficial roles of AMF in crop protection by promoting induced resistance not only under optimal nutritional conditions but also buffering the susceptibility triggered by

  11. AaERF1 positively regulates the resistance to Botrytis cinerea in Artemisia annua.

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    Xu Lu

    Full Text Available Plants are sessile organisms, and they can not move away under abiotic or biotic stresses. Thus plants have evolved a set of genes that response to adverse environment to modulate gene expression. In this study, we characterized and functionally studied an ERF transcription factor from Artemisia annua, AaERF1, which plays an important role in biotic stress responses. The AaERF1 promoter had been cloned and GUS staining results of AaERF1 promoter-GUS transgenic A. annua showed that AaERF1 is expressed ubiquitiously in all organs. Several putative cis-acting elements such as W-box, TGA-box and Py-rich element, which are involved in defense responsiveness, are present in the promoter. The expression of AaERF1 can be induced vigorously by methyl jasmonate as well as by ethephon and wounding, implying that AaERF1 may activate some of the defense genes via the jasmonic acid and ethylene signaling pathways of A. annua. The results of electrophoretic mobility shift assay (EMSA and yeast one-hybrid experiments showed that AaERF1 was able to bind to the GCC box cis-acting element in vitro and in yeast. Ectopic expression of AaERF1 could enhance the expression levels of the defense marker genes PLANT DEFENSIN1.2 (PDF1.2 and BASIC CHITINASE (ChiB, and increase the resistance to Botrytis cinerea in the 35S::AaERF1 transgenic Arabidopsis. The down-regulated expression level of AaERF1 evidently reduced the resistance to B. cinerea in A. annua. The overall results showed that AaERF1 positively regulated the resistance to B. cinerea in A. annua.

  12. Cellulase and Xylanase Production by Penicillium echinulatum in Submerged Media Containing Cellulose Amended with Sorbitol.

    Science.gov (United States)

    Todero Ritter, Carla Eliana; Camassola, Marli; Zampieri, Denise; Silveira, Mauricio Moura; Dillon, Aldo José Pinheiro

    2013-01-01

    The present work investigated the use of sorbitol as a soluble carbon source, in association with cellulose, to produce cellulases and xylanases in submerged cultures of Penicillium echinulatum 9A02S1. Because cellulose is an insoluble carbon source, in cellulase production, there are some problems with rheology and oxygen transfer. The submerged fermentations containing media composed of 0, 0.25, 0.5, 0.75, and 1% (w/v) sorbitol and cellulose that were added at different times during the cultivation; 0.2% (w/v) soy bran; 0.1% (w/v) wheat bran; and a solution of salts. The highest filter paper activity (FPA) (1.95  ±  0.04 IU·mL(-1)) was obtained on the seventh day in the medium containing 0.5% (w/v) sorbitol and 0.5% (w/v) cellulose added 24 h after the start of cultivation. However, the CMCases showed an activity peak on the sixth day (9.99 ± 0.75 IU·mL(-1)) in the medium containing 0.75% (w/v) sorbitol and 0.75% (w/v) cellulose added after 12 h of cultivation. The xylanases showed the highest activity in the medium with 0.75% (w/v) sorbitol and 0.25% (w/v) cellulose added 36 h after the start of cultivation. This strategy enables the reduction of the cellulose concentration, which in high concentrations can cause rheological and oxygen transfer problems.

  13. Optimization of Xylanase Production through Response Surface Methodology by Fusarium sp. BVKT R2 Isolated from Forest Soil and Its Application in Saccharification

    Science.gov (United States)

    Ramanjaneyulu, Golla; Rajasekhar Reddy, Bontha

    2016-01-01

    Xylanses are hydrolytic enzymes with wide applications in several industries like biofuels, paper and pulp, deinking, food, and feed. The present study was aimed at hitting at high yield xylanase producing fungi from natural resources. Two highest xylanase producing fungal isolates—Q12 and L1 were picked from collection of 450 fungal cultures for the utilization of xylan. These fungal isolates—Q12 and L1 were identified basing on ITS gene sequencing analysis as Fusarium sp. BVKT R2 (KT119615) and Fusarium strain BRR R6 (KT119619), respectively with construction of phylogenetic trees. Fusarium sp. BVKT R2 was further optimized for maximum xylanase production and the interaction effects between variables on production of xylanase were studied through response surface methodology. The optimal conditions for maximal production of xylanase were sorbitol 1.5%, yeast extract 1.5%, pH of 5.0, Temperature of 32.5°C, and agitation of 175 rpm. Under optimal conditions, the yields of xylanase production by Fusarium sp. BVKT R2 was as high as 4560 U/ml in SmF. Incubation of different lignocellulosic biomasses with crude enzyme of Fusarium sp. BVKT R2 at 37°C for 72 h could achieve about 45% saccharification. The results suggest that Fusarium sp. BVKT R2 has potential applications in saccharification process of biomass. PMID:27713726

  14. The pathogenicity of different Botrytis cinerea Pers. isolates to apples and their sensitivity to benzimidazole fungicides

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    Hanna Bryk

    2013-12-01

    Full Text Available The pathogenicity of 80 isolates of Botrytis cinerea Pers. from different hosts to apple fruit was examined. Host specificity among isolates was not found. All of the isolates, independent of their derivation, caused apple fruit rot. Isolates from apple fruits showed moderate and strong pathogenicity to apple fruits. Only 1 of the 22 examined isolates showed weak pathogenicity. Tolerance to benomyl was compared among isolates obtained from apple fruits and from other hosts. It was found that 35% of isolates from apples showed resistance to benomyl. There was no correlation between the pathogenicity of isolates and their resistance to benomyl.

  15. Proctitis associated with Neisseria cinerea misidentified as Neisseria gonorrhoeae in a child.

    Science.gov (United States)

    Dossett, J H; Appelbaum, P C; Knapp, J S; Totten, P A

    1985-04-01

    An 8-year-old boy developed proctitis. Rectal swabs yielded a Neisseria sp. that was repeatedly identified by API (Analytab Products, Plainview, N.Y.), Minitek (BBL Microbiology Systems, Cockeysville, Md.), and Bactec (Johnston Laboratories, Towson, Md.) methods as Neisseria gonorrhoeae. Subsequent testing in a reference laboratory yielded an identification of Neisseria cinerea. It is suggested that identification of a Neisseria sp. isolated from genital or rectal sites in a child be confirmed by additional serological, growth, and antibiotic susceptibility tests and, if necessary, by a reference laboratory. The implications of such misidentifications are discussed.

  16. Production of xylan degrading endo-1, 4-β-xylanase from thermophilic Geobacillus stearothermophilus KIBGE-IB29

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    Zainab Bibi

    2014-10-01

    Full Text Available Xylan degrading bacterial strain was isolated from soil and identified as Geobacillus stearothermophilus KIBGE-IB29 on the basis of morphological, biochemical and 16S rDNA sequence analysis. Optimization of medium and culture conditions in submerged fermentation was investigated for maximum endo-1, 4-β-xylanase production. High yield of xylan degrading endo-1, 4-β-xylanase was achieved at 60 °C and pH-6.0 with 24 h of fermentation. Maximum enzyme was produced using 0.5% xylan as a carbon source, 0.5% peptone, 0.2% yeast extract and 0.1% meat extract as nitrogen sources. Di-potassium hydrogen phosphate (0.25%, calcium chloride (0.01%, potassium hydrogen phosphate (0.05% and ammonium sulfate (0.05% were also incorporated in the fermentation medium to enhance the enzyme production.

  17. Evaluation of operational parameters on the precipitation of endoglucanase and xylanase produced by solid state fermentation of Aspergillus niger

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    C. S. Farinas

    2011-03-01

    Full Text Available In order to develop cost effective processes for converting biomass into biofuels, it is essential to improve enzyme production yields, stability and specific activity. In this context, the aim of this work was to evaluate the concentration of two enzymes involved in the hydrolysis of biomass, endoglucanase and xylanase, through precipitation. Statistical experimental design was used to evaluate the influence of precipitant agent concentration (ammonium sulfate and ethanol, aging time, and temperature on enzyme activity recovery. Precipitant agent concentration and aging time showed a statistically significant effect at the 95% confidence level, on both enzyme activity recoveries. The recovery of endoglucanase with ammonium sulfate and ethanol reached values up to 65 and 61%, respectively. For xylanase, the recovery rates were lower, 27 and 25% with ammonium sulfate and ethanol, respectively. The results obtained allowed the selection of the variables relevant to improving enzyme activity recovery within operational conditions suitable for industrial applications.

  18. Apoplastic Nucleoside Accumulation in Arabidopsis Leads to Reduced Photosynthetic Performance and Increased Susceptibility Against Botrytis cinerea.

    Science.gov (United States)

    Daumann, Manuel; Fischer, Marietta; Niopek-Witz, Sandra; Girke, Christopher; Möhlmann, Torsten

    2015-01-01

    Interactions between plant and pathogen often occur in the extracellular space and especially nucleotides like ATP and NAD have been identified as key players in this scenario. Arabidopsis mutants accumulating nucleosides in the extracellular space were generated and studied with respect to susceptibility against Botrytis cinerea infection and general plant fitness determined as photosynthetic performance. The mutants used are deficient in the main nucleoside uptake system ENT3 and the extracellular nucleoside hydrolase NSH3. When grown on soil but not in hydroponic culture, these plants markedly accumulate adenosine and uridine in leaves. This nucleoside accumulation was accompanied by reduced photosystem II efficiency and altered expression of photosynthesis related genes. Moreover, a higher susceptibility toward Botrytis cinerea infection and a reduced induction of pathogen related genes PR1 and WRKY33 was observed. All these effects did not occur in hydroponically grown plants substantiating a contribution of extracellular nucleosides to these effects. Whether reduced general plant fitness, altered pathogen response capability or more direct interactions with the pathogen are responsible for these observations is discussed.

  19. Apoplastic nucleoside accumulation in Arabidopsis leads to reduced photosynthetic performance and increased susceptibility against Botrytis cinerea

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    Manuel eDaumann

    2015-12-01

    Full Text Available ABSTRACT Interactions between plant and pathogen often occur in the extracellular space and especially nucleotides like ATP and NAD have been identified as key players in this scenario. Arabidopsis mutants accumulating nucleosides in the extracellular space were generated and studied with respect to susceptibility against Botrytis cinerea infection and general plant fitness determined as photosynthetic performance. The mutants used are deficient in the main nucleoside uptake system ENT3 and the extracellular nucleoside hydrolase NSH3. When grown on soil but not in hydroponic culture, these plants markedly accumulate adenosine and uridine in leaves. This nucleoside accumulation was accpmpanied by reduced photosystem II efficiency and altered expression of photosynthesis related genes. Moreover, a higher susceptibility towards Botrytis cinerea infection and a reduced induction of pathogen related genes PR1 and WRKY33 was observed. All these effects did not occur in hydroponically grown plants substantiating a contribution of extracellular nucleosides to these effects. Whether reduced general plant fitness, altered pathogen response capability or more direct interactions with the pathogen are responsible for these observations is discussed.

  20. Discovery of two new inhibitors of Botrytis cinerea chitin synthase by a chemical library screening.

    Science.gov (United States)

    Magellan, Hervé; Boccara, Martine; Drujon, Thierry; Soulié, Marie-Christine; Guillou, Catherine; Dubois, Joëlle; Becker, Hubert F

    2013-09-01

    Chitin synthases polymerize UDP-GlcNAC to form chitin polymer, a key component of fungal cell wall biosynthesis. Furthermore, chitin synthases are desirable targets for fungicides since chitin is absent in plants and mammals. Two potent Botrytis cinerea chitin synthase inhibitors, 2,3,5-tri-O-benzyl-d-ribose (compound 1) and a 2,5-functionalized imidazole (compound 2) were identified by screening a chemical library. We adapted the wheat germ agglutinin (WGA) test for chitin synthase activity detection to allow miniaturization and robotization of the screen. Both identified compounds inhibited chitin synthases in vitro with IC50 values of 1.8 and 10μM, respectively. Compounds 1 and 2 were evaluated for their antifungal activity and were found to be active against B. cinerea BD90 strain with MIC values of 190 and 100μM, respectively. Finally, we discovered that both compounds confer resistance to plant leaves against the attack of the fungus by reducing the propagation of lesions by 37% and 23%, respectively. Based on the inhibitory properties found in different assays, compounds 1 and 2 can be considered as antifungal hit inhibitors of chitin synthase, allowing further optimization of their pharmacological profile to improve their antifungal properties.

  1. Purification and Phytotoxic Analysis of Botrytis cinerea Virulence Factors: New Avenues for Crop Protection

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    Maria R. Davis

    2012-07-01

    Full Text Available Botrytis cinerea is a necrotrophic fungus infecting over 230 plant species worldwide. This highly adaptable pathogen can afflict agricultural products from seed to storage, causing significant economic losses and instability in the food supply. Small protein virulence factors secreted by B. cinerea during infection play an important role in initiation and spread of disease. BcSnod1 was found to be abundantly expressed upon exposure to media containing strawberry extract. From sequence similarity, BcSnod2 was also identified and both were recognized as members of the Ceratoplatanin family of small phytotoxic proteins. Recombinant BcSnod1 was shown to have a phytotoxic effect and play an important role in pathogenicity while the role of BcSnod2 remains less clear. Both bacterial and yeast production systems are reported, though the bacterial protein is less toxic and mostly unfolded relative to that made in yeast. Compared to BcSnod1, recombinant bacterial BcSnod2 shows similar, but delayed phytotoxicity on tomato leaves. Further studies of these critical virulence factors and their inhibition promise to provide new avenues for crop protection.

  2. Phytotoxic activity and metabolism of Botrytis cinerea and structure-activity relationships of isocaryolane derivatives.

    Science.gov (United States)

    Ascari, Jociani; Boaventura, Maria Amélia Diamantino; Takahashi, Jacqueline Aparecida; Durán-Patrón, Rosa; Hernández-Galán, Rosario; Macías-Sánchez, Antonio J; Collado, Isidro G

    2013-06-28

    Research has been conducted on the biotransformation of (8S,9R)-isocaryolan-9-ol (4a) and (1S,2S,5R,8S)-8-methylene-1,4,4-trimethyltricyclo[6.2.1.0(2,5)]undecan-12-ol (5a) by the fungal phytopathogen Botrytis cinerea. The biotransformation of compound 4a yielded compounds 6-9, while the biotransformation of compound 5a yielded compounds 10-13. The activity of compounds 4a and 5a against B. cinerea has been evaluated. (8R,9R)-Isocaryolane-8,9-diol (6), a major metabolite of compound 4a, shows activity compared to its parent compound 4a, which is inactive. The effect of isocaryolanes 3, 4a, and 5a, together with their biotransformation products 6-8, 10, and 14-17, on the germination and radicle and shoot growth of Lactuca sativa (lettuce) has also been determined. Compounds 7-13 are described for the first time.

  3. Strand-Specific RNA-Seq Analyses of Fruiting Body Development in Coprinopsis cinerea.

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    Hajime Muraguchi

    Full Text Available The basidiomycete fungus Coprinopsis cinerea is an important model system for multicellular development. Fruiting bodies of C. cinerea are typical mushrooms, which can be produced synchronously on defined media in the laboratory. To investigate the transcriptome in detail during fruiting body development, high-throughput sequencing (RNA-seq was performed using cDNA libraries strand-specifically constructed from 13 points (stages/tissues with two biological replicates. The reads were aligned to 14,245 predicted transcripts, and counted for forward and reverse transcripts. Differentially expressed genes (DEGs between two adjacent points and between vegetative mycelium and each point were detected by Tag Count Comparison (TCC. To validate RNA-seq data, expression levels of selected genes were compared using RPKM values in RNA-seq data and qRT-PCR data, and DEGs detected in microarray data were examined in MA plots of RNA-seq data by TCC. We discuss events deduced from GO analysis of DEGs. In addition, we uncovered both transcription factor candidates and antisense transcripts that are likely to be involved in developmental regulation for fruiting.

  4. Synthesis of New Hydrated Geranylphenols and in Vitro Antifungal Activity against Botrytis cinerea

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    Mauricio Soto

    2016-06-01

    Full Text Available Geranylated hydroquinones and other geranylated compounds isolated from Aplydium species have shown interesting biological activities. This fact has prompted a number of studies where geranylated phenol derivatives have been synthesized in order to assay their bioactivities. In this work, we report the synthesis of a series of new hydrated geranylphenols using two different synthetic approaches and their inhibitory effects on the mycelial growth of Botrytis cinerea. Five new hydrated geranylphenols were obtained by direct coupling reaction between geraniol and phenol in dioxane/water and using BF3·Et2O as the catalyst or by the reaction of a geranylated phenol with BF3·Et2O. Two new geranylated quinones were also obtained. The synthesis and structural elucidation of all new compounds is presented. All hydrated geranylphenols efficiently inhibit the mycelial growth of B. cinerea. Their activity is higher than that observed for non-hydrated compounds. These results indicate that structural modification on the geranyl chain brings about an enhancement of the inhibition effect of geranylated phenol derivatives.

  5. STATISTICAL OPTIMIZATION OF MINERAL SALT AND UREA CONCENTRATION FOR CELLULASE AND XYLANASE PRODUCTION BY Penicillium echinulatum IN SUBMERGED FERMENTATION

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    L. dos Reis

    2015-03-01

    Full Text Available Abstract Penicillium echinulatum S1M29 is a mutant with cellulase and xylanase production comparable to the most studied microorganisms in the literature. However, its potential to produce these enzymes has not been fully investigated. This study aimed at optimizing salt and urea concentrations in the mineral solution, employing the response surface methodology. A 25-1 Fractional Factorial Design and a 23 Central Composite Design were applied to elucidate the effect of salts and urea in enzyme production. Lower concentrations of KH2PO4 (2.0 g.L-1, (NH42SO4 (1.4 g.L-1, MgSO4.7H2O (0.375 g.L-1 and CaCl2 (0.375 g.L-1 were most suitable for the production of all enzymes evaluated. Nevertheless, higher concentrations of urea (0.525 g.L-1 gave the best results for cellulase and xylanase production. The maximum FPase (1,5 U.m.L-1, endoglucanase (7,2 U.m.L-1, xylanase (30,5 U.m.L-1 and β-glucosidase (4,0 U.m.L-1 activities obtained with the planned medium were, respectively, 87, 16, 17 and 21% higher when compared to standard medium. The experimental design contributed to adjust the concentrations of minerals and urea of the culture media for cellulase and xylanase production by P. echinulatum, avoiding waste of components in the medium.

  6. STATISTICAL OPTIMIZATION OF MINERAL SALT AND UREA CONCENTRATION FOR CELLULASE AND XYLANASE PRODUCTION BY Penicillium echinulatum IN SUBMERGED FERMENTATION

    OpenAIRE

    L. dos Reis; Ritter,C. E. T.; R. C. Fontana; Camassola,M.; A. J. P. Dillon

    2015-01-01

    Abstract Penicillium echinulatum S1M29 is a mutant with cellulase and xylanase production comparable to the most studied microorganisms in the literature. However, its potential to produce these enzymes has not been fully investigated. This study aimed at optimizing salt and urea concentrations in the mineral solution, employing the response surface methodology. A 25-1 Fractional Factorial Design and a 23 Central Composite Design were applied to elucidate the effect of salts and urea in enzym...

  7. Properties of an alkali-thermo stable xylanase from Geobacillus thermodenitrificans A333 and applicability in xylooligosaccharides generation.

    Science.gov (United States)

    Marcolongo, Loredana; La Cara, Francesco; Morana, Alessandra; Di Salle, Anna; Del Monaco, Giovanni; Paixão, Susana M; Alves, Luis; Ionata, Elena

    2015-04-01

    An extracellular thermo-alkali-stable and cellulase-free xylanase from Geobacillus thermodenitrificans A333 was purified to homogeneity by ion exchange and size exclusion chromatography. Its molecular mass was 44 kDa as estimated in native and denaturing conditions by gel filtration and SDS-PAGE analysis, respectively. The xylanase (GtXyn) exhibited maximum activity at 70 °C and pH 7.5. It was stable over broad ranges of temperature and pH retaining 88 % of activity at 60 °C and up to 97 % in the pH range 7.5-10.0 after 24 h. Moreover, the enzyme was active up to 3.0 M sodium chloride concentration, exhibiting at that value 70 % residual activity after 1 h. The presence of other metal ions did not affect the activity with the sole exceptions of K(+) that showed a stimulating effect, and Fe(2+), Co(2+) and Hg(2+), which inhibited the enzyme. The xylanase was activated by non-ionic surfactants and was stable in organic solvents remaining fully active over 24 h of incubation in 40 % ethanol at 25 °C. Furthermore, the enzyme was resistant to most of the neutral and alkaline proteases tested. The enzyme was active only on xylan, showing no marked preference towards xylans from different origins. The hydrolysis of beechwood xylan and agriculture-based biomass materials yielded xylooligosaccharides with a polymerization degree ranging from 2 to 6 units and xylobiose and xylotriose as main products. These properties indicate G. thermodenitrificans A333 xylanase as a promising candidate for several biotechnological applications, such as xylooligosaccharides preparation.

  8. Identification of glutamic acid 78 as the active site nucleophile in Bacillus subtilis xylanase using electrospray tandem mass spectrometry.

    Science.gov (United States)

    Miao, S; Ziser, L; Aebersold, R; Withers, S G

    1994-06-14

    A new mechanism-based inactivator of beta-1,4-xylanases, 2',4'-dinitrophenyl 2-deoxy-2-fluoro-beta-xylobioside, has been synthesized and used to trap the covalent intermediate formed during catalysis by Bacillus subtilis xylanase. Electrospray mass spectrometry confirmed the 1:1 stoichiometry of the incorporation of inactivator into the enzyme. Inactivation of xylanase followed the expected pseudo-first-order kinetic behavior, and kinetic parameters were determined. The intermediate trapped was relatively stable toward hydrolytic turnover (t1/2 = 350 min). However, turnover could be facilitated by transglycosylation following the addition of the acceptor benzyl thio-beta-xylobioside, thus demonstrating the catalytic competence of the trapped intermediate. Reactivation kinetic parameters for this process of kre = 0.03 min-1 and Kre = 46 mM were determined. The nucleophilic amino acid was identified as Glu78 by a tandem mass spectrometric technique which does not require the use of radiolabels. The peptic digest of the labeled enzyme was separated by high-performance liquid chromatography and the eluent fed into a tandem mass spectrometer via an electrospray ionization device. The labeled peptide was identified as one of m/z = 826 (doubly charged) which fragmented in the collision chamber between the mass analyzers with loss of the mass of a 2-fluoroxylobiosyl unit. Confirmation of the peptide identity was obtained both by tandem mass spectrometric sequencing and by Edman degradation of the purified peptide. Glu78 is completely conserved in all members of this xylanase family and indeed is shown to be located in the active site in the recently determined X-ray crystal structure.

  9. Role of temperature and free moisture in onion flower blight. [Botrytis squamosa; Botrytis cinerea; and Botrytis allii

    Energy Technology Data Exchange (ETDEWEB)

    Ramsey, G.R.; Lorbeer, J.W.

    1986-06-01

    The cardinal temperatures at which onion umbels were blighted (after inoculation when two-thirds of the florets were open) with Botrytis squamosa, B. cinerea, and B. allii (isolated from blighted onion florets) were near 9, 21, and 27 C for B. squamosa, near 12, 21, and 30 C for B. cinerea, and near 9, 24, and 30 C for B. allii. The cardinal temperatures for mycelial growth (potato-dextrose agar) of B. squamosa, B. cinerea, and B. allii were near 5, 22, and 30 C for each fungus. The cardinal temperatures for conidial germination (on purified water agar) were near 6, 15, and 30 C for B. squamosa; 3, 18, and 33 C for B. cinerea; and 6, 24, and 33 C for B. allii. When the duration of free moisture on umbels after inoculation with the three pathogens was increased from 0 to 96 hr. the percentages of unopened florets, open florets, and immature seed capsules blighted at 21 C were increased significantly. Free moisture durations of 12-24, 6-12, and 6-12 hr were necessary for blighting of unopen florets, open florets, and immature seed capsules, respectively, by each pathogen at 21 C. A positive correlation between the amount of July rainfall and the natural incidence of onion flower blight was observed in Orange County, New York, from 1976 to 1981. 10 references, 2 figures, 1 table.

  10. Unraveling the in vitro secretome of the phytopathogen Botrytis cinerea to understand the interaction with its hosts

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    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.

  11. Emerging trends in molecular interactions between plants and the broad host range fungal pathogens Botrytis cinerea and Sclerotinia sclerotiorum

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    Malick eMbengue

    2016-03-01

    Full Text Available Fungal plant pathogens are major threats to food security worldwide. Sclerotinia sclerotiorum and Botrytis cinerea are closely related Ascomycete plant pathogens causing mold diseases on hundreds of plant species. There is no genetic source of complete plant resistance to these broad host range pathogens known to date. Instead, natural plant populations show a continuum of resistance levels controlled by multiple genes, a phenotype designated as quantitative disease resistance. Little is known about the molecular mechanisms controlling the interaction between plants and S. sclerotiorum and B. cinerea but significant advances were made on this topic in the last years. This minireview highlights a selection of nine themes that emerged in recent research reports on the molecular bases of plant-S. sclerotiorum and plant-B. cinerea interactions. On the fungal side, this includes progress on understanding the role of oxalic acid, on the study of fungal small secreted proteins. Next, we discuss the exchanges of small RNA between organisms and the control of cell death in plant and fungi during pathogenic interactions. Finally on the plant side, we highlight defense priming by mechanical signals, the characterization of plant Receptor-like proteins and the hormone abscisic acid in the response to B. cinerea and S. sclerotiorum , the role of plant general transcription machinery and plant small bioactive peptides. These represent nine trends we selected as remarkable in our understanding of fungal molecules causing disease and plant mechanisms associated with disease resistance to two devastating broad host range fungi.

  12. Physiological variability and in vitro antifungal activity against Botrytis cinerea causing botrytis gray mold of chickpea (Cicer arietinum L.)

    Energy Technology Data Exchange (ETDEWEB)

    Hosen, M. I.; Ahmed, A. U.; Islam, M. R.

    2010-07-01

    Physiological variability was studied in 10 isolates of Botrytis cinerea causing botrytis gray mold of chickpea, collected from diverse agro climatic areas in Bangladesh. The optimum temperature and pH for the best mycelial radial growth of B. cinerea were 20 degree centigrade and 4.5, respectively. The mycelial radial growth increased with the temperature up to 20 degree centigrade thereafter it decreased gradually up to 30 degree centigrade and no growth was observed at 35 degree centigrade. Chickpea dextrose agar (CDA) medium supported the highest mycelial radial growth (79.17 mm). The quickest (in 5 days) sclerotia initiation was recorded on chickpea destrose agar and lentil dextrose agar (LDA) culture media while the highest number of spores (2.5104 mL{sup -}1) were recorded on LDA medium. The antagonist Trichoderma harzianum was found to be a good bio-control agent against B. cinerea. Among the seven fungicides Bavistin 50 WP (Carbendazim), CP-Zim 50 WP (Carbendazim), Sunphanate 70 WP (Thiophanate methyl) and Rovral 50 WP (Iprodione) were the most effective to inhibit the mycelial radial growth of B. cinerea at 500 mg L{sup -}1 concentration. (Author) 13 refs.

  13. The ABC transporter BcatrB affects the sensitivity of Botrytis cinerea to the phytoalexin resveratrol and the fungicide fenpiclonil

    NARCIS (Netherlands)

    Schoonbeek, H.; Sorbo, Del G.; Waard, De M.A.

    2001-01-01

    During pathogenesis, fungal pathogens are exposed to a variety of fungitoxic compounds. This may be particularly relevant to Botrytis cinerea, a plant pathogen that has a broad host range and, consequently, is subjected to exposure to many plant defense compounds. In practice, the pathogen is contro

  14. Ameliorative potential of Vernonia cinerea on chronic constriction injury of sciatic nerve induced neuropathic pain in rats

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    VENKATA R.K. THIAGARAJAN

    2014-09-01

    Full Text Available The aim of the present study is to investigate the ameliorative potential of ethanolic extract of whole plant of Vernonia cinerea in the chronic constriction injury (CCI of sciatic nerve induced neuropathic pain in rats. Behavioral parameters such as a hot plate, acetone drop, paw pressure, Von Frey hair and tail immersion tests were performed to assess the degree of thermal, chemical and mechanical hyperalgesia and allodynia. Biochemical changes in sciatic nerve tissue were ruled out by estimating thiobarbituric acid reactive substances (TBARS, reduced glutathione (GSH and total calcium levels. Ethanolic extract of Vernonia cinerea and pregabalin were administered for 14 consecutive days starting from the day of surgery. CCI of sciatic nerve has been shown to induce significant changes in behavioral, biochemical and histopathological assessments when compared to the sham control group. Vernonia cinerea attenuated in a dose dependent manner the above pathological changes induced by CCI of the sciatic nerve, which is similar to attenuation of the pregabalin pretreated group. The ameliorating effect of ethanolic extract of Vernonia cinerea against CCI of sciatic nerve induced neuropathic pain may be due to the presence of flavonoids and this effect is attributed to anti-oxidative, neuroprotective and calcium channel modulator actions of these compounds.

  15. Fungicide-driven evolution and molecular basis of multidrug resistance in field populations of the grey mould fungus Botrytis cinerea

    NARCIS (Netherlands)

    Kretschmer, M.; Leroch, M.; Mosbach, A.; Walker, A.S.; Fillinger, S.; Mernke, D.; Schoonbeek, H.J.; Pradier, J.M.; Leroux, P.; Waard, de M.A.; Hahn, M.

    2009-01-01

    The grey mould fungus Botrytis cinerea causes losses of commercially important fruits, vegetables and ornamentals worldwide. Fungicide treatments are effective for disease control, but bear the risk of resistance development. The major resistance mechanism in fungi is target protein modification res

  16. Functional analysis of ABC transporter genes from Botrytis cinerea identifies BcatrB as a transporter of eugenol

    NARCIS (Netherlands)

    Schoonbeek, H.; Nistelrooy, van J.G.M.; Waard, de M.A.

    2003-01-01

    The role of multiple ATP-binding cassette (ABC) and major facilitator superfamily (MFS) transporter genes from the plant pathogenic fungus Botrytis cinerea in protection against natural fungitoxic compounds was studied by expression analysis and phenotyping of gene-replacement mutants. The expressio

  17. Botrytis californica, a new cryptic species in the B. cinerea species complex causing gray mold in blueberries and table grapes

    Science.gov (United States)

    Botrytis cinerea consists of two cryptic species, referred to as Group I and Group II based on Bc-hch gene RFLP haplotyping, and Group I has been described as a new cryptic species B. pseudocinerea. During a survey for Botrytis spp. causing gray mold in blueberries and table grapes in the Central Va...

  18. Effect of gamma irradiation and its convergent treatment for control of postharvest Botrytis cinerea of cut roses

    Science.gov (United States)

    Chu, Eun-Hee; Shin, Eun-Jung; Park, Hae-Jun; Jeong, Rae-Dong

    2015-10-01

    Postharvest diseases cause considerable losses to harvested crops. Among them, gray mold (Botrytis cinerea) is a major problem of exporting to cut rose flowers into Korea. Irradiation treatment is an alternative to phytosanitary purposes and a useful nonchemical approach to the control of postharvest diseases. Gamma irradiation was evaluated for its in vitro and in vivo antifungal activity against B. cinerea on cut rose varieties, 'Shooting Star' and 'Babe'. The irradiating dose required to reduce the population by 90%, D10, was 0.99 kGy. Gamma irradiation showed complete inhibition of spore germination and mycelial growth of B. cinerea, especially 4.0 kGy in vitro. Antifungal activity of gamma irradiation on rose B. cinerea is a dose-dependent manner. A significant phytotoxicity such as bent neck in cut rose quality was shown from gamma irradiation at over 0.4 kGy (pblue mold decay in both varieties. Together, these results suggest that a synergistic effect of the combined treatment with gamma irradiation and NaDCC can be efficiently used to control the postharvest diseases in cut rose flowers, and will provide a promising technology for horticulture products for exportation.

  19. Trichothecenes and aspinolides produced by Trichoderma arundinaceum regulate expression of Botrytis cinerea genes involved in virulence and growth

    Science.gov (United States)

    Trichoderma arundinaceum (Ta37) and Botrytis cinerea (B05.10) produce the sesquiterpenoid compounds harzianum A (HA) and botrydial (BOT), respectively. T. arundinaceum Ta(delta)Tri5, a mutant that does not produce HA, produces high levels of the polyketide compounds aspinolide (Asp) B and C. We anal...

  20. Botrydial and botcinins produced by Botrytis cinerea regulate expression of Trichoderma arundinaceum genes involved in trichothecene biosynthesis

    Science.gov (United States)

    Trichoderma arundinaceum (Ta37) and Botrytis cinerea produce the sesquiterpenes harzianum A (HA) and botrydial (BOT), respectively, and also the polyketides aspinolides (Asp) and botcinines (Botc), respectively. In the present work, we analyzed the role of BOT and Botcs in the T. arundinaceum-B. cin...

  1. Effect of temperature on the morphological characteristics of Botrytis cinerea and its correlated with the genetic variability

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    Jorge G Fernández

    2014-07-01

    Full Text Available Objective: To study the effect of temperature on the morphological characteristics of Botrytis cinerea (B. cinerea and its correlated with the genetic variability. B. cinerea is a plant-pathogenic fungus that produces the disease known as grey mould in a wide variety of agriculturally important hosts in many countries. Methods: Six strains from different host collected have been isolated and characterized by several methods as mycelial growth, fungicide resistance, pathogenicity and the effects of the temperature. Also was analyzed by PCR and distinguished by the presence or absence of transposable elements. Results: Results showed that clear morphological differences exist between strains at the temperature of 4, 12 and 28 °C. All strains analyzed molecularly were classified as Group II (transposa-type. Demonstrating a negative correlation between mycelial growth and other characteristics as the fungicide resistance and pathogenicity. Lastly, it is difficult to establish relationships phenotypic and genotypic between strains of B. cinerea. Conclusions: The results indicated that the mycelial growth, resistance at fungicide and pathogenicity are independent of the characteristics molecular, however, are dependent of a factor such as temperature.

  2. In vitro studies on the effect of some chemicals on the growth and sporification of Penicillium expansum and Botrytis cinerea.

    Science.gov (United States)

    Pani, G; Molinu, M G; Dore, A; Venditti, T; Petretto, A; D'Hallewin, G

    2011-01-01

    Penicillium expansum and Botrytis cinerea are among the pathogens most frequently affecting apples and grapes after harvest, respectively. We studied the behaviour of these moulds when subjected to different concentrations of methanol (MeOH) and dimethyl sulfoxide (DMSO) as a alternative method to fungicides in controlling postharvest decay of horticultural products. The experiments were performed with 5 cm Petri dishes containing PDA amended with 0, 5, 10, 20, 30, 40 or 50 microL/mL of the two tested chemicals. Freshly prepared conidia of B. cinerea and P. expansum were sown onto the media and then kept into an incubation chamber at 21 degrees C up to 3 and 6 days, respectively. Daily, the colony forming units (cfu), the colony diameter and the degree of sporification were monitored. Compared to the control, both chemicals affected the growth rate of the two pathogens. The P. expansum and B. cinerea cfu value was not significantly inhibited but the colony diameter and the sporification degree decreased when concentration was raised. B. cinerea cultured on DMSO showed a significant drop of sporification up to the tested concentration of 10 microL/mL, and a complete inhibition of cfu when the concentration was higher than 20 microL/mL.

  3. Cellulase and xylanase productions by isolated Amazon Bacillus strains using soybean industrial residue based solid-state cultivation

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    Heck Júlio X.

    2002-01-01

    Full Text Available In Brazil, a large amount of a fibrous residue is generated as result of soybean (Glycine max protein production. This material, which is rich in hemicellulose and cellulose, can be used in solid state cultivations for the production of valuable metabolites and enzymes. In this work, we studied the bioconversion of this residue by bacteria strains isolated from water and soil collected in the Amazon region. Five strains among 87 isolated bacteria selected for their ability to produce either celullases or xylanases were cultivated on the aforementioned residue. From strain BL62, identified as Bacillus subtilis, it was obtained a preparation showing the highest specific cellulase activity, 1.08 UI/mg protein within 24 hours of growth. Concerning xylanase, the isolate BL53, also identified as Bacillus subtilis, showed the highest specific activity for this enzyme, 5.19 UI/mg protein within 72 hours of cultivation. It has also been observed the production of proteases that were associated with the loss of cellulase and xylanase activities. These results indicated that the selected microorganisms, and the cultivation process, have great biotechnological potential.

  4. A xylanase from Streptomyces sp. FA1: heterologous expression, characterization, and its application in Chinese steamed bread.

    Science.gov (United States)

    Xu, Yang; Wu, Jing; Zheng, Kaixuan; Wu, Dan

    2016-05-01

    Xylanases (EC 3.2.1.8) are hydrolytic enzymes that have found widespread application in the food, feed, and paper-pulp industries. Streptomyces sp. FA1 xynA was expressed as a secreted protein in Pichia pastoris, and the xylanase was applied to the production of Chinese steamed bread for the first time. The optimal pH and the optimal temperature of XynA were 5.5 and 60 °C, respectively. Using beechwood as substrate, the K m and V max were 2.408 mg mL(-1) and 299.3 µmol min(-1) mg(-1), respectively. Under optimal conditions, a 3.6-L bioreactor produced 1374 U mL(-1) of XynA activity at a protein concentration of 6.3 g L(-1) after 132 h of fermentation. Use of recombinant XynA led to a greater increase in the specific volume of the CSB than could be achieved using commercial xylanase under optimal conditions. This study provides the basis for the application of the enzyme in the baking industry.

  5. Effect of Temperature on Xylanase II from Trichoderma reesei QM 9414: A Calorimetric, Catalytic, and Conformational Study

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    Gloria López

    2014-01-01

    Full Text Available The secondary structure of xylanase II from Trichoderma reesei is lost in an apparent irreversible cooperative process as temperature is increased with a midpoint transition of 58.8 ± 0.1°C. The shift of the spectral centre of mass above 50°C is also apparently cooperative with midpoint transition of 56.3 ± 0.2°C, but the existence of two isofluorescent points in the fluorescence emission spectra suggests a non-two-state process. Further corroboration comes from differential scanning calorimetry experiments. At protein concentrations ≤0.56 mg·mL−1 the calorimetric transition is reversible and the data were fitted to a non-two-state model and deconvoluted into six transitions, whereas at concentrations greater than 0.56 mg·mL−1 the calorimetric transition is irreversible with an exothermic contribution to the thermogram. The apparent Tm increased linearly with the scan rate according to first order inactivation kinetics. The effect of additives on the calorimetric transition of xylanase is dependent on their nature. The addition of sorbitol transforms reversible transitions into irreversible transitions while stabilizing the protein as the apparent Tm increases linearly with sorbitol concentration. d-Glucono-1,5-lactone, a noncompetitive inhibitor in xylanase kinetics, and soluble xylan change irreversible processes into reversible processes at high protein concentration.

  6. STUDIES ON XYLANASE AND LACCASE ENZYMATIC PREBLEACHING TO REDUCE CHLORINE-BASED CHEMICALS DURING CEH AND ECF BLEACHING

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    Vasanta V. Thakur,

    2012-02-01

    Full Text Available The biobleaching efficiency of xylanase and laccase enzymes was studied on kraft pulps from wood and nonwood based raw materials employed in the Indian paper industry. Treatment of these pulps with xylanase enzyme could result in improved properties, showing 2.0% ISO gain in pulp brightness and/or reducing the demand of chlorine-based bleach chemicals by up to 15% with simultaneous reduction of 20 to 25% in AOX generation in bleach effluents. Further, mill-scale trial results revealed that enzymatic prebleaching can be successfully employed with xylanases to reach the same bleach boosting efficacy. Laccase bleaching was also studied on hardwood pulp at a pH around 8.0, where most of the pulp mills in India are operating, in contrast to earlier studies on laccase enzyme bleaching, which were conducted at acidic pHs, i.e. 4.0 to 5.0. In case of laccase bleaching, interesting results were found wherein a bleach-boosting effect was observed even at pH 8.0. Further studies carried out with HOBT as mediator in comparison to the commonly used and expensive ABTS laccase mediator system (LMS resulted in improvement of the bleaching efficiency with reduction in demand of chlorine dioxide by more than 35%. Potential for further reduction was indicated by the brightness gain, when compared with a control using the DE(pD bleach sequence.

  7. CLONING, EXPRESSION, AND CHARACTERIZATION OF AN ALKALOPHILLIC ENDO-1,4-BETA-XYLANASE FROM PAENIBACILLUS SP. HPL-002

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    No-Joong Park,

    2011-12-01

    Full Text Available The biochemical properties of a purified enzyme of a new alkalophillic endo-1,4-beta-xylanase gene, KRICT PX2 (GU967374, which was isolated from Paenibacillus sp. HPL-002 (KCTC11410BP and expressed in E. coli, were investigated. The specific activity of the purified xylanase was 51.26 μmol/min/mg proteins. The Km and Vmax values of the protein for birch wood xylan were also verified to have 0.061 μM and 55.3 μmol/min/mg proteins, respectively. The optimum pH and temperature for the activity of the enzyme were pH 8~9 and 50oC, respectively, and, the activity was stably maintained at 40oC. Most metallic salts, ethylenediamine tetra-acetic acid, 2-mercaptoethanol, phenylmethane-sulphonyl fluoride, and furfural, have no impact on the enzyme’s activity at 1 mM. The simulated 3-D structure of this xylanase is similar to Xyn10B from Paenibacillus barcinonensis. Further research on the degradation of different-origin xylans and enzyme production will be necessary for practical applications.

  8. Effect of cuticular waxes compounds from table grapes on growth, germination and gene expression in Botrytis cinerea.

    Science.gov (United States)

    Silva-Moreno, Evelyn; Brito-Echeverría, Jocelyn; López, Miguel; Ríos, Juan; Balic, Iván; Campos-Vargas, Reinaldo; Polanco, Rubén

    2016-05-01

    Botrytis cinerea attacks a broad range of host causing significant economic losses in the worldwide fruit export industry. Hitherto, many studies have focused on the penetration mechanisms used by this phytopathogen, but little is known about the early stages of infection, especially those such as adhesion and germination. The aim of this work was to evaluate the effect of cuticular waxes compounds from table grapes on growth, germination and gene expression of B. cinerea. To accomplish this, growth was analyzed using as substrate n-alkanes extracted from waxes of fresh fruit (table grapes, blueberries and apricots). Subsequently, the main compounds of table grape waxes, oleanolic acid (OA) and n-fatty alcohols, were mixed to generate a matrix on which conidia of B. cinerea were added to assess their effect on germination and expression of bctub, bchtr and bchex genes. B. cinerea B05.10, isolated from grapes, increased its growth on a matrix composed by table grapes n-alkanes in comparison to a matrix made with n-alkanes from apricot or blueberries. Moreover, at 2.5 h, B05.10 germination increased 17 and 33 % in presence of n-alkanes from table grape, in comparison to conditions without alkanes or with blueberries alkanes, respectively. Finally, expression of bchtr and bchex showed a significant increase during the first hour after contact with n-fatty alcohols and OA. In conclusion, B. cinerea displays selectivity towards certain compounds found in host waxes, mainly n-fatty alcohols, which could be a good candidate to control this phytopathogen in early stages of infection.

  9. Priming for JA-dependent defenses using hexanoic acid is an effective mechanism to protect Arabidopsis against B. cinerea.

    Science.gov (United States)

    Kravchuk, Zhana; Vicedo, Begonya; Flors, Víctor; Camañes, Gemma; González-Bosch, Carmen; García-Agustín, Pilar

    2011-03-01

    Soil drench treatments with hexanoic acid can effectively protect Arabidopsis plants against Botrytis cinerea through a mechanism based on a stronger and faster accumulation of JA-dependent defenses. Plants impaired in ethylene, salicylic acid, abscisic acid or glutathion pathways showed intact protection by hexanoic acid upon B. cinerea infection. Accordingly, no significant changes in the SA marker gene PR-1 in either the SA or ABA hormone balance were observed in the infected and treated plants. In contrast, the JA signaling pathway showed dramatic changes after hexanoic acid treatment, mainly when the pathogen was present. The impaired JA mutants, jin1-2 and jar1, were unable to display hexanoic acid priming against the necrotroph. In addition, hexanoic acid-treated plants infected with B. cinerea showed priming in the expression of the PDF1.2, PR-4 and VSP1 genes implicated in the JA pathways. Moreover, JA and OPDA levels were primed at early stages by hexanoic acid. Treatments also stimulated increased callose accumulation in response to the pathogen. Although callose accumulation has proved an effective IR mechanism against B. cinerea, it is apparently not essential to express hexanoic acid-induced resistance (HxAc-IR) because the mutant pmr4.1 (callose synthesis defective mutant) is protected by treatment. We recently described how hexanoic acid treatments can protect tomato plants against B. cinerea by stimulating ABA-dependent callose deposition and by priming OPDA and JA-Ile production. We clearly demonstrate here that Hx-IR is a dependent plant species, since this acid protects Arabidopsis plants against the same necrotroph by priming JA-dependent defenses without enhancing callose accumulation.

  10. Transcriptome analysis reveals genes commonly induced by Botrytis cinerea infection, cold, drought and oxidative stresses in Arabidopsis.

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    Arjun Sham

    Full Text Available Signaling pathways controlling biotic and abiotic stress responses may interact synergistically or antagonistically. To identify the similarities and differences among responses to diverse stresses, we analyzed previously published microarray data on the transcriptomic responses of Arabidopsis to infection with Botrytis cinerea (a biotic stress, and to cold, drought, and oxidative stresses (abiotic stresses. Our analyses showed that at early stages after B. cinerea inoculation, 1498 genes were up-regulated (B. cinerea up-regulated genes; BUGs and 1138 genes were down-regulated (B. cinerea down-regulated genes; BDGs. We showed a unique program of gene expression was activated in response each biotic and abiotic stress, but that some genes were similarly induced or repressed by all of the tested stresses. Of the identified BUGs, 25%, 6% and 12% were also induced by cold, drought and oxidative stress, respectively; whereas 33%, 7% and 5.5% of the BDGs were also down-regulated by the same abiotic stresses. Coexpression and protein-protein interaction network analyses revealed a dynamic range in the expression levels of genes encoding regulatory proteins. Analysis of gene expression in response to electrophilic oxylipins suggested that these compounds are involved in mediating responses to B. cinerea infection and abiotic stress through TGA transcription factors. Our results suggest an overlap among genes involved in the responses to biotic and abiotic stresses in Arabidopsis. Changes in the transcript levels of genes encoding components of the cyclopentenone signaling pathway in response to biotic and abiotic stresses suggest that the oxylipin signal transduction pathway plays a role in plant defense. Identifying genes that are commonly expressed in response to environmental stresses, and further analyzing the functions of their encoded products, will increase our understanding of the plant stress response. This information could identify targets

  11. Arabidopsis AtERF15 positively regulates immunity against Pseudomonas syringae pv. tomato DC3000 and Botrytis cinerea

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    Huijuan eZhang

    2015-09-01

    Full Text Available Upon pathogen infection, activation of immune response requires effective transcriptional reprogramming that regulates inducible expression of a large set of defense genes. A number of ethylene-responsive factor transcription factors have been shown to play critical roles in regulating immune responses in plants. In the present study, we explored the functions of Arabidopsis AtERF15 in immune responses against Pseudomonas syringae pv. tomato (Pst DC3000, a (hemibiotrophic bacterial pathogen, and Botrytis cinerea, a necrotrophic fungal pathogen. Expression of AtERF15 was induced by infection of Pst DC3000 and B. cinerea and by treatments with salicylic acid (SA and methyl jasmonate. Biochemical assays demonstrated that AtERF15 is a nucleus-localized transcription activator. The AtERF15-overexpressing (AtERF15-OE plants displayed enhanced resistance while the AtERF15-RNAi plants exhibited decreased resistance against Pst DC3000 and B. cinerea. Meanwhile, Pst DC3000- or B. cinerea-induced expression of defense genes was upregulated in AtERF15-OE plants but downregulated in AtERF15-RNAi plants, as compared to the expression in wild type plants. In response to infection with B. cinerea, the AtERF15-OE plants accumulated less reactive oxygen species (ROS while the AtERF15-RNAi plants accumulated more ROS. The flg22- and chitin-induced oxidative burst was abolished and expression levels of the pattern-triggered immunity-responsive genes AtFRK1 and AtWRKY53 were suppressed in AtER15-RNAi plants upon treatment with flg22 or chitin. Furthermore, SA-induced defense response was also partially impaired in the AtERF15-RNAi plants. These data demonstrate that AtERF15 is a positive regulator of multiple layers of the immune responses in Arabidopsis.

  12. Preservation of Bacillus pumilus PU4-2 xylanases by immobilization technique into pollard and cation addition

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    T Haryati

    2010-03-01

    Full Text Available Utilization of by-product from agriculture as alternative source of feedstuff has been widely practiced. However their usage is limited due to high fiber content and low nutrient digestibility. The use of specific hydrolizing enzymes, xylanases are gaining importance because of their wide application in various industrial sectors especially in bioconversion of hemicellulosic material. This experiment was done to evaluate the effect of cation addition and immobilization of enzyme into pollard on stability of B. pumilus xylanase. The enzyme extract was purified by precipitation with 75% ammonium sulphate. Four kinds of cation (Ca2+, Fe3+, Mg2+, Zn2+ were added to the purified enzyme, at concentration of 1m M and stored at 4 and 27˚C. For immobilization process, the optimum enzyme concentration that will be added to pollard has been evaluated by analysis of xylanase activity and their recovery. The specific activity of enzyme after precipitation increased 1.8 times, from 420.3 to 765.2 U/mg protein. All cations act as activator which relative activity become 130.6; 139.0; 103.8 and 163.5% respectively. Concentration of 0.5mM Ca2+ and Fe3+ were most able to keep xylanases activity stable at 4˚C. The optimum composition of enzymes and pollard was 1.5 ml for 5 gram of pollard with recovery of xylanases activity of 82.2%. In immobilized enzyme, the activity of enzyme without cation addition is higher than that with addition of Ca2+ and Fe3+. Activity of enzyme stored at 4˚C is more stable than that at 27˚C. Immobilized enzyme is more stable for storage, which lasted for 7 weeks at 27˚C and 12 weeks at 4˚C compared to liquid enzyme which lasted for only 7 days at 27˚C and 13 days at 4˚C.

  13. Life cycle of Arcyria cinerea%灰团网菌的生活史

    Institute of Scientific and Technical Information of China (English)

    史立平; 李玉

    2012-01-01

    The life history of myxomycetes is of importance to study diversity of nutritive modes and phylogenesis of these organisms. Until now the reports concered were few. Moist chamber culture method,oat-agar culture method and scanning electron microscopy were used to study the process of the individual development of Arcyria cinerea. The life cycle of spore-to-spore were completed in agar culture in petridish. The result shows that the life cycle of Arcyria cinerea comprises a unicellular amoeboid or swarm cell stage,a multinucleate plasmodium stage and a sporulation stage. The spore of Arcyria cinerea is globular and the spore surface is spiny. The spore germinates by means of a poriform open and releases a single myxamoeba. The myxamoeba can move by amoeboid motion. When free water is available, myxamoebae can transform into swarm cells and swim in the water. Zygote forms into plasmodium. The mature plasmodium is white. The plasmodial type is the phaneroplasmodium which appears as a fan-shaped network of veins. Spores developed on agar are fertile and resemble those on natural substrate.%采用基物培养、燕麦-琼脂培养技术及扫描电镜技术对灰团网菌的个体发育过程进行了研究,在燕麦琼脂培养基上完成了从孢子到孢子的生活史.结果表明:灰团网菌生活史包括单核的黏变形体或游动胞、多核的营养体原质团以及孢子形成阶段.孢子球形,表面具疣突;孢子萌发为孔式,一个孢子可释放一个黏变形体.黏变形体常行变形运动,在有水的条件下,可转变为游动胞并具有游动的特性.合子形成原质团.成熟原质团白色,类型为显型,具有扇形网络状菌脉.琼脂培养基上获得的灰团网菌孢子与野生型相似,并具有可育性.

  14. Molecular characterization of boscalid resistance in field isolates of Botrytis cinerea from apple.

    Science.gov (United States)

    Yin, Y N; Kim, Y K; Xiao, C L

    2011-08-01

    Botrytis cinerea isolates obtained from apple orchards were screened for resistance to boscalid. Boscalid-resistant (BosR) isolates were classified into four phenotypes based on the levels of the concentration that inhibited fungal growth by 50% relative to control. Of the 220 isolates tested, 42 were resistant to boscalid, with resistant phenotypes ranging from low to very high resistance. There was cross resistance between boscalid and carboxin. Analysis of partial sequences of the iron-sulfur subunit of succinate dehydrogenase gene in B. cinerea (BcSdhB) from 13 BosR and 9 boscalid-sensitive (BosS) isolates showed that point mutations in BcSdhB leading to amino acid substitutions at the codon position 272 from histidine to either tyrosine (H272Y) or arginine (H272R) were correlated with boscalid resistance. Allele-specific polymerase chain reaction (PCR) analysis of 66 BosR isolates (including 24 additional isolates obtained from decayed apple fruit) showed that 19 carried the point mutation H272Y and 46 had the point mutation H272R, but 1 BosR isolate gave no amplification product. Analysis of the BcSdhB sequence of this isolate revealed a different point mutation at codon 225, resulting in a substitution of proline (P) by phenylalanine (F) (P225F). The results indicated that H272R/Y in BcSdhB were the dominant genotypes of mutants in field BosR isolates from apple. A multiplex allele-specific PCR assay was developed to detect point mutations H272R/Y in a single PCR amplification. Levels of boscalid resistance ranged from low to very high within isolates carrying either the H272R or H272Y mutation, indicating that, among BosR isolates, different BosR phenotypes (levels of resistance) were not associated with particular types of point mutations (H272R versus H272Y) in BcSdhB. Analysis of genetic relationships between 39 BosR and 56 BosS isolates based on three microsatellite markers showed that 39 BosR isolates and 30 BosS isolates were clustered into two groups

  15. Temperature-dependent growth of Botrytis cinerea isolates from potted plants.

    Science.gov (United States)

    Martínez, J A; Gómez-Bellot, M J; Bañón, S

    2009-01-01

    Botrytis cinereo is a common aggressive saprophyte fungus which also invades injured plant tissues, causing Botrytis blight (Grey mould) in many ornamental plants, including potted flowering plants. Several B. cinerea isolates from potted plants (Pelargonium x hortorum, Lantana camara, Lonicera japonica, Hydrangea macrophylla, and Cyclamen persicum) affected by Botrytis blight in the south of Spain were studied and identified by PCR. The isolates showed phenotypic differences between them, as previously reported by the authors. In this work we demonstrate that these isolates show different temperature-dependent growth phenomena, expressed as mycelial growth rates, conidiation (measured as the number of conidia per colony and time of appearance), mass of both aerial and submerged mycelia, and sclerotia production. Growth rates were assessed from differences in colony area and mass of both aerial and submerged mycelium growing in potato dextrose agar culture medium (PDA). Three temperatures were used to measure these variables (6, 16, and 26 degrees C) and to establish the differences among isolates by modelling the effects of temperature on the growth variables. B. cinerea showed a high degree of phenotypic variability and differences in its growth kinetics, depending on temperature and isolate in question. The isolate from P. x hortorum showed the greatest conidiation although this process did not depend on the temperatures assayed. The growth rate of the isolates from P. x hortorum was the highest. The growth rates in all the isolates were determined and the growth kinetics could be fitted to a typical equation of fungi growing on solid culture medium. The isolate from P. x hortorum was the most vigorous, while the least vigorous was the isolate from L. japonica. A relationship between mycelial growth rate, conidiation and aerial mycelium could be established. A temperature of 26 degrees C accelerated sclerotia production, but only in the isolate from C. persicum

  16. Fungicide resistance profiling in Botrytis cinerea populations from blueberries in California and Washington and their impact on control of gray mold

    Science.gov (United States)

    Gray mold caused by Botrytis cinerea is a major postharvest disease of blueberries grown in the Central Valley of California (CA) and western Washington State (WA). Sensitivities to boscalid, cyprodinil, fenhexamid, fludioxonil, and pyraclostrobin, representing five different fungicide classes, were...

  17. Assay Methods and Unit Definitions of Xylanase Activity%木聚糖酶酶活性测定方法及酶活性单位定义

    Institute of Scientific and Technical Information of China (English)

    王晓丹; 郭丽琼; 赵力超; 林俊芳

    2009-01-01

    The definition and determination of xylanase activity was unified in this article. Through summarizing and comparing the present knowledge on different procedure for the determination of xylanase activity,one unit of xylanase activity was defined as the quantity of enzyme required to liberate 1 (xmol of reducing sugar ( as xylose) per minute at the measure conditions . This DNS method of expressing xylanase activity is scientifically sound and we suggest that this method be utilized as the universal and standard method of determining xylanase activity.%为统一木聚糖酶酶活性的单位定义及测定方法进行了实验.通过对现有酶活性的单位定义及测定方法的比较分析,得出木聚糖酶的酶活性单位定义为:在特定条件下,每分钟水解木聚糖形成1μmol木糖(还原糖)所需酶量为1个酶活力单位(U),并且采用还原糖法中的DNS法作为测定木聚糖酶酶活性的方法,该法具有合理性和科学性,建议以此作为木聚糖酶酶活性测定及酶活性单位定义的统一方法.

  18. Research Progress on the Production of Xylanase by Microorganism Fermentation%木聚糖酶微生物发酵生产的研究进展

    Institute of Scientific and Technical Information of China (English)

    孙孝梅; 黄建忠

    2012-01-01

    Xylanase is one of the most important enzymes for industry application, which has a wide range of applica- tions, such as pulp and paper industry, food, feed, medicine and biotransformation. Microorganism fermentation is the major approach to produce Xylanase. This paper reviews the research progress on the aspects such as the breeding of high-yielding bacterial strain for Xylanase and the production of Xylanase by microorganism fermentation, and finally looks into the distance to the research directions and prospects on the production of Xylanase by microorganism fermentation.%木聚糖酶是一种重要的工业用酶制剂,广泛应用于造纸、食品、饲料、医药及生物转化等领域。木聚糖酶最主要的来源是通过微生物发酵而成。该文综述了木聚糖酶高产菌株的选育和微生物发酵生产等方面的研究进展,并展望了木聚糖酶微生物发酵生产的研究方向及前景。

  19. OPTIMIZATION OF CELLULASE-FREE XYLANASE PRODUCED BY A POTENTIAL THERMOALKALOPHILIC PAENIBACILLUS SP.N1 ISOLATED FROM HOT SPRINGS OF NORTHERN HIMALAYAS IN INDIA

    Directory of Open Access Journals (Sweden)

    Sanjeev Kumar Verma

    2012-08-01

    Full Text Available Hot spring bacteria are found a novel source of highly active xylanase enzyme with significant activity at high temperature. Among bacteria, Paenibacillus sp.N1 isolated from hot water spring of Manikaran, H.P., India showed highest 24.60 IU.ml-1 of cellulase-free xylanase on Reese medium. Growth conditions including medium, incubation time, pH, temperature, inoculum size, aminoacids, carbon sources, nitrogen sources and additives that affect the xylanase production by Paenibacillus sp.N1 were studied sequentially using the classical “change-one factor at a time” method. The optimal cultivation conditions predicated from canonical analysis of this model were achieved by using basal salt medium on 3rd day, pH 9.0, temperature 50ºC with inoculum size of 12.5%, phenylalanine as aminoacid, xylose as carbon source, (NH42HPO4 as nitrogen source and Tween 20 as detergent added with an approximate yield of 52.30 IU.ml-1 escalating the over level of xylanase production by 113.38%. A rare combination of all characters i.e. thermoalkalophilic nature and high units of cellulase-free xylanase produced from a new Paenibacillus sp.N1 make it of special industrial interest.

  20. A highly thermostable alkaline cellulase-free xylanase from thermoalkalophilic Bacillus sp. JB 99 suitable for paper and pulp industry: purification and characterization.

    Science.gov (United States)

    Shrinivas, Dengeti; Savitha, Gunashekaran; Raviranjan, Kumar; Naik, Gajanan Ramchandra

    2010-11-01

    A highly thermostable alkaline xylanase was purified to homogeneity from culture supernatant of Bacillus sp. JB 99 using DEAE-Sepharose and Sephadex G-100 gel filtration with 25.7-fold increase in activity and 43.5% recovery. The molecular weight of the purified xylanase was found to be 20 kDA by SDS-PAGE and zymogram analysis. The enzyme was optimally active at 70 °C, pH 8.0 and stable over pH range of 6.0-10.0.The relative activity at 9.0 and 10.0 were 90% and 85% of that of pH 8.0, respectively. The enzyme showed high thermal stability at 60 °C with 95% of its activity after 5 h. The K (m) and V (max) of enzyme for oat spelt xylan were 4.8 mg/ml and 218.6 µM min(-1) mg(-1), respectively. Analysis of N-terminal amino acid sequence revealed that the xylanase belongs to glycosyl hydrolase family 11 from thermoalkalophilic Bacillus sp. with basic pI. Substrate specificity showed a high activity on xylan-containing substrate and cellulase-free nature. The hydrolyzed product pattern of oat spelt xylan on thin-layer chromatography suggested xylanase as an endoxylanase. Due to these properties, xylanase from Bacillus sp. JB 99 was found to be highly compatible for paper and pulp industry.

  1. Phytochemical analysis of ethanolic extract of Dichrostachys Cinerea W and Arn leaves by a thin layer chromatography, high performance thin layer chromatography and column chromatography

    OpenAIRE

    Vijayalakshmi, M; K Periyanayagam; Kavitha, K; K Akilandeshwari

    2013-01-01

    Background: The leaves of Dichrostachys cinerea are used as laxative, diuretic, painkiller. It is also used in the treatment of gonorrhoea, boils, oedema, gout, veneral diseases and nasopharyngeal affections, etc. Materials and Methods: The Phytochemical investigation of ethanolic extract of D. cinerea leaves were performed by standard chemical tests, thin layer chromatography (TLC) by using various solvent systems, and by high performance liquid chromatography (HPTLC). Two compounds were...

  2. Oviposition preference and larval performance of Epiphyas postvittana (Lepidoptera: Tortricidae) on Botrytis cinerea (Helotiales: Sclerotiniaceae) infected berries of Vitis vinifera (Vitales: Vitaceae).

    Science.gov (United States)

    Rizvi, Syed Z M; Raman, Anantanarayanan; Wheatley, Warwick M; Cook, Geoffrey

    2016-04-01

    In this paper we tested the behavior of gravid Epiphyas postvittana in selecting the most-appropriate site for oviposition thus benefitting offspring performance. Our hypothesis was built on Jaenike's preference-performance hypothesis (also referred to as the "mother-knows-the-best" hypothesis). To test this, we used the interacting Epiphyas postvittana, its host Vitis vinifera, and the pathogenic microbe Botrytis cinerea system. Populations of E. postvittana and B. cinerea often exist concurrently on V. vinifera in Australasia and their interaction and mutual influence are currently being explored, although the suggestion presently is that the relationship between E. postvittana and B. cinerea is mutualistic. We tested the effect of volatiles from B. cinerea-infected berries and uninfected (control) berries of V. vinifera on the oviposition behavior of E. postvittana. We also characterized the effects of B. cinerea infection on the berries of V. vinifera on the growth and development of E. postvittana. Contrary to the preference-performance hypothesis, oviposition choices made by gravid E. postvittana did not result in the best offspring survival, development, and performance. The preference for oviposition by E. postvittana was strongly influenced by the olfactory and tactile cues. She laid fewer eggs on B. cinerea-infected berries compared to uninfected berries of V. vinifera. The larvae of E. postvittana showed no preference to uninfected berries of V. vinifera. The larvae fed on B. cinerea-infected berries of V. vinifera showing greater survival rate, shorter time to pupation, greater pupal mass, and on becoming adults they laid more numbers of eggs than the larvae that were enabled to feed on uninfected berries. The larvae of E. postvittana transport the conidia of B. cinerea and transmit grey-mould disease to uninfected berries of V. vinifera.

  3. Solidago canadensis L. Essential Oil Vapor Effectively Inhibits Botrytis cinerea Growth and Preserves Postharvest Quality of Strawberry as a Food Model System

    OpenAIRE

    Shumin Liu; Xingfeng Shao; Yanzhen Wei; Yonghua Li; Feng Xu; Hongfei Wang

    2016-01-01

    This study investigated the anti-fungal properties of Solidago canadensis L essential oil (SCLEO) against Botrytis cinerea in vitro, and its ability to control gray mold and maintain quality in strawberry fruits. SCLEO exhibited dose-dependent antifungal activity against B. cinerea and profoundly altered mycelial morphology, cellular ultrastructure, and membrane permeability as evaluated by scanning electron microscopy, transmission electron microscopy, and fluorescence microscopy. SCLEO vapo...

  4. Resistance evaluation of Chinese wild Vitis genotypes against Botrytis cinerea and different responses of resistant and susceptible hosts to the infection

    Directory of Open Access Journals (Sweden)

    Ran eWan

    2015-10-01

    Full Text Available The necrotrophic fungus Botrytis cinerea is a major threat to grapevine cultivation worldwide. A screen of 41 Vitis genotypes for leaf resistance to B. cinerea suggested species independent variation and revealed eighteen resistant Chinese wild Vitis genotypes, while most investigated V. vinifera, or its hybrids, were susceptible. A particularly resistant Chinese wild Vitis, ‘Pingli-5’ (V. sp. [Qinling grape] and a very susceptible V. vinifera cultivar, ‘Red Globe’ were selected for further study. Microscopic analysis demonstrated that B. cinerea growth was limitted during early infection on ‘Pingli-5’ before 24 hours post inoculation (hpi but not on Red Globe. It was found that reactive oxygen species (ROS and antioxidative system were associated with fungal growth. O2- accumulated similarly in B. cinerea 4 hpi on both Vitis genotypes. Lower levels of O2- (not H2O2 were detected 4 hpi and ROS (H2O2 and O2- accumulation from 8 hpi onwards was also lower in ‘Pingli-5’ leaves than in ‘Red Globe’ leaves. B. cinerea triggered sustained ROS production in ‘Red Globe’ but not in ‘Pingli-5’ with subsequent infection progresses. Red Globe displayed little change in antioxidative activities in response to B. cinerea infection, instead, antioxidative activities were highly and timely elevated in resistant ‘Pingli-5’ which correlated with its minimal ROS increases and its high resistance. These findings not only enhance our understanding of the resistance of Chinese wild Vitis species to B. cinerea, but also lay the foundation for breeding B. cinerea resistant grapes in the future.

  5. Continuous xylanase production with Aspergillus nidulans under pyridoxine limitation using a trickle bed reactor.

    Science.gov (United States)

    Müller, Michael; Prade, Rolf A; Segato, Fernando; Atiyeh, Hasan K; Wilkins, Mark R

    2015-01-01

    A trickle bed reactor (TBR) with recycle was designed and tested using Aspergillus nidulans with a pyridoxine marker and over-expressing/secreting recombinant client xylanase B (XynB). The pyridoxine marker prevented the fungus from synthesizing its own pyridoxine and fungus was unable to grow when no pyridoxine was present in the medium; however, enzyme production was unaffected. Uncontrolled mycelia growth that led to clogging of the TBR was observed when fungus without a pyridoxine marker was used for XynB production. Using the fungus with pyridoxine marker, the TBR was operated continuously for 18 days and achieved a XynB output of 41 U/ml with an influent and effluent flow rate of 0.5 ml/min and a recycle flow rate of 56 ml/min. Production yields in the TBR were 1.4 times greater than a static tray culture and between 1.1 and 67 times greater than yields for SSF enzyme production stated in the literature.

  6. High-yield recombinant xylanase production by Aspergillus nidulans under pyridoxine limitation.

    Science.gov (United States)

    Müller, Michael; Segato, Fernando; Prade, Rolf A; Wilkins, Mark R

    2014-10-01

    The present study investigated the limitation of pyridoxine on an Aspergillus nidulans culture that produces xylanase B (XynB) as a client enzyme and was unable to synthesize pyridoxine. This technique was used to limit cell growth and divert substrate to product formation for a surface grown culture that could be used in trickle bed reactors. It was observed that growth was limited when pyridoxine was absent, while enzyme production was unaffected. Enzyme production was 1,026 U after 480 h of continuous fermentation, which was similar to a culture that grew on medium with pyridoxine. Furthermore, the present study investigated the growth rate of A. nidulans with pyridoxine in the medium and determined the productivity of XynB production with and without pyridoxine. A maximum growth rate of 0.311/h was observed. The maximum XynB productivity of 21.14 U/g h was achieved when pyridoxine was not added to the medium.

  7. Ethanol/Water Pulps From Sugar Cane Straw and Their Biobleaching With Xylanase from Bacillus pumilus

    Science.gov (United States)

    Moriya, Regina Y.; Gonçalves, Adilson R.; Duarte, Marta C. T.

    The influence of independent variables (temperature and time) on the cooking of sugar cane straw with ethanol/water mixtures was studied to determine operating conditions that obtain pulp with high cellulose contents and a low lignin content. An experimental 22 design was applied for temperatures of 185 and 215°C, and time of 1 and 2.5 h with the ethanol/water mixture concentration and constant straw-to-solvent ratio. The system was scaled-up at 200°C cooking temperature for 2 h with 50% ethanol-water concentration, and 1∶10 (w/v) straw-to-solvent ratio to obtain a pulp with 3.14 cP viscosity, 58.09 kappa-number, and the chemical composition of the pulps were 3.2% pentosan and 31.5% lignin. Xylanase from Bacillus pumilus was then applied at a loading of 5-150 IU/g dry pulp in the sugar cane straw ethanol/water pulp at 50°C for 2 and 20 h. To ethanol/water pulps, the best enzyme dosage was found to be 20 IU/g dry pulp at 20 h, and a high enzyme dosage of 150 IU/g dry pulp did not decrease the kappa-number of the pulp.

  8. Ethanol from a biorefinery waste stream: Saccharification of amylase, protease and xylanase treated wheat bran.

    Science.gov (United States)

    Wood, Ian P; Cook, Nicola M; Wilson, David R; Ryden, Peter; Robertson, James A; Waldron, Keith W

    2016-05-01

    Biorefining aims to exploit the full value of plant material by sequentially extracting and valorising its components. Many studies focus on the saccharification of virgin biomass sources, but it may be more efficient to pre-extract high-value components before hydrolysis to fermentable sugars. In the current study, a bran residue from de-starched, protein depleted and xylanase treated wheat bran has been subjected to hydrothermal pretreatment, saccharification and fermentation procedures to convert the residue to ethanol. The most effective pretreatment conditions (>190 °C, 10 min) and saccharification conditions were identified following bench-scale liquid hot water pretreatment. Pre-extraction of enzymatically-hydrolysable starch and xylan reduced the release of furfural production, particularly when lower pretreatment severities were used. Pilot-scale steam explosion of the lignocellulosic residue followed by cellulase treatment and conversion to ethanol at a high substrate concentration (19%) gave an ethanol titre of ≈ 25 g/L or a yield of 93% of the theoretical maximum.

  9. Biosynthesis, purification and characterization of endoglucanase from a xylanase producing strain Aspergillus niger B03

    Directory of Open Access Journals (Sweden)

    Georgi Todorov Dobrev

    2012-03-01

    Full Text Available An extracellular endoglucanase was isolated from the culture liquid of xylanase producing strain Aspergillus niger B03. The enzyme was purified to a homogenous form, using consecutive ultrafiltration, anion exchange chromatography, and gel filtration. Endoglucanase was a monomer protein with a molecular weight of 26,900 Da determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and 28,800 Da determined by gel filtration. The optimal pH and temperature values for the enzyme action were 3.5 and 65ºC respectively. Endoglucanase was stable at 40ºC, pH 3.0 for 210 min. The substrate specificity of the enzyme was determined with carboxymethyl cellulose, filter paper, and different glycosides. Endoglucanase displayed maximum activity in the case of carboxymethyl cellulose, with a Km value of 21.01 mg/mL. The substrate specificity and the pattern of substrate degradation suggested that the enzyme is an endoglucanase. Endoglucanase showed a synergism with endoxylanase in corn cobs hydrolysis.

  10. SIDEROPHORE PRODUCING Pseudomonas AS PATHOGENIC Rhisoctonia solani AND Botrytis cinerea ANTAGONISTS

    Directory of Open Access Journals (Sweden)

    Martha Páez

    2005-06-01

    Full Text Available Pseudomonas aeruginosa, Pseudomonas putida biovar B, Pseudomonas marginalis y Burkholderia cepacia, aisladas de rizosfera y filosfera de plantas de rosa y alstroemeria, identificadas por ensayos bioquímicos y cultivadas en medio King B, mostraron propiedades antagónicas contra los patógenos (se usó medio PDA agar par el cultivo Rhizoctonia solani y Botrytis cinerea. Estas propiedades coincidieron con la presencia de un sideróforo, sustancia polar con bandas de absorción en 260 nm y 402 nm. Se observó incremento del crecimiento longitudinal de las plantas, medido sobre el tallo central, por influencia de P. putida biovar B, P. aeruginosa y P. marginalis. El crecimiento de rizomas (a: 0.05 fue notorio bajo la influencia de P. marginalis.

  11. Preharvest L-arginine treatment induced postharvest disease resistance to Botrysis cinerea in tomato fruits.

    Science.gov (United States)

    Zheng, Yang; Sheng, Jiping; Zhao, Ruirui; Zhang, Jian; Lv, Shengnan; Liu, Lingyi; Shen, Lin

    2011-06-22

    L-arginine is the precursor of nitric oxide (NO). In order to examine the influence of L-arginine on tomato fruit resistance, preharvest green mature tomato fruits (Solanum lycopersicum cv. No. 4 Zhongshu) were treated with 0.5, 1, and 5 mM L-arginine. The reduced lesion size (in diameter) on fruit caused by Botrytis cinerea, as well as activities of phenylalanine ammonia-lyase (PAL), Chitinase (CHI), β-1,3-glucanase (GLU), and polyphenoloxidase (PPO), was compared between L-arginine treated fruits and untreated fruits. We found that induced resistance increased and reached the highest level at 3-6 days after treatment. Endogenous NO concentrations were positively correlated with PAL, PPO, CHI, and GLU activities after treatment with Pearson coefficients of 0.71, 0.94, 0.97, and 0.87, respectively. These results indicate that arginine induces disease resistance via its effects on NO biosynthesis and defensive enzyme activity.

  12. ANTIDIARRHOEL ACTIVITY OF METHANOLIC EXTRACT OF VERNONIA CINEREA (L. LESS ON FEMALE ALBINO RATS

    Directory of Open Access Journals (Sweden)

    Panday Ganesh

    2011-05-01

    Full Text Available The present study was conducted with the objectives of investigating antidiarrhoel activity of Vernonia cinerea whole plant (Family-Compositae, collected from tarai region of Uttarakhand. The plant extracts were obtained via cold extraction method. For the purpose of evaluating antidiarrhoel efficacy of methanolic extract of the plant, rats were used as test animal. The time of onset of first wet faeces increased significantly and dose dependently by the extract. It was excellent at higher doses (100 & 200 mg/kg body wt., orally. It indicated reduction in peristaltic movement of gastro intestinal tract of animals. The antidiarrhoel activity was further confirmed by its significant and dose dependent decrease in number of wet faeces and number of total faeces in comparison to rats used as control.

  13. Efficacy of the Biofungicide PolyversumTM in Controlling Botrytis cinerea Pers. on Raspberry Fruits

    Directory of Open Access Journals (Sweden)

    Nenad Filajdić

    2006-01-01

    Full Text Available Efficacy of a biological fungicide product, Polyversum™ (Pythium oligandrum Drechsler, in controlling grey mould of raspberries Botrytis cinerea Pers. was tested at sites around Valjevo, Požega and Šabac in 2004, 2005 and 2006. The experiments were conducted anddata processed using EPPO methods.Efficacy was found to be significantly lower statistically in the experimental variants involving Polyversum™ biofungicide (E = 18.0%-53.9% than the standard botricide Ronilan-DF (E = 56.6%-90.3%.Our results show that the biological product Polyversum™ achieved significant efficacy although poorer than the standard fungicide Ronilan-DF. As it is a biological product almost entirely free of any toxicological and ecotoxicological limitations, it offers a significant advantage for widespread usage in plant protection.

  14. Botrytis pseudocinerea Is a Significant Pathogen of Several Crop Plants but Susceptible to Displacement by Fungicide-Resistant B. cinerea Strains.

    Science.gov (United States)

    Plesken, Cecilia; Weber, Roland W S; Rupp, Sabrina; Leroch, Michaela; Hahn, Matthias

    2015-10-01

    Botrytis cinerea is one of the most important pathogens worldwide, causing gray mold on a large variety of crops. Botrytis pseudocinerea has been found previously to occur together with B. cinerea in low abundance in vineyards and strawberry fields. Here, we report B. pseudocinerea to be common and sometimes dominant over B. cinerea on several fruit and vegetable crops in Germany. On apples with calyx end rot and on oilseed rape, it was the major gray mold species. Abundance of B. pseudocinerea was often negatively correlated with fungicide treatments. On cultivated strawberries, it was frequently found in spring but was largely displaced by B. cinerea following fungicide applications. Whereas B. cinerea strains with multiple-fungicide resistance were common in these fields, B. pseudocinerea almost never developed resistance to any fungicide even though resistance mutations occurred at similar frequencies in both species under laboratory conditions. The absence of resistance to quinone outside inhibitors in B. pseudocinerea was correlated with an intron in cytB preventing the major G143A resistance mutation. Our work indicates that B. pseudocinerea has a wide host range similar to that of B. cinerea and that it can become an important gray mold pathogen on cultivated plants.

  15. Tomato SlRbohB, a member of the NADPH oxidase family, is required for disease resistance against Botrytis cinerea and tolerance to drought stress

    Directory of Open Access Journals (Sweden)

    Xiaohui eLi

    2015-06-01

    Full Text Available NADPH oxidases (also known as respiratory burst oxidase homologues, Rbohs are the enzymes that catalyze the generation of reactive oxygen species (ROS in plants. In the present study, eight SlRboh genes were identified in tomato and their possible involvement in resistance to Botrytis cinerea and drought tolerance was examined. Expression of SlRbohs was induced by B. cinerea and Pseudomonas syringae pv. tomato but displayed distinct patterns. Virus-induced gene silencing (VIGS-based silencing of SlRbohB resulted in reduced resistance to B. cinerea but silencing of each of other SlRbohs did not affect the resistance. The SlRbohB-silenced plants accumulated more ROS and attenuated expression of defense genes after infection of B. cinerea than the nonsilenced plants. Silencing of SlRbohB also suppressed flg22-induced ROS burst and the expression of SlLrr22, a marker gene related to PAMP-triggered immunity (PTI. Transient expression of SlRbohB in Nicotiana benthamiana led to enhanced resistance to B. cinerea. Furthermore, silencing of SlRbohB resulted in decreased drought tolerance, accelerated water loss in leaves and altered expression of drought-responsive genes. Our data demonstrate that SlRbohB positively regulates the resistance to B. cinerea, flg22-induced PTI and drought tolerance in tomato.

  16. Dichrostachys cinerea and Acacia nilotica fruits as dry season feed supplements for goats in a semi-arid environment: Summary of a DFID funded project in Zimbabwe

    Energy Technology Data Exchange (ETDEWEB)

    Smith, T. [School of Agriculture, Policy and Development, University of Reading, Earley Gate, Reading (United Kingdom)]. E-mail: timsmith2@btopenworld.com; Mlambo, V. [Faculty of Agriculture, University of Swaziland, P.O. Luvengo (Swaziland); Sikosana, J.L.N.; Maphosa, V. [Department of Agricultural Research and Extension, Matopos Research Station, Bulawayo (Zimbabwe); Mueller-Harvey, I.; Owen, E. [School of Agriculture, Policy and Development, University of Reading, Earley Gate, Reading (United Kingdom)

    2005-08-19

    Indehiscent fruits of six tree species, common in Matabeleland were examined in in vitro and in vivo trials. The results for two of them, Acacia nilotica and Dichrostachys cinerea are presented here. Acacia nilotica contained more total phenolics than D. cinerea, but less nitrogen (N) and fibre (ADF and NDF). After 48 h incubation, in vitro OMD of both species was increased by PEG and NaOH or wood ash treatment, except when NaOH or wood ash were used in combination with PEG with D. cinerea fruits. DM intake, DMD were lowest and N-retention negative in goats fed A. nilotica as supplement. However when fed a supplement of D. cinerea, untreated or treated with PEG or NaOH, digestibility and N-retention were highest, and similar to a commercial goat meal, with the untreated fruit. In a trial in which milking does were supplemented with D. cinerea fruits, for 65 before and 65 days after kidding, kid birthweight and weaning weight were increased by supplementation. Deaths of twin-born kids were lowest in the supplemented but unmilked group. Supplementation with D. cinerea fruit resulted in improved goat performance. The only treatment applied of practical significance, wood ash, is currently being tested in an in vivo study. More research is required on detoxification of tannins, especially with A. nilotica. (author)

  17. Wounding induces local resistance but systemic susceptibility to Botrytis cinerea in pepper plants.

    Science.gov (United States)

    García, Tania; Gutiérrez, Jorge; Veloso, Javier; Gago-Fuentes, Raquel; Díaz, José

    2015-03-15

    Cotyledon wounding in pepper caused the early generation of hydrogen peroxide both locally (cotyledons) and systemically (upper true leaves). However, 72 h later there is a different wound response between local and systemic organs, as shown by resistance to the pathogenic fungus Botrytis cinerea, that increased locally and decreased systemically. Signaling by ethylene and jasmonic acid was assessed by using two inhibitors: 1-methylcyclopropene (MCP, inhibitor of ethylene receptors) and ibuprofen (inhibitor of jasmonate biosynthesis). MCP did not affect the modulation of resistance levels to Botrytis by wounding, ruling out the involvement of ethylene signaling. Ibuprofen did not inhibit wound-induced resistance at the local level, but inhibited wound-induced systemic susceptibility. Moreover, changes of biochemical and structural defenses in response to wounding were studied. Peroxidase activity and the expression of a peroxidase gene (CAPO1) increased locally as a response to wounding, but no changes were observed systemically. Lignin deposition was induced in wounded cotyledons, but was repressed in systemic leaves of wounded plants, whereas soluble phenolics did not change locally and decreased systemically. The expression of two other genes involved in plant defense (CABPR1 and CASC1) was also differentially regulated locally and systemically, pointing to a generalized increase in plant defenses at the local level and a systemic decrease as a response to wounding. Wound-induced defenses at the local level coincided with resistance to the necrotroph fungus B. cinerea, whereas depleted defenses in systemic leaves of wounded plants correlated to induced susceptibility against this pathogen. It may be that the local response acts as a sink of energy resources to mount a defense against pathogens, whereas in systemic organs the resources for defense are lower.

  18. Growth Simulation and Discrimination of Botrytis cinerea, Rhizopus stolonifer and Colletotrichum acutatum Using Hyperspectral Reflectance Imaging.

    Science.gov (United States)

    Sun, Ye; Gu, Xinzhe; Wang, Zhenjie; Huang, Yangmin; Wei, Yingying; Zhang, Miaomiao; Tu, Kang; Pan, Leiqing

    2015-01-01

    This research aimed to develop a rapid and nondestructive method to model the growth and discrimination of spoilage fungi, like Botrytis cinerea, Rhizopus stolonifer and Colletotrichum acutatum, based on hyperspectral imaging system (HIS). A hyperspectral imaging system was used to measure the spectral response of fungi inoculated on potato dextrose agar plates and stored at 28°C and 85% RH. The fungi were analyzed every 12 h over two days during growth, and optimal simulation models were built based on HIS parameters. The results showed that the coefficients of determination (R2) of simulation models for testing datasets were 0.7223 to 0.9914, and the sum square error (SSE) and root mean square error (RMSE) were in a range of 2.03-53.40×10(-4) and 0.011-0.756, respectively. The correlation coefficients between the HIS parameters and colony forming units of fungi were high from 0.887 to 0.957. In addition, fungi species was discriminated by partial least squares discrimination analysis (PLSDA), with the classification accuracy of 97.5% for the test dataset at 36 h. The application of this method in real food has been addressed through the analysis of Botrytis cinerea, Rhizopus stolonifer and Colletotrichum acutatum inoculated in peaches, demonstrating that the HIS technique was effective for simulation of fungal infection in real food. This paper supplied a new technique and useful information for further study into modeling the growth of fungi and detecting fruit spoilage caused by fungi based on HIS.

  19. Growth Simulation and Discrimination of Botrytis cinerea, Rhizopus stolonifer and Colletotrichum acutatum Using Hyperspectral Reflectance Imaging.

    Directory of Open Access Journals (Sweden)

    Ye Sun

    Full Text Available This research aimed to develop a rapid and nondestructive method to model the growth and discrimination of spoilage fungi, like Botrytis cinerea, Rhizopus stolonifer and Colletotrichum acutatum, based on hyperspectral imaging system (HIS. A hyperspectral imaging system was used to measure the spectral response of fungi inoculated on potato dextrose agar plates and stored at 28°C and 85% RH. The fungi were analyzed every 12 h over two days during growth, and optimal simulation models were built based on HIS parameters. The results showed that the coefficients of determination (R2 of simulation models for testing datasets were 0.7223 to 0.9914, and the sum square error (SSE and root mean square error (RMSE were in a range of 2.03-53.40×10(-4 and 0.011-0.756, respectively. The correlation coefficients between the HIS parameters and colony forming units of fungi were high from 0.887 to 0.957. In addition, fungi species was discriminated by partial least squares discrimination analysis (PLSDA, with the classification accuracy of 97.5% for the test dataset at 36 h. The application of this method in real food has been addressed through the analysis of Botrytis cinerea, Rhizopus stolonifer and Colletotrichum acutatum inoculated in peaches, demonstrating that the HIS technique was effective for simulation of fungal infection in real food. This paper supplied a new technique and useful information for further study into modeling the growth of fungi and detecting fruit spoilage caused by fungi based on HIS.

  20. Arabidopsis Elongator subunit 2 positively contributes to resistance to the necrotrophic fungal pathogens Botrytis cinerea and Alternaria brassicicola.

    Science.gov (United States)

    Wang, Chenggang; Ding, Yezhang; Yao, Jin; Zhang, Yanping; Sun, Yijun; Colee, James; Mou, Zhonglin

    2015-09-01

    The evolutionarily conserved Elongator complex functions in diverse biological processes including salicylic acid-mediated immune response. However, how Elongator functions in jasmonic acid (JA)/ethylene (ET)-mediated defense is unknown. Here, we show that Elongator is required for full induction of the JA/ET defense pathway marker gene PLANT DEFENSIN1.2 (PDF1.2) and for resistance to the necrotrophic fungal pathogens Botrytis cinerea and Alternaria brassicicola. A loss-of-function mutation in the Arabidopsis Elongator subunit 2 (ELP2) alters B. cinerea-induced transcriptome reprogramming. Interestingly, in elp2, expression of WRKY33, OCTADECANOID-RESPONSIVE ARABIDOPSIS AP2/ERF59 (ORA59), and PDF1.2 is inhibited, whereas transcription of MYC2 and its target genes is enhanced. However, overexpression of WRKY33 or ORA59 and mutation of MYC2 fail to restore PDF1.2 expression and B. cinerea resistance in elp2, suggesting that ELP2 is required for induction of not only WRKY33 and ORA59 but also PDF1.2. Moreover, elp2 is as susceptible as coronatine-insensitive1 (coi1) and ethylene-insensitive2 (ein2) to B. cinerea, indicating that ELP2 is an important player in B. cinerea resistance. Further analysis of the lesion sizes on the double mutants elp2 coi1 and elp2 ein2 and the corresponding single mutants revealed that the function of ELP2 overlaps with COI1 and is additive to EIN2 for B. cinerea resistance. Finally, basal histone acetylation levels in the coding regions of WRKY33, ORA59, and PDF1.2 are reduced in elp2 and a functional ELP2-GFP fusion protein binds to the chromatin of these genes, suggesting that constitutive ELP2-mediated histone acetylation may be required for full activation of the WRKY33/ORA59/PDF1.2 transcriptional cascade.

  1. Exogenous dietary xylanase ameliorates viscosity-induced anti-nutritional effects in wheat-based diets for White Pekin ducks (Anas platyrinchos domesticus).

    Science.gov (United States)

    Adeola, Olayiwola; Bedford, Michael R

    2004-07-01

    Nutrient utilisation and growth performance responses of White Pekin ducks (Anas platyrinchos domesticus) offered diets containing low- or high-viscosity wheat supplemented with xylanase were investigated in two studies. In Expt 1, six diets consisting of low-viscosity wheat or high-viscosity wheat supplemented with 0.0, 1.5 or 3.0 g xylanase (2590 units/g)/kg diet were used in a true metabolisable energy (TME) bioassay with eight 8-week-old ducks per diet group. In Expt 2, eight pens of ten 3-d-old ducks per pen for each of six wheat-based diets arranged in a 2 x 3 factorial of low-viscosity or high-viscosity wheat and 0.0, 1.5 or 3.0 g xylanase/kg were used in a 42 d growth study. High-viscosity wheat depressed (Pducks.

  2. Phylogenetic analysis of β-xylanase SRXL1 of Sporisorium reilianum and its relationship with families (GH10 and GH11) of Ascomycetes and Basidiomycetes.

    Science.gov (United States)

    Álvarez-Cervantes, Jorge; Díaz-Godínez, Gerardo; Mercado-Flores, Yuridia; Gupta, Vijai Kumar; Anducho-Reyes, Miguel Angel

    2016-04-04

    In this paper, the amino acid sequence of the β-xylanase SRXL1 of Sporisorium reilianum, which is a pathogenic fungus of maize was used as a model protein to find its phylogenetic relationship with other xylanases of Ascomycetes and Basidiomycetes and the information obtained allowed to establish a hypothesis of monophyly and of biological role. 84 amino acid sequences of β-xylanase obtained from the GenBank database was used. Groupings analysis of higher-level in the Pfam database allowed to determine that the proteins under study were classified into the GH10 and GH11 families, based on the regions of highly conserved amino acids, 233-318 and 180-193 respectively, where glutamate residues are responsible for the catalysis.

  3. Aspects microbiologiques de la production par fermentation solide des endo-beta-1,4-xylanases de moisissures : le cas de Penicillium canescens

    Directory of Open Access Journals (Sweden)

    Assamoi AA.

    2009-01-01

    Full Text Available Microbial aspects of endo-β-1,4-xylanase production in solid-state fermentation by Penicillia: the case of Penicillium canescens. Production of xylanases by Penicillium canescens 10-10c is the research object in Walloon Center of Industrial Biology. Previous works used submerged or liquid fermentation. The actual works are oriented more and more towards solid fermentation from agricultural or agro-alimentary residues. In addition to the valorization of these residues, solid-state fermentation reaches an increasingly significant interest in various other fields like the biological breakdown of the solid residues, the bioremediation of the organic pollutants in the grounds and the reduction of the air pollution by the biofiltration. Xylanase is an industrial enzyme used in general in extraction and clarification processes. P. canescens can produce an activity of it, particularly in its balanced forms of xylanases, beta-xylosidase and arabinosidase, and not contaminated by cellulolytic and amylolytic activities. It is a hyper producing strain of xylanase. The production rate is one of the highest in literature (535 U.ml-1 and 9,632 U.g-1 in Erlenmeyer flasks, in submerged and solid state fermentation, respectively. The biobleaching activity of the cellulose pulp by the purified enzyme is higher than a commercial preparation of xylanases from Trichoderma longibrachiatum used industrially. It has a complete hydrolysis degree of 40% (on glucuronoxylan and 35% (on arabinoxylan at 55°C and at pH of 5.9. These characteristics lead to many industrial applications of this enzyme. That is why the optimization of its production by the solid-state fermentation at the laboratory scale in order to define a policy for the industrial transposition later is carried out. This article presents a summary of the scientific literature on this subject.

  4. Co-cultivation of mutant Penicillium oxalicum SAU(E)-3.510 and Pleurotus ostreatus for simultaneous biosynthesis of xylanase and laccase under solid-state fermentation.

    Science.gov (United States)

    Dwivedi, Pallavi; Vivekanand, V; Pareek, Nidhi; Sharma, Amit; Singh, Rajesh P

    2011-10-01

    Co-cultivation of mutant Penicillium oxalicum SAU(E)-3.510 and Pleurotus ostreatus MTCC 1804 was evaluated for the production of xylanase-laccase mixture under solid-state fermentation (SSF) condition. Growth compatibility between mutant P. oxalicum SAU(E)-3.510 and white rot fungi (P. ostreatus MTCC 1804, Trametes hirsuta MTCC 136 and Pycnoporus sp. MTCC 137) was analyzed by growing them on potato dextrose agar plate. Extracellular enzyme activities were determined spectrophotometrically. Under derived conditions, paired culturing of mutant P. oxalicum SAU(E)-3.510 and P. ostreatus MTCC 1804 resulted in 58% and 33% higher levels of xylanase and laccase production, respectively. A combination of sugarcane bagasse and black gram husk in a ratio of 3:1 was found to be the most ideal solid substrate and support for fungal colonization and enzyme production during co-cultivation. Maximum levels of xylanase (8205.31 ± 168.31 IU g(-1)) and laccase (375.53 ± 34.17 IU g(-1)) during SSF were obtained by using 4 g of solid support with 80% of moisture content. Furthermore, expressions of both xylanase and laccase were characterized during mixed culture by zymogram analysis. Improved levels of xylanase and laccase biosynthesis were achieved by co-culturing the mutant P. oxalicum SAU(E)-3.510 and P. ostreatus MTCC 1804. This may be because of efficient substrate utilization as compared to their respective monocultures in the presence of lignin degradation compounds because of synergistic action of xylanase and laccase. Understanding and developing the process of co-cultivation appears productive for the development of mixed enzyme preparation with tremendous potential for biobleaching.

  5. Critical aspects of analysis of Micrococcus luteus, Neisseria cinerea, and Pseudomonas fluorescens by means of capillary electrophoresis.

    Science.gov (United States)

    Hoerr, Verena; Stich, August; Holzgrabe, Ulrike

    2004-10-01

    Within the frame of our study we investigated Microccocus luteus, Neisseria cinerea, and Pseudomonas fluorescens by means of capillary zone electrophoresis (CZE). They form chains and clusters on a different scale, which can be reflected in the electropherograms. A low buffer concentration of Tris-borate and Na2EDTA containing a polymeric matrix of 0.0125% poly(ethylene) oxide (PEO) was used. Key factors were the standardization and optimization of CE conditions, buffer solution, and pretreatment of bacterial samples, which are not transferable to different bacterial strains, in general. The different compositions of the cell wall of on the one hand Gram-positive (M. luteus) and Gram-negative (N. cinerea) cocci and on the other hand Gram-negative, rod-shaped bacteria (P. fluorescens), are probably responsible for the different pretreatment conditions.

  6. Chemical constituents in the essential oil of the endemic plantCotula cinerea (Del.) from the southwest of Algeria

    Institute of Scientific and Technical Information of China (English)

    Mohammed Djellouli; Houcine Benmehdi; Siham Mammeri; Abdellah Moussaoui; Laid Ziane; Noureddine Hamidi

    2015-01-01

    Objective:To extract and identify the main constituents of the essential oil ofCotula cinerea (Del.) (Asteraceae family) from southwest of Algeria. Methods: The essential oils obtained by hydrodistillation, from the aerial parts of the endemic plantCotula cinerea which was collected in the region of Sahara fromsouthwest of Algeria, were analyzed by gas chromatography-mass spectrometry. Results: A total of 33 compounds were identified representing 98.66% of the oil. The main compounds were (E)-citral (24.01%), limonene epoxide cis- (18.26%), thymol methyl ether (15.04%), carvacrol (15.03%), trans-carveol (13.79%), carvone (3.06%) and trans-piperitol (2.54%). Conclusions: The main constituents in essential oil of the aerial part of the plant from southwest of Algeria were different from that collected from southeast of Algeria or in Morocco.

  7. Chemical constituents in the essential oil of the endemic plant Cotula cinerea (Del.) from the southwest of Algeria简

    Institute of Scientific and Technical Information of China (English)

    Mohammed; Djellouli; Houcine; Benmehdi; Siham; Mammeri; Abdellah; Moussaoui; Laid; Ziane; Noureddine; Hamidi

    2015-01-01

    Objective: To extract and identify the main constituents of the essential oil of Cotula cinerea(Del.)(Asteraceae family) from southwest of Algeria.Methods: The essential oils obtained by hydrodistillation, from the aerial parts of the endemic plant Cotula cinerea which was collected in the region of Sahara from southwest of Algeria, were analyzed by gas chromatography-mass spectrometry.Results: A total of 33 compounds were identified representing 98.66% of the oil. The main compounds were(E)-citral(24.01%), limonene epoxide cis-(18.26%), thymol methyl ether(15.04%), carvacrol(15.03%), trans-carveol(13.79%), carvone(3.06%) and trans-piperitol(2.54%).Conclusions: The main constituents in essential oil of the aerial part of the plant from southwest of Algeria were different from that collected from southeast of Algeria or in Morocco.

  8. The p450 monooxygenase BcABA1 is essential for abscisic acid biosynthesis in Botrytis cinerea

    DEFF Research Database (Denmark)

    Siewers, V.; Smedsgaard, Jørn; Tudzynski, P.

    2004-01-01

    The phytopathogenic ascomycete Botrytis cinerea is known to produce abscisic acid (ABA), which is thought to be involved in host-pathogen interaction. Biochemical analyses had previously shown that, in contrast to higher plants, the fungal ABA biosynthesis probably does not proceed via carotenoids...... but involves direct cyclization of farnesyl diphosphate and subsequent oxidation steps. We present here evidence that this "direct" pathway is indeed the only one used by an ABA-overproducing strain of B. cinerea. Targeted inactivation of the gene bccpr1 encoding a cytochrome P450 oxidoreductase reduced...... the ABA production significantly, proving the involvement of P450 monooxygenases in the pathway. Expression analysis of 28 different putative P450 monooxygenase genes revealed two that were induced under ABA biosynthesis conditions. Targeted inactivation showed that one of these, bcaba1, is essential...

  9. Cellulose and hemicellulose-degrading enzymes in Fusarium commune transcriptome and functional characterization of three identified xylanases.

    Science.gov (United States)

    Huang, Yuhong; Busk, Peter Kamp; Lange, Lene

    2015-06-01

    Specific enzymes from plant-pathogenic microbes demonstrate high effectiveness for natural lignocellulosic biomass degradation and utilization. The secreted lignocellulolytic enzymes of Fusarium species have not been investigated comprehensively, however. In this study we compared cellulose and hemicellulose-degrading enzymes of classical fungal enzyme producers with those of Fusarium species. The results indicated that Fusarium species are robust cellulose and hemicellulose degraders. Wheat bran, carboxymethylcellulose and xylan-based growth media induced a broad spectrum of lignocellulolytic enzymes in Fusarium commune. Prediction of the cellulose and hemicellulose-degrading enzymes in the F. commune transcriptome using peptide pattern recognition revealed 147 genes encoding glycoside hydrolases and six genes encoding lytic polysaccharide monooxygenases (AA9 and AA11), including all relevant cellulose decomposing enzymes (GH3, GH5, GH6, GH7, GH9, GH45 and AA9), and abundant hemicellulases. We further applied peptide pattern recognition to reveal nine and seven subfamilies of GH10 and GH11 family enzymes, respectively. The uncharacterized XYL10A, XYL10B and XYL11 enzymes of F. commune were classified, respectively, into GH10 subfamily 1, subfamily 3 and GH11 subfamily 1. These xylanases were successfully expressed in the PichiaPink™ system with the following properties: the purified recombinant XYL10A had interesting high specific activity; XYL10B was active at alkaline conditions with both endo-1,4-β-d-xylanase and β-xylosidase activities; and XYL11 was a true xylanase characterized by high substrate specificity. These results indicate that F. commune with genetic modification is a promising source of enzymes for the decomposition of lignocellulosic biomass.

  10. Simultaneous bioethanol distillery wastewater treatment and xylanase production by the phyllosphere yeast Pseudozyma antarctica GB-4(0).

    Science.gov (United States)

    Watanabe, Takashi; Suzuki, Ken; Sato, Ikuo; Morita, Tomotake; Koike, Hideaki; Shinozaki, Yukiko; Ueda, Hirokazu; Koitabashi, Motoo; Kitamoto, Hiroko K

    2015-12-01

    Bioethanol production using lignocellulosic biomass generates lignocellulosic bioethanol distillery wastewater (LBDW) that contains a large amount of xylose, making it a potential inexpensive source of xylose for biomaterials production. The main goal of this study was the production of useful enzymes from LBDW during treatment of this wastewater. In this study, we found that xylose strongly induced two yeast strains, Pseudozyma antarctica T-34 and GB-4(0), to produce novel xylanases, PaXynT and PaXynG, respectively. The nucleotide sequence of PaXynT [accession No. DF196774 (GAC73192.1)], obtained from the genome database of strain T-34 using its N-terminal amino acid sequence, was 91% identical to that of PaXynG (accession No. AB901085), and the deduced amino acid sequence is 98% identical. The specific activities of the purified PaXynT and PaXynG were about 52 U/mg. The optimal pH and temperature for both enzymes' activities were 5.2 and 50°C, respectively. They hydrolyzed xylan to xylose and neither had β-xylosidase (EC 3.2.1.37) activity, indicating that they are endo-β-xylanases (EC 3.2.1.8). With these results, we expect that PaXyns can be employed in saccharizing lignocellulosic biomass materials for the production of useful products just like other endoxylanases. After 72 h of LBDW fed-batch cultivation using a jar-fermentor, strain GB-4(0) produced 17.3 U/ml (corresponding to about 0.3 g/l) of PaXynG and removed 63% of dissolved organic carbon and 87% of dissolved total phosphorus from LBDW. These results demonstrate the potential of P. antarctica for xylanase production during LBDW treatment.

  11. Conformation analysis of a surface loop that controls active site access in the GH11 xylanase A from Bacillus subtilis.

    Science.gov (United States)

    Vieira, Davi Serradella; Ward, Richard John

    2012-04-01

    Xylanases (EC 3.2.1.8 endo-1,4-glycosyl hydrolase) catalyze the hydrolysis of xylan, an abundant hemicellulose of plant cell walls. Access to the catalytic site of GH11 xylanases is regulated by movement of a short β-hairpin, the so-called thumb region, which can adopt open or closed conformations. A crystallographic study has shown that the D11F/R122D mutant of the GH11 xylanase A from Bacillus subtilis (BsXA) displays a stable "open" conformation, and here we report a molecular dynamics simulation study comparing this mutant with the native enzyme over a range of temperatures. The mutant open conformation was stable at 300 and 328 K, however it showed a transition to the closed state at 338 K. Analysis of dihedral angles identified thumb region residues Y113 and T123 as key hinge points which determine the open-closed transition at 338 K. Although the D11F/R122D mutations result in a reduction in local inter-intramolecular hydrogen bonding, the global energies of the open and closed conformations in the native enzyme are equivalent, suggesting that the two conformations are equally accessible. These results indicate that the thumb region shows a broader degree of energetically permissible conformations which regulate the access to the active site region. The R122D mutation contributes to the stability of the open conformation, but is not essential for thumb dynamics, i.e., the wild type enzyme can also adapt to the open conformation.

  12. Xylanase and Protease Increase Solubilization of Non-Starch Polysaccharides and Nutrient Release of Corn- and Wheat Distillers Dried Grains with Solubles

    DEFF Research Database (Denmark)

    Pedersen, Mads Brøgger; Dalsgaard, Søren; Arent, Susan

    2015-01-01

    The use of distiller dried grains with solubles (DDGS) as alternative to conventional animal feed for non-ruminants is challenged by the high content of non-starch polysaccharides and varying protein quality. In this study the enzymatic degradation of corn- and wheat DDGS was evaluated, in vitro......, by use of four xylanases from two different glycoside hydrolase families, GH10 and GH11, along with protease and phytase. Wheat DDGS showed the highest degree of enzymatic degradation due to a lower degree of cell wall complexity compared with that of corn DDGS. For corn DDGS, the combination of xylanase...

  13. The enhancement of enzymatic hydrolysis of lignocellulosic substrates by the addition of accessory enzymes such as xylanase: is it an additive or synergistic effect?

    Directory of Open Access Journals (Sweden)

    Saddler Jack N

    2011-10-01

    Full Text Available Abstract Background We and other workers have shown that accessory enzymes, such as β-glucosidase, xylanase, and cellulase cofactors, such as GH61, can considerably enhance the hydrolysis effectiveness of cellulase cocktails when added to pretreated lignocellulosic substrates. It is generally acknowledged that, among the several factors that hamper our current ability to attain efficient lignocellulosic biomass conversion yields at low enzyme loadings, a major problem lies in our incomplete understanding of the cooperative action of the different enzymes acting on pretreated lignocellulosic substrates. Results The reported work assessed the interaction between cellulase and xylanase enzymes and their potential to improve the hydrolysis efficiency of various pretreated lignocellulosic substrates when added at low protein loadings. When xylanases were added to the minimum amount of cellulase enzymes required to achieve 70% cellulose hydrolysis of steam pretreated corn stover (SPCS, or used to partially replace the equivalent cellulase dose, both approaches resulted in enhanced enzymatic hydrolysis. However, the xylanase supplementation approach increased the total protein loading required to achieve significant improvements in hydrolysis (an additive effect, whereas the partial replacement of cellulases with xylanase resulted in similar improvements in hydrolysis without increasing enzyme loading (a synergistic effect. The enhancement resulting from xylanase-aided synergism was higher when enzymes were added simultaneously at the beginning of hydrolysis. This co-hydrolysis of the xylan also influenced the gross fiber characteristics (for example, fiber swelling resulting in increased accessibility of the cellulose to the cellulase enzymes. These apparent increases in accessibility enhanced the steam pretreated corn stover digestibility, resulting in three times faster cellulose and xylan hydrolysis, a seven-fold decrease in cellulase loading and

  14. The WRKY57 Transcription Factor Affects the Expression of Jasmonate ZIM-Domain Genes Transcriptionally to Compromise Botrytis cinerea Resistance.

    Science.gov (United States)

    Jiang, Yanjuan; Yu, Diqiu

    2016-08-01

    Although necrotrophic pathogens cause many devastating plant diseases, our understanding of the plant defense response to them is limited. Here, we found that loss of function of WRKY57 enhanced the resistance of Arabidopsis (Arabidopsis thaliana) against Botrytis cinerea infection. Further investigation suggested that the negative regulation of WRKY57 against B cinerea depends on the jasmonic acid (JA) signaling pathway. Chromatin immunoprecipitation experiments revealed that WRKY57 directly binds to the promoters of JASMONATE ZIM-DOMAIN1 (JAZ1) and JAZ5, encoding two important repressors of the JA signaling pathway, and activates their transcription. In vivo and in vitro experiments demonstrated that WRKY57 interacts with nuclear-encoded SIGMA FACTOR BINDING PROTEIN1 (SIB1) and SIB2. Further experiments display that the same domain, the VQ motif, of SIB1 and SIB2 interact with WRKY33 and WRKY57. Moreover, transient transcriptional activity assays confirmed that WRKY57 and WRKY33 competitively regulate JAZ1 and JAZ5, SIB1 and SIB2 further enhance these competitions of WRKY57 to WRKY33. Therefore, coordinated regulation of Arabidopsis against B cinerea by transcription activators and repressors would benefit plants by allowing fine regulation of defense.

  15. Arabidopsis WRKY33 is a key transcriptional regulator of hormonal and metabolic responses toward Botrytis cinerea infection.

    Science.gov (United States)

    Birkenbihl, Rainer P; Diezel, Celia; Somssich, Imre E

    2012-05-01

    The Arabidopsis (Arabidopsis thaliana) transcription factor WRKY33 is essential for defense toward the necrotrophic fungus Botrytis cinerea. Here, we aimed at identifying early transcriptional responses mediated by WRKY33. Global expression profiling on susceptible wrky33 and resistant wild-type plants uncovered massive differential transcriptional reprogramming upon B. cinerea infection. Subsequent detailed kinetic analyses revealed that loss of WRKY33 function results in inappropriate activation of the salicylic acid (SA)-related host response and elevated SA levels post infection and in the down-regulation of jasmonic acid (JA)-associated responses at later stages. This down-regulation appears to involve direct activation of several jasmonate ZIM-domain genes, encoding repressors of the JA-response pathway, by loss of WRKY33 function and by additional SA-dependent WRKY factors. Moreover, genes involved in redox homeostasis, SA signaling, ethylene-JA-mediated cross-communication, and camalexin biosynthesis were identified as direct targets of WRKY33. Genetic studies indicate that although SA-mediated repression of the JA pathway may contribute to the susceptibility of wrky33 plants to B. cinerea, it is insufficient for WRKY33-mediated resistance. Thus, WRKY33 apparently directly targets other still unidentified components that are also critical for establishing full resistance toward this necrotroph.

  16. Characteristics of inositol phosphorylceramide synthase and effects of aureobasidin A on growth and pathogenicity of Botrytis cinerea.

    Science.gov (United States)

    Wang, Xin-hui; Guo, Xing-Jun; Li, Hong-Ye; Gou, Ping

    2015-01-01

    Inositol phosphorylceramide (IPC) synthase is the key enzyme with highly conserved sequences, which is involved in fungal sphingolipid biosynthesis. The antibiotic aureobasidin A (AbA) induces the death of fungi through inhibiting IPC synthase activity. The mutations of AUR1 gene coding IPC synthase in fungi and protozoa causes a resistance to AbA. However, the mechanism of AbA resistance is still elusive. In this paper, we generated two mutants of Botrytis cinerea with AbA-resistance, BcAUR1a and BcAUR1b, through UV irradiation. BcAUR1a lost an intron and BcAUR1b had three amino acid mutations (L197P, F288S and T323A) in the AUR1 gene. AbA strongly inhibits the activity of IPC synthase in wild-type B. cinerea, which leads to distinct changes in cell morphology, including the delay in conidial germination, excessive branching near the tip of the germ tube and mycelium, and the inhibition of the mycelium growth. Further, AbA prevents the infection of wild-type B. cinerea in tomato fruits via reducing oxalic acid secretion and the activity of cellulase and pectinase. On the contrary, AbA has no effect on the growth and pathogenicity of the two mutants. Although both mutants show a similar AbA resistance, the molecular mechanisms might be different between the two mutants.

  17. Genome-wide characterization of ISR induced in Arabidopsis thaliana by Trichoderma hamatum T382 against Botrytis cinerea infection

    Directory of Open Access Journals (Sweden)

    Janick eMathys

    2012-05-01

    Full Text Available In this study, the molecular basis of the induced systemic resistance (ISR in Arabidopsis thaliana by the biocontrol fungus Trichoderma hamatum T382 against the phytopathogen Botrytis cinerea B05-10 was unraveled by microarray analysis both before (ISR-prime and after (ISR-boost additional pathogen inoculation. The observed high numbers of differentially expressed genes allowed us to classify them according to the biological pathways in which they are involved. By focusing on pathways instead of genes, a holistic picture of the mechanisms underlying ISR emerged. In general, a close resemblance is observed between ISR-prime and systemic acquired resistance (SAR, the systemic defense response that is triggered in plants upon pathogen infection leading to increased resistance towards secondary infections. Treatment with Trichoderma hamatum T382 primes the plant (ISR-prime, resulting in an accelerated activation of the defense response against Botrytis cinerea during ISR-boost and a subsequent moderation of the Botrytis cinerea induced defense response (BIDR. Microarray results were confirmed for representative genes by qRT-PCR, by analysis of transgenic plants expressing relevant promoter-GUS constructs and by phenotypic analysis of mutants affected in various defense-related pathways, thereby proving the validity of our approach.

  18. Genome-Wide Characterization of ISR Induced in Arabidopsis thaliana by Trichoderma hamatum T382 Against Botrytis cinerea Infection.

    Science.gov (United States)

    Mathys, Janick; De Cremer, Kaat; Timmermans, Pieter; Van Kerckhove, Stefan; Lievens, Bart; Vanhaecke, Mieke; Cammue, Bruno P A; De Coninck, Barbara

    2012-01-01

    In this study, the molecular basis of the induced systemic resistance (ISR) in Arabidopsis thaliana by the biocontrol fungus Trichoderma hamatum T382 against the phytopathogen Botrytis cinerea B05-10 was unraveled by microarray analysis both before (ISR-prime) and after (ISR-boost) additional pathogen inoculation. The observed high numbers of differentially expressed genes allowed us to classify them according to the biological pathways in which they are involved. By focusing on pathways instead of genes, a holistic picture of the mechanisms underlying ISR emerged. In general, a close resemblance is observed between ISR-prime and systemic acquired resistance, the systemic defense response that is triggered in plants upon pathogen infection leading to increased resistance toward secondary infections. Treatment with T. hamatum T382 primes the plant (ISR-prime), resulting in an accelerated activation of the defense response against B. cinerea during ISR-boost and a subsequent moderation of the B. cinerea induced defense response. Microarray results were validated for representative genes by qRT-PCR. The involvement of various defense-related pathways was confirmed by phenotypic analysis of mutants affected in these pathways, thereby proving the validity of our approach. Combined with additional anthocyanin analysis data these results all point to the involvement of the phenylpropanoid pathway in T. hamatum T382-induced ISR.

  19. Origin of (-)-geosmin on grapes: on the complementary action of two fungi, botrytis cinerea and penicillium expansum.

    Science.gov (United States)

    La Guerche, Stéphane; Chamont, Sophie; Blancard, Dominique; Dubourdieu, Denis; Darriet, Philippe

    2005-08-01

    One of the consequences of rot on grapes is the development of volatile compounds giving fungal, mouldy or earthy odours. Among these compounds, (-)-geosmin (trans-1,10-dimethyl-trans-9-decalol), a powerful aromatic compound with an earthy smell is a persistent defect in grape juice and wines made with at least partially rotten grapes. A microbiota analysis of rotten grapes containing (-)-geosmin was carried out on sites from four French regions from 1999 to 2002, to clarify the involvement in geosmin appearance of Streptomyces spp. and Penicillium spp., two types of microorganisms present on grape, that are known for their ability to produce geosmin. In earthy grapes, Botrytis cinerea was largely present. Different species of Streptomyces were also isolated, but their pH sensitivity was an extremely limiting parameter for their development on grape juice, grapes or stem, and consequently for their potentiality to generate geosmin in the vineyard. Penicillium expansum, producing geosmin on a model medium, was omnipresent. Penicillium carneum, which is also a geosmin producer, was represented by a single colony during the 4 years of this study. P. expansum alone was able to produce geosmin on a model medium but not on grapes. However, after 7 days' pre-culture of some B. cinerea strains on grape juice, this juice became favourable to geosmin production by P. expansum. We demonstrated the necessary and complementary action of B. cinerea and P. expansum in geosmin production in grape juice and in crushed grape berries.

  20. Transcriptome analysis reveals regulatory networks underlying differential susceptibility to Botrytis cinerea in response to nitrogen availability in Solanum lycopersicum.

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    Andrea eVega

    2015-11-01

    Full Text Available Nitrogen (N is one of the main limiting nutrients for plant growth and crop yield. It is well documented that changes in nitrate availability, the main N source found in agricultural soils, influences a myriad of developmental programs and processes including the plant defense response. Indeed, many agronomical reports indicate that the plant N nutritional status influences their ability to respond effectively when challenged by different pathogens. However, the molecular mechanisms involved in N-modulation of plant susceptibility to pathogens are poorly characterized. In this work, we show that Solanum lycopersicum defense response to the necrotrophic fungus Botrytis cinerea is affected by plant N availability, with higher susceptibility in nitrate-limiting conditions. Global gene expression responses of tomato against B. cinerea under contrasting nitrate conditions reveals that plant primary metabolism is affected by the fungal infection regardless of N regimes. This result suggests that differential susceptibility to pathogen attack under contrasting N conditions is not only explained by a metabolic alteration. We used a systems biology approach to identify the transcriptional regulatory network implicated in plant response to the fungus infection under contrasting nitrate conditions. Interestingly, hub genes in this network are known key transcription factors involved in ethylene and jasmonic acid signaling. This result positions these hormones as key integrators of nitrate and defense against B. cinerea in tomato plants. Our results provide insights into potential crosstalk mechanisms between necrotrophic defense response and N status in plants.

  1. Effect of temperature on the morphological characteristics of Botrytis cinerea and its correlated with the genetic variability

    Institute of Scientific and Technical Information of China (English)

    Jorge G Fernndez; Martn A Fernndez-Baldo; Gabriela Sansone; Viviana Calvente; Delia Benuzzi; Eloy Salinas; Julio Raba; Mara I Sanz

    2014-01-01

    Objective: To study the effect of temperature on the morphological characteristics of Botrytiscinerea (B. cinerea) and its correlated with the genetic variability. B. cinerea is a plant-pathogenic fungus that produces the disease known as grey mould in a wide variety of agriculturally important hosts in many countries.Methods:Six strains from different host collected have been isolated and characterized by several methods as mycelial growth, fungicide resistance, pathogenicity and the effects of the temperature. Also was analyzed by PCR and distinguished by the presence or absence of transposable elements.Results:Results showed that clear morphological differences exist between strains at the temperature of 4, 12 and 28 °C. All strains analyzed molecularly were classified as Group II (transposa-type). Demonstrating a negative correlation between mycelial growth and other characteristics as the fungicide resistance and pathogenicity. Lastly, it is difficult to establish relationships phenotypic and genotypic between strains of B. cinerea.Conclusions:The results indicated that the mycelial growth, resistance at fungicide and pathogenicity are independent of the characteristics molecular, however, are dependent of a factor such as temperature.

  2. Genome-Wide Characterization of ISR Induced in Arabidopsis thaliana by Trichoderma hamatum T382 Against Botrytis cinerea Infection

    Science.gov (United States)

    Mathys, Janick; De Cremer, Kaat; Timmermans, Pieter; Van Kerckhove, Stefan; Lievens, Bart; Vanhaecke, Mieke; Cammue, Bruno P. A.; De Coninck, Barbara

    2012-01-01

    In this study, the molecular basis of the induced systemic resistance (ISR) in Arabidopsis thaliana by the biocontrol fungus Trichoderma hamatum T382 against the phytopathogen Botrytis cinerea B05-10 was unraveled by microarray analysis both before (ISR-prime) and after (ISR-boost) additional pathogen inoculation. The observed high numbers of differentially expressed genes allowed us to classify them according to the biological pathways in which they are involved. By focusing on pathways instead of genes, a holistic picture of the mechanisms underlying ISR emerged. In general, a close resemblance is observed between ISR-prime and systemic acquired resistance, the systemic defense response that is triggered in plants upon pathogen infection leading to increased resistance toward secondary infections. Treatment with T. hamatum T382 primes the plant (ISR-prime), resulting in an accelerated activation of the defense response against B. cinerea during ISR-boost and a subsequent moderation of the B. cinerea induced defense response. Microarray results were validated for representative genes by qRT-PCR. The involvement of various defense-related pathways was confirmed by phenotypic analysis of mutants affected in these pathways, thereby proving the validity of our approach. Combined with additional anthocyanin analysis data these results all point to the involvement of the phenylpropanoid pathway in T. hamatum T382-induced ISR. PMID:22661981

  3. Bacillus isolates from the spermosphere of peas and dwarf French beans with antifungal activity against Botrytis cinerea and Pythium species.

    Science.gov (United States)

    Walker, R; Powell, A A; Seddon, B

    1998-05-01

    A range of isolation procedures including washing, sonication and incubation in nutrient broth were used separately and in combination to obtain potential bacterial antagonists to Botrytis cinerea and Pythium mamillatum from the testae and cotyledons of peas and dwarf French beans. Heat treatment was also used to bias this selection towards spore-forming bacteria. Ninety-two bacterial isolates were obtained, 72 of which were provisionally characterized as species of Bacillus. Four of these Bacillus isolates (B3, C1, D4 and J7) displayed distinct antagonism in vitro against Botrytis cinerea and P. mamillatum when screened using dual culture analysis. Further characterization of these antagonists using API 50CHB biochemical profiling identified isolate D4 as Bacillus polymyxa and isolates B3, C1 and J7 as strains of B. subtilis. In vitro screening techniques, using cell-free and heat-killed extracts of liquid cultures against Botrytis cinerea, demonstrated the production of antifungal compounds by these four Bacillus antagonists. With each isolate the antifungal activity was found not to be either exclusively spore-bound nor released entirely into the medium but present in both fractions. The antifungal compounds produced by these isolates were shown to be heat-stable. Their identification, production and release require further study for exploitation as biocontrol systems.

  4. D-Xylose fermentation, xylitol production and xylanase activities by seven new species of Sugiyamaella.

    Science.gov (United States)

    Sena, Letícia M F; Morais, Camila G; Lopes, Mariana R; Santos, Renata O; Uetanabaro, Ana P T; Morais, Paula B; Vital, Marcos J S; de Morais, Marcos A; Lachance, Marc-André; Rosa, Carlos A

    2017-01-01

    Sixteen yeast isolates identified as belonging to the genus Sugiyamaella were studied in relation to D-xylose fermentation, xylitol production, and xylanase activities. The yeasts were recovered from rotting wood and sugarcane bagasse samples in different Brazilian regions. Sequence analyses of the internal transcribed spacer (ITS) region and the D1/D2 domains of large subunit rRNA gene showed that these isolates belong to seven new species. The species are described here as Sugiyamaella ayubii f.a., sp. nov. (UFMG-CM-Y607(T) = CBS 14108(T)), Sugiyamaella bahiana f.a., sp. nov. (UFMG-CM-Y304(T) = CBS 13474(T)), Sugiyamaella bonitensis f.a., sp. nov. (UFMG-CM-Y608(T) = CBS 14270(T)), Sugiyamaella carassensis f.a., sp. nov. (UFMG-CM-Y606(T) = CBS 14107(T)), Sugiyamaella ligni f.a., sp. nov. (UFMG-CM-Y295(T) = CBS 13482(T)), Sugiyamaella valenteae f.a., sp. nov. (UFMG-CM-Y609(T) = CBS 14109(T)) and Sugiyamaella xylolytica f.a., sp. nov. (UFMG-CM-Y348(T) = CBS 13493(T)). Strains of the described species S. boreocaroliniensis, S. lignohabitans, S. novakii and S. xylanicola, isolated from rotting wood of Brazilian ecosystems, were also compared for traits relevant to xylose metabolism. S. valenteae sp. nov., S. xylolytica sp. nov., S. bahiana sp. nov., S. bonitensis sp. nov., S. boreocarolinensis, S. lignohabitans and S. xylanicola were able to ferment D-xylose to ethanol. Xylitol production was observed for all Sugiyamaella species studied, except for S. ayubii sp. nov. All species studied showed xylanolytic activity, with S. xylanicola, S. lignohabitans and S. valenteae sp. nov. having the highest values. Our results suggest these Sugiyamaella species have good potential for biotechnological applications.

  5. Improvement of A Fungitoxicity Test of Fungicide to Botrytis cinerea%一种测定杀菌剂对灰霉病菌(Botrytis cinerea) 毒力方法的改进

    Institute of Scientific and Technical Information of China (English)

    陈治芳; 韩秀英; 王文桥; 赵建江; 张小风; 马志强; 刘颖超

    2011-01-01

    To seek for a high sensitive and high accuracy method to screen fungicides, in this research, a new test (cucumber cotyledons spray inoculation test) of Botrytis cinerea was established through inoculation on cucumber cotyledons with conidial suspension of B. cinerea amended with 1% yeast extract, 2.5% glucose and 10% carrot juice. The results showed that the test exhibits high disease incidence (100%) and uniform disease severity (>98%), good repetition and simple operation. Inhibitory activity of tebuconazole, mancozeb and iprodione to Botrytis cinerea was tested and compared by cucumber cotyledons spray inoculation test, mycelial growth rate test and conidial germination test, it was found that a high correlation was shown between the logarithmic concentrations of fungicide and probability values of inhibition rate, the correlation coefficients were all above 0.95. This method could be used in fungicide sensitivity test and screening resistance level of fungicide against Botrytis cinerea.%为探索一种灵敏度高、准确性高的筛选灰霉病防治药剂的试验方法,笔者通过在灰霉病菌(Botrytis cinerea)分生孢子悬浮液中加入1%酵母浸膏、2.5%葡萄糖和10%胡萝卜汁,接种离体黄瓜子叶,病叶率达100%,发病均匀度达98%以上,确立了一种重复性好、操作简便的测定杀菌剂对灰霉病菌毒力的新方法一黄瓜子叶喷雾法.以该方法、菌丝生长速率法和孢子萌发法分别测定比较了戊唑醇、代森锰锌和异菌脲对灰霉病菌的毒力,发现药剂浓度的对数值与其抑制率的几率值之间有很高的相关性,相关系数均大于0.95.该方法可用于灰霉病菌抗药性监测和杀菌剂敏感性测定.

  6. Pseudomonas spp.-induced systemic resistance to Botrytis cinerea is associated with induction and priming of defence responses in grapevine.

    Science.gov (United States)

    Verhagen, Bas W M; Trotel-Aziz, Patricia; Couderchet, Michel; Höfte, Monica; Aziz, Aziz

    2010-01-01

    Non-pathogenic rhizobacteria Pseudomonas spp. can reduce disease in plant tissues through induction of a defence state known as induced systemic resistance (ISR). This resistance is based on multiple bacterial determinants, but nothing is known about the mechanisms underlying rhizobacteria-induced resistance in grapevine. In this study, the ability of Pseudomonas fluorescens CHA0 and Pseudomonas aeruginosa 7NSK2 to induce resistance in grapevine against Botrytis cinerea is demonstrated. Both strains also triggered an oxidative burst and phytoalexin (i.e. resveratrol and viniferin) accumulation in grape cells and primed leaves for accelerated phytoalexin production upon challenge with B. cinerea. Treatment of cell cultures with crude cell extracts of bacteria strongly enhanced oxidative burst, but resulted in comparable amounts of phytoalexins and resistance to B. cinerea to those induced by living bacteria. This suggests the production of bacterial compounds serving as inducers of disease resistance. Using other strains with different characteristics, it is shown that P. fluorescens WCS417 (Pch-deficient), P. putida WCS358 (Pch- and SA-deficient) and P. fluorescens Q2-87 (a DAPG producer) were all capable of inducing resistance to an extent similar to that induced by CHA0. However, in response to WCS417 (Pch-negative) the amount of H2O2 induced is less than for the CHA0. WCS417 induced low phytoalexin levels in cells and lost the capacity to prime for phytoalexins in the leaves. This suggests that, depending on the strain, SA, pyochelin, and DAPG are potentially effective in inducing or priming defence responses. The 7NSK2 mutants, KMPCH (Pch- and Pvd-negative) and KMPCH-567 (Pch-, Pvd-, and SA-negative) induced only partial resistance to B. cinerea. However, the amount of H2O2 triggered by KMPCH and KMPCH-567 was similar to that induced by 7NSK2. Both mutants also led to a low level of phytoalexins in grapevine cells, while KMPCH slightly primed grapevine leaves

  7. Characterization of iprodione resistance in Botrytis cinerea from strawberry and blackberry.

    Science.gov (United States)

    Grabke, Anja; Fernández-Ortuño, Dolores; Amiri, Achour; Li, Xingpeng; Peres, Natália A; Smith, Powell; Schnabel, Guido

    2014-04-01

    Gray mold, caused by the fungal pathogen Botrytis cinerea, is one of the most destructive diseases of strawberry. Control of the disease in commercial fields is largely dependent on the application of fungicides, including the dicarboximide iprodione. Single-spore isolates were collected from strawberry fields in Florida, North Carolina, and South Carolina and subjected to an assay using conidial germination that distinguished sensitive (S) isolates from isolates with various levels of resistance to iprodione. Of the 245 isolates, 1 was highly resistant (HR), 5 were moderately resistant (MR), and 43 had low resistance (LR) to iprodione. LR and MR strains were found in the Florida population and in 9 of 11 locations from North Carolina and South Carolina, indicating that resistance was widespread but accounted for only a relatively small percentage of the B. cinerea population. Sequence analysis of the target gene bos1, which codes for a class III histidine kinase, revealed that the MR phenotype was associated with Q369P and N373S mutations and that the LR phenotype was associated with either a I365S or a I365N mutation. The I365S and I365N mutations were also present in five additionally included HR isolates from North Carolina and South Carolina blackberry fields and one HR isolate from a Virginia strawberry field but no mutation or mutation combinations in bos1 were uniquely associated with the HR phenotype. Expression analysis of bos1 in S and HR isolates did not reveal convincing evidence of the gene's involvement in HR resistance either. The six HR isolates had three different phenotypes with respect to their sensitivity to fludioxonil; two were S, two were LR, and two were MR. The fludioxonil LR and MR isolates were also resistant to tolnaftate, an indication of multidrug efflux pump activity. These data suggest that, in addition to point mutations in bos1, drug efflux pump activity and potentially a third mechanism of resistance may be contributing to the

  8. Independent Emergence of Resistance to Seven Chemical Classes of Fungicides in Botrytis cinerea.

    Science.gov (United States)

    Fernández-Ortuño, Dolores; Grabke, Anja; Li, Xingpeng; Schnabel, Guido

    2015-04-01

    Gray mold, caused by the fungal pathogen Botrytis cinerea, is one of the most destructive diseases of small fruit crops and control is largely dependent on the application of fungicides. As part of a region-wide resistance-monitoring program that investigated 1,890 B. cinerea isolates from 189 fields in 10 states of the United States, we identified seven isolates (0.4%) from five locations in four different states with unprecedented resistance to all seven Fungicide Resistance Action Committee (FRAC) codes with single-site modes of action including FRAC 1, 2, 7, 9, 11, 12, and 17 registered in the United States for gray mold control. Resistance to thiophanate-methyl, iprodione, boscalid, pyraclostrobin, and fenhexamid was based on target gene mutations that conferred E198A and F200Y in β-tubulin, I365N/S in Bos1, H272R/Y in SdhB, G143A in Cytb, and T63I and F412S in Erg27. Isolates were grouped into MDR1 and MDR1h phenotypes based on sensitivity to fludioxonil and variations in transcription factor mrr1. MDR1h isolates had a previously described 3-bp deletion at position 497 in mrr1. Expression of ABC transporter atrB was increased in MDR1 isolates but highest in MDR1h isolates. None of the isolates with seven single resistances (SR) had identical nucleotide variations in target genes, indicating that they emerged independently. Multifungicide resistance phenotypes did not exhibit significant fitness penalties for the parameters used in this study, but MDR1h isolates produced more sclerotia at low temperatures and exhibited increased sensitivity to salt stress. In this study we show that current resistance management strategies have not been able to prevent the geographically independent development of resistance to all seven site-specific fungicides currently registered for gray mold control in the United States and document the presence of MDR1h in North America.

  9. Engineering a high-performance, metagenomic-derived novel xylanase with improved soluble protein yield and thermostability.

    Science.gov (United States)

    Qian, Changli; Liu, Ning; Yan, Xing; Wang, Qian; Zhou, Zhihua; Wang, Qianfu

    2015-03-01

    The novel termite gut metagenomic-derived GH11 xylanase gene xyl7 was expressed in Escherichia coli BL21, and the purified XYL7 enzyme exhibited high specific activity (6340U/mg) and broad pH active range of 5.5-10.0. Directed evolution was employed to enhance the thermostability of XYL7; two mutants (XYL7-TC and XYL7-TS) showed a 250-fold increase in half-life at 55°C, with a 10°C increase in optimal temperature compared to that of wild-type XYL7. A truncated enzyme (XYL7-Tr3) acquired by protein engineering showed similar catalytic properties as the wild-type, with a tenfold increase in soluble protein yield by the mutant. The reducing sugar produced by XYL7-TC was about fourfold greater than that produced by their parents when incubated with xylan at 60°C for 4h. The engineered novel xylanase exhibited superior enzymatic performance and showed promise as an excellent candidate for industrial application due to its high specific activity, stability and soluble protein yield.

  10. Generation of transgenic wheat (Triticum aestivum L.) accumulating heterologous endo-xylanase or ferulic acid esterase in the endosperm

    Energy Technology Data Exchange (ETDEWEB)

    Harholt, Jesper; Bach, Inga C; Lind-Bouquin, Solveig; Nunan, Kylie J.; Madrid, Susan M.; Brinch-Pedersen, Henrik; Holm, Preben B.; Scheller, Henrik V.

    2009-12-08

    Endo-xylanase (from Bacillus subtilis) or ferulic acid esterase (from Aspergillus niger) were expressed in wheat under the control of the endosperm specific 1DX5 glutenin promoter. Constructs both with and without the endoplasmic reticulum retention signal KDEL were used. Transgenic plants were recovered in all four cases but no qualitative differences could be observed whether KDEL was added or not. Endo-xylanase activity in transgenic grains was increased between two and three fold relative to wild type. The grains were shriveled and had a 25-33% decrease in mass. Extensive analysis of the cell walls showed a 10-15% increase in arabinose to xylose ratio, a 50% increase in the proportion of water extractable arabinoxylan, and a shift in the MW of the water extractable arabinoxylan from being mainly larger than 85 kD to being between 2 kD and 85 kD. Ferulic acid esterase expressing grains were also shriveled and the seed weight was decreased by 20-50%. No ferulic acid esterase activity could be detected in wild type grains whereas ferulic acid esterase activity was detected in transgenic lines. The grain cell walls had 15-40% increase in water unextractable arabinoxylan and a decrease in monomeric ferulic acid between 13 and 34%. In all the plants the observed changes are consistent with a plant response that serves to minimize the effect of the heterologously expressed enzymes by increasing arabinoxylan biosynthesis and cross-linking.

  11. Generation of transgenic wheat (Triticum aestivum L.) accumulating heterologous endo-xylanase or ferulic acid esterase in the endosperm.

    Science.gov (United States)

    Harholt, Jesper; Bach, Inga C; Lind-Bouquin, Solveig; Nunan, Kylie J; Madrid, Susan M; Brinch-Pedersen, Henrik; Holm, Preben B; Scheller, Henrik V

    2010-04-01

    Endo-xylanase (from Bacillus subtilis) or ferulic acid esterase (from Aspergillus niger) were expressed in wheat under the control of the endosperm-specific 1DX5 glutenin promoter. Constructs both with and without the endoplasmic reticulum retention signal (Lys-Asp-Glu-Leu) KDEL were used. Transgenic plants were recovered in all four cases but no qualitative differences could be observed whether KDEL was added or not. Endo-xylanase activity in transgenic grains was increased between two and threefold relative to wild type. The grains were shrivelled and had a 25%-33% decrease in mass. Extensive analysis of the cell walls showed a 10%-15% increase in arabinose to xylose ratio, a 50% increase in the proportion of water-extractable arabinoxylan, and a shift in the MW of the water-extractable arabinoxylan from being mainly larger than 85 kD to being between 2 and 85 kD. Ferulic acid esterase-expressing grains were also shrivelled, and the seed weight was decreased by 20%-50%. No ferulic acid esterase activity could be detected in wild-type grains whereas ferulic acid esterase activity was detected in transgenic lines. The grain cell walls had 15%-40% increase in water-unextractable arabinoxylan and a decrease in monomeric ferulic acid between 13% and 34%. In all the plants, the observed changes are consistent with a plant response that serves to minimize the effect of the heterologously expressed enzymes by increasing arabinoxylan biosynthesis and cross-linking.

  12. THE INFLUENCE OF ASOCIATED SUPPLEMENT OF ALFA AMYLASE AND XYLANASE ON THE RHEOLOGY OF DOUGH CONCEARNING ITS CONSTITOGRAPHICAL PARAMETERS

    Directory of Open Access Journals (Sweden)

    RODICA CHEREJI

    2013-07-01

    Full Text Available In this paper we determined the influence of associated supplement of alfa amylase and xylanase on the rheology of dough concearning its constitographical parameters : maximum pressure (Pr max, (mb and the absorbed water (Wa, %. The analysis on the consistograph were conducted for constant hydration at the consistency of 500 UF. Determinations were made on 4 types of flour and optimal dosages were found for each enzyme, after which we prepared the optimal dosage of the enzymes in the compund for flour F1 and F2 : P1-840000 U. SKB/100 kg flour +8100 U. FXU /100 kg flour, P2-840000 U. SKB/100 kg flour+16200 U. FXU, /100 kg flour , P3-840000 U. SKB/100 kg flour+24300 U. FXU/100 kg flour , and for F3 and F4 thus: P1-280000 U. SKB/100 kg flour +8100 U. FXU /100 kg flour, P2- 280000 U. SKB/100 kg flour+16200 U. FXU/100 kg flour, P3-280000 U. SKB/100 kg flour+24300 U. FXU/100 kg flour. Fungous α-amylase and xylanase were used in these concentrations to establish which one is more apropriate to be added in flour to obtain superior quality of bread: finer texture of the crumb, prolongation of the freashness of the bread, improvind the colour and flavour, emproving the slicing ability.

  13. Characterizing and improving the thermostability of purified xylanase from Aspergillus niger DFR-5 grown on solid-state-medium

    Directory of Open Access Journals (Sweden)

    Ajay Pal

    2010-12-01

    Full Text Available  The thermostability of absolutely purified xylanase from Aspergillus niger DFR-5 was improved using polyols. Supplementation of sorbitol at 2M concentration was found to increase the half-life and D-value of xylanase at elevated temperatures (45-70ºC. Thermodynamic parameters associated with the process were analyzed revealing that the stability at higher temperatures was due to the increased enthalpy (∆Hº and free energy (∆Gº change of enzyme denaturation in the presence of sorbitol. The negative values of ∆Sº (-150.093 Jmol-1K-1 at 70ºC clearly indicated that enzyme underwent a significant process of aggregation during denaturation. The enzyme required divalent cations for maximum activity and inhibited by chelator. The diminution of activity by various thiol-binding agents and enhancement by reducing agents like β-ME confirmed the essentiality of cysteine for catalysis. The enzyme had a half-life and D-value of 277 and 921 days when stored at 4 ºC.

  14. A xylanase gene directly cloned from the genomic DNA of alkaline wastewater sludge showing application potential in the paper industry.

    Science.gov (United States)

    Zhao, Yanyu; Luo, Huiying; Meng, Kun; Shi, Pengjun; Wang, Guozeng; Yang, Peilong; Yuan, Tiezheng; Yao, Bin

    2011-09-01

    A xylanase gene, aws-2x, was directly cloned from the genomic DNA of the alkaline wastewater sludge using degenerated PCR and modified TAIL-PCR. The deduced amino acid sequence of AWS-2x shared the highest identity (60%) with the xylanase from Chryseobacterium gleum belonging to the glycosyl hydrolase GH family 10. Recombinant AWS-2x was expressed in Escherichia coli BL21 (DE3) and purified to electrophoretic homogeneity. The enzyme showed maximal activity at pH 7.5 and 55 °C, maintained more than 50% of maximal activity when assayed at pH 9.0, and was stable over a wide pH range from 4.0 to 11.0. The specific activity of AWS-2x towards hardwood xylan (beechwood and birchwood xylan) was significantly higher than that to cereal xylan (oat spelt xylan and wheat arabinoxylan). These properties make AWS-2x a potential candidate for application in the pulp and paper industry.

  15. Engineering the hydrophobic residues of a GH11 xylanase impacts its adsorption onto lignin and its thermostability.

    Science.gov (United States)

    Rakotoarivonina, Harivony; Hermant, Béatrice; Aubry, Nathalie; Rémond, Caroline

    2015-12-01

    This study aimed to characterise the parameters governing the non-specific adsorption of a xylanase from Thermobacillus xylanilyticus (Tx-Xyn11) onto lignin isolated from maize stems. Such adsorption may be due to hydrophobic interactions between Tx-Xyn11 and lignin. Our strategy was to mutate hydrophobic residues present on the surface of Tx- Xyn11 into non-hydrophobic residues. Three mutants (P1, P2, and P3) with altered hydrophobic regions were produced and characterised. The thermostability of the P1 mutant was largely decreased compared with the thermostable Tx-Xyn11. The rate of adsorbed enzyme onto lignin was reduced to a similar extent for the P1 and P2 mutants, whereas the adsorption of the P3 mutant was less affected compared with that of Tx-Xyn11. When considered separately, the hydrophobic residues did not affect xylanase adsorption onto lignin. The addition of Tween 20 also led to the decreased adsorption of Tx-Xyn11 onto lignin. These results suggest that hydrophobic interactions are a key parameter in the interaction of Tx-Xyn11 with isolated lignin.

  16. Antioxidant capacity of arabinoxylan oligosaccharide fractions prepared from wheat aleurone using Trichoderma viride or Neocallimastix patriciarum xylanase.

    Science.gov (United States)

    Malunga, Lovemore Nkhata; Beta, Trust

    2015-01-15

    The effect of xylanase type (Trichoderma viride or Neocallimastix patriciarum) and graded ethanol fractionation on the antioxidant capacity (AOC) of arabinoxylan oligosaccharides (AXOS) obtained from wheat aleurone was investigated. AXOS yields were higher using N. patriciarum (62%) than T. viride (44%). The fraction (F100) collected at >80% ethanol concentration constituted 60% of total recovered AXOS. Degree of substitution ranged from 0.20 to 0.60 for ethanol graded fractions. Ferulic acid (FA) esterified to AXOS (8.0 μg/ mg) was 2-fold lower for the N. patriciarum treatment. The mean AOC (41.6, 183.1, and 394.9 μM TE/mg) of T. viride treated AXOS was >1.4-fold higher than N. patriciarum treatment using DPPH and ABTS and ORAC assays, respectively. Fraction F100 had highest AOC. AOC was influenced by the content of esterified FA (R(2)=0.94). The type of xylanase had a major influence on the AOC of the resultant AXOS rich in FA content.

  17. Effect of xylanases on ileal viscosity, intestinal fiber modification, and apparent ileal fiber and nutrient digestibility of rye and wheat in growing pigs.

    Science.gov (United States)

    Lærke, H N; Arent, S; Dalsgaard, S; Bach Knudsen, K E

    2015-09-01

    Two experiments were performed to study the effect of xylanase on ileal extract viscosity, in vivo fiber solubilization and degradation, and apparent ileal digestibility (AID) of fiber constituents, OM, CP, starch, and crude fat in rye and wheat in ileal-cannulated pigs. In Exp. 1, coarse rye without (NX) or with addition of xylanase from Aspergillus niger (AN), (BS), or (TR) was fed to 8 ileal-cannulated barrows (initial BW 30.9 ± 0.3 kg) for 1 wk each according to a double 4 × 4 Latin square design. In Exp. 2, fine rye, fine wheat, and coarse wheat with or without a combination of xylanase from and were fed to 6 ileal-cannulated barrows (initial BW 33.6 ± 0.5 kg) for 1 wk according to a 6 × 6 Latin square design with a 2 × 3 factorial arrangement of enzyme and cereal matrix. Chromic oxide (0.2%) was used as an inert marker. Ileal effluent was collected for 8 h on d 5 and 7 and pooled for analysis. In Exp. 1, TR reduced intestinal viscosity of pigs fed rye from 9.3 mPa·s in the control diet (NX) to 6.0 mPa·s ( fat digestibility of fine wheat with enzyme addition ( < 0.012) in Exp. 2. Collectively, the results suggest that xylanase is more efficient in degrading arabinoxylan from wheat than from rye.

  18. Feed intake, growth, digestibility of dry matter and nitrogen in young pigs as affected by dietary cation-anion difference and supplementation of xylanase

    NARCIS (Netherlands)

    Dersjant-Li, Y.; Schulze, H.; Schrama, J.W.; Verreth, J.A.J.; Verstegen, M.W.A.

    2001-01-01

    An experiment was conducted to test the effect of dietary cation-anion difference (CAD, Na K -Cl, mEq/kg diet) and xylanase addition on feed consumption, digestibility of nutrients, plasma electrolyte balance and growth performance in young pigs. A 2 3 factorial arrangement with three dietary CAD le

  19. Enhanced sugar production from pretreated barley straw by additive xylanase and surfactants in enzymatic hydrolysis for acetone-butanol-ethanol fermentation.

    Science.gov (United States)

    Yang, Ming; Zhang, Junhua; Kuittinen, Suvi; Vepsäläinen, Jouko; Soininen, Pasi; Keinänen, Markku; Pappinen, Ari

    2015-01-01

    This study aims to improve enzymatic sugar production from dilute sulfuric acid-pretreated barley straw for acetone-butanol-ethanol (ABE) fermentation. The effects of additive xylanase and surfactants (polyethylene glycol [PEG] and Tween) in an enzymatic reaction system on straw hydrolysis yields were investigated. By combined application of 2g/100g dry-matter (DM) xylanase and PEG 4000, the glucose yield was increased from 53.2% to 86.9% and the xylose yield was increased from 36.2% to 70.2%, which were considerably higher than results obtained with xylanase or surfactant alone. The ABE fermentation of enzymatic hydrolysate produced 10.8 g/L ABE, in which 7.9 g/L was butanol. The enhanced sugar production increased the ABE yield from 93.8 to 135.0 g/kg pretreated straw. The combined application of xylanase and surfactants has a large potential to improve sugar production from barley straw pretreated with a mild acid and that the hydrolysate showed good fermentability in ABE production.

  20. Effects of diet acidification and xylanase supplementation on performance, nutrient digestibility, duodenal histology and gut microflora of broilers fed wheat based diet

    NARCIS (Netherlands)

    Esmaeilipour, O.; Moravej, H.; Shivazad, M.; Rezaian, M.; Aminzadeh, S.; Krimpen, van M.M.

    2012-01-01

    1. The objective of this experiment was to study the influences of xylanase and citric acid on the performance, nutrient digestibility, digesta viscosity, duodenal histology, and gut microflora of broilers fed on a wheat based diet. 2. The experiment was carried out as a 2 x 3 factorial arrangement

  1. A new chitinase-like xylanase inhibitor protein (XIP from coffee (Coffea arabica affects Soybean Asian rust (Phakopsora pachyrhizi spore germination

    Directory of Open Access Journals (Sweden)

    Mehta Angela

    2011-02-01

    Full Text Available Abstract Background Asian rust (Phakopsora pachyrhizi is a common disease in Brazilian soybean fields and it is difficult to control. To identify a biochemical candidate with potential to combat this disease, a new chitinase-like xylanase inhibitor protein (XIP from coffee (Coffea arabica (CaclXIP leaves was cloned into the pGAPZα-B vector for expression in Pichia pastoris. Results A cDNA encoding a chitinase-like xylanase inhibitor protein (XIP from coffee (Coffea arabica (CaclXIP, was isolated from leaves. The amino acid sequence predicts a (β/α8 topology common to Class III Chitinases (glycoside hydrolase family 18 proteins; GH18, and shares similarity with other GH18 members, although it lacks the glutamic acid residue essential for catalysis, which is replaced by glutamine. CaclXIP was expressed as a recombinant protein in Pichia pastoris. Enzymatic assay showed that purified recombinant CaclXIP had only residual chitinolytic activity. However, it inhibited xylanases from Acrophialophora nainiana by approx. 60% when present at 12:1 (w/w enzyme:inhibitor ratio. Additionally, CaclXIP at 1.5 μg/μL inhibited the germination of spores of Phakopsora pachyrhizi by 45%. Conclusions Our data suggests that CaclXIP belongs to a class of naturally inactive chitinases that have evolved to act in plant cell defence as xylanase inhibitors. Its role on inhibiting germination of fungal spores makes it an eligible candidate gene for the control of Asian rust.

  2. Effects of xylanase and citric acid on the performance, nutrient retention, and characteristics of gastrointestinal tract of broilers fed low-phosphorus wheat-based diets

    NARCIS (Netherlands)

    Esmaeilipour, O.; Shivazad, M.; Moravej, H.; Aminzadeh, S.; Rezaian, M.; Krimpen, van M.M.

    2011-01-01

    An experiment was conducted to study the effects of xylanase and citric acid on the performance, nutrient retention, jejunal viscosity, and size and pH of the gastrointestinal tract of broilers fed a low-P wheat-based diet. The experiment was conducted as a 2 × 3 factorial arrangement with 2 levels

  3. Improvement of the optimum pH of Aspergillus niger xylanase towards an alkaline pH by site-directed mutagenesis.

    Science.gov (United States)

    Li, Fei; Xie, Jingcong; Zhang, Xuesong; Zhao, Linguo

    2015-01-01

    In an attempt to shift the optimal pH of the xylanase B (XynB) from Aspergillus niger towards alkalinity, target mutation sites were selected by alignment between Aspergillus niger xylanase B and other xylanases that have alkalophilic pH optima that highlight charged residues in the eight-residues-longer loop in the alkalophilic xylanase. Multiple engineered XynB mutants were created by site-directed mutagenesis with substitutions Q164K and Q164K+D117N. The variant XynB-117 had the highest optimum pH (at 5.5), which corresponded to a basic 0.5 pH unit shift when compared with the wild-type enzyme. However, the optimal pH of the XynB- 164 mutation was not changed, similar to the wild type. These results suggest that the residues at positions 164 and 117 in the eight-residues-longer loop and the cleft's edge are important in determining the pH optima of XynB from Aspergillus niger.

  4. Expression of the C-terminal family 22 carbohydrate-binding module of xylanase 10B of Clostridium themocellum in tobacco plant

    NARCIS (Netherlands)

    Olawole, O.

    2009-01-01

    Carbohydrate-binding modules have been shown to alter plant cell wall structural architecture. Hence, they have the potential application of being used to engineer the plant to produce tailor-made natural fibers in the cell wall. The Clostridium thermocellum xylanase, Xyn10B, contains two CBMs that

  5. Bis(2,3-dibromo-4,5-dihydroxybenzyl) ether, a marine algae derived bromophenol, inhibits the growth of Botrytis cinerea and interacts with DNA molecules.

    Science.gov (United States)

    Liu, Ming; Wang, Genzhu; Xiao, Lin; Xu, Xuanli; Liu, Xiaohui; Xu, Pingxiang; Lin, Xiukun

    2014-06-27

    Bis(2,3-dibromo-4,5-dihydroxybenzyl) ether (BDDE) is a bromophenol isolated from marine algae. Previous reports have shown that BDDE possesses cytotoxic and antibacterial activity. In the present study, we demonstrate that BDDE displays broad-spectrum antifungal activities, especially on Botrytis cinerea. BDDE inhibits the growth of B. cinerea cultured on a solid medium of potato dextrose agar (PDA) as well as on the potato dextrose broth (PDB) medium. Moreover, BDDE decreases the incidence of fruit decay and severity of strawberries infected with B. cinerea. Further studies have revealed that BDDE decreases the germination rate and inhibits the mycelial growth of B. cinerea. The inhibition mechanisms are related to the disruption of the cell membrane integrity in B. cinerea spores and newly formed germ tubes. This study also suggests that BDDE possibly interacts with DNA via intercalation and minor groove binding. The studies provide evidence that BDDE has potential application in the control of gray mold after fruit harvest and the compound could serve as a candidate or lead template for rational drug design and for the development of antifungal agents.

  6. Bis(2,3-dibromo-4,5-dihydroxybenzyl Ether, a Marine Algae Derived Bromophenol, Inhibits the Growth of Botrytis cinerea and Interacts with DNA Molecules

    Directory of Open Access Journals (Sweden)

    Ming Liu

    2014-06-01

    Full Text Available Bis(2,3-dibromo-4,5-dihydroxybenzyl ether (BDDE is a bromophenol isolated from marine algae. Previous reports have shown that BDDE possesses cytotoxic and antibacterial activity. In the present study, we demonstrate that BDDE displays broad-spectrum antifungal activities, especially on Botrytis cinerea. BDDE inhibits the growth of B. cinerea cultured on a solid medium of potato dextrose agar (PDA as well as on the potato dextrose broth (PDB medium. Moreover, BDDE decreases the incidence of fruit decay and severity of strawberries infected with B. cinerea. Further studies have revealed that BDDE decreases the germination rate and inhibits the mycelial growth of B. cinerea. The inhibition mechanisms are related to the disruption of the cell membrane integrity in B. cinerea spores and newly formed germ tubes. This study also suggests that BDDE possibly interacts with DNA via intercalation and minor groove binding. The studies provide evidence that BDDE has potential application in the control of gray mold after fruit harvest and the compound could serve as a candidate or lead template for rational drug design and for the development of antifungal agents.

  7. Nitrogen fertilization of the host plant influences production and pathogenicity of Botrytis cinerea secondary inoculum.

    Science.gov (United States)

    Abro, Manzoor Ali; Lecompte, François; Bryone, Florian; Nicot, Philippe C

    2013-03-01

    The influence of nitrogen (N) nutrition on a plant's susceptibility to Botrytis spp. and other pathogens is well documented. However, little is known of possible effects on sporulation of the pathogen on diseased tissue and on the pathogenicity of resulting secondary inoculum. To address this question, sporulation by two strains of Botrytis cinerea was quantified on tomato plants produced under different N irrigation regimes with inputs of NO(3)- at 0.5 to 45 mmol liter(-1) (mM). Sporulation decreased significantly (P fertilization up to NO(3)- at 15 to 30 mM. The secondary inoculum was collected and used to inoculate pruning wounds on tomato plants produced under a standard fertilization regime. Pathogenicity of the spores was significantly influenced by the nutritional status of their production substrate. Disease severity was highest with spores produced on plants with very low or very high N fertilization (NO(3)- at 0.5 or 30 mM). It was lowest for inoculum from plants with moderate levels of N fertilization. These results suggest that it may be possible to find an optimum level of N fertilization to reduce the production of secondary inoculum and its pathogenicity to tomato.

  8. Calmodulin Gene Expression in Response to Mechanical Wounding and Botrytis cinerea Infection in Tomato Fruit.

    Science.gov (United States)

    Peng, Hui; Yang, Tianbao; Ii, Wayne M Jurick

    2014-08-29

    Calmodulin, a ubiquitous calcium sensor, plays an important role in decoding stress-triggered intracellular calcium changes and regulates the functions of numerous target proteins involved in various plant physiological responses. To determine the functions of calmodulin in fleshy fruit, expression studies were performed on a family of six calmodulin genes (SlCaMs) in mature-green stage tomato fruit in response to mechanical injury and Botrytis cinerea infection. Both wounding and pathogen inoculation triggered expression of all those genes, with SlCaM2 being the most responsive one to both treatments. Furthermore, all calmodulin genes were upregulated by salicylic acid and methyl jasmonate, two signaling molecules involved in plant immunity. In addition to SlCaM2, SlCaM1 was highly responsive to salicylic acid and methyl jasmonate. However, SlCaM2 exhibited a more rapid and stronger response than SlCaM1. Overexpression of SlCaM2 in tomato fruit enhanced resistance to Botrytis-induced decay, whereas reducing its expression resulted in increased lesion development. These results indicate that calmodulin is a positive regulator of plant defense in fruit by activating defense pathways including salicylate- and jasmonate-signaling pathways, and SlCaM2 is the major calmodulin gene responsible for this event.

  9. Jasmonic acid involves in grape fruit ripening and resistant against Botrytis cinerea.

    Science.gov (United States)

    Jia, Haifeng; Zhang, Cheng; Pervaiz, Tariq; Zhao, Pengcheng; Liu, Zhongjie; Wang, Baoju; Wang, Chen; Zhang, Lin; Fang, Jinggui; Qian, Jianpu

    2016-01-01

    Fruit ripening is a complex process that is regulated by a signal network. Whereas the regulatory mechanism of abscisic acid has been studied extensively in non-climacteric fruit, little is know about other signaling pathways involved in this process. In this study, we performed that plant hormone jasmonic acid plays an important role in grape fruit coloring and softening by increasing the transcription levels of several ripening-related genes, such as the color-related genes PAL1, DFR, CHI, F3H, GST, CHS, and UFGT; softening-related genes PG, PL, PE, Cell, EG1, and XTH1; and aroma-related genes Ecar, QR, and EGS. Lastly, the fruit anthocyanin, phenol, aroma, and cell wall materials were changed. Jasmonic acid positively regulated its biosynthesis pathway genes LOS, AOS, and 12-oxophytodienoate reductase (OPR) and signal pathway genes COI1 and JMT. RNA interference of grape jasmonic acid pathway gene VvAOS in strawberry fruit appeared fruit un-coloring phenotypes; exogenous jasmonic acid rescued this phenotypes. On the contrary, overexpression of grape jasmonic acid receptor VvCOI1 in the strawberry fruit accelerated the fruit-ripening process and induced some plant defense-related gene expression level. Furthermore, jasmonic acid treatment or strong jasmonic acid signal pathway in strawberry fruit make the fruit resistance against Botrytis cinerea.

  10. Induced systemic resistance against Botrytis cinerea by Micromonospora strains isolated from root nodules

    Directory of Open Access Journals (Sweden)

    Pilar eMartínez-Hidalgo

    2015-09-01

    Full Text Available Micromonospora is a Gram positive bacterium that can be isolated from nitrogen fixing nodules from healthy leguminous plants, where they could be beneficial to the plant. Their plant growth promoting activity in legume and non-legume plants has been previously demonstrated. The present study explores the ability of Micromonospora strains to control fungal pathogens and to stimulate plant immunity. Micromonospora strains isolated from surface sterilized nodules of alfalfa showed in vitro antifungal activity against several pathogenic fungi. Moreover, root inoculation of tomato plants with these Micromonospora strains effectively reduced leaf infection by the fungal pathogen Botrytis cinerea, despite spatial separation between both microorganisms. This induced systemic resistance, confirmed in different tomato cultivars, is long lasting. Gene expression analyses evidenced that Micromonospora stimulates the plant capacity to activate defense mechanisms upon pathogen attack. The defensive response of tomato plants inoculated with Micromonospora spp. differs from that of non-inoculated plants, showing a stronger induction of jasmonate-regulated defenses when the plant is challenged with a pathogen. The hypothesis of jasmonates playing a key role in this defense priming effect was confirmed using defense-impaired tomato mutants, since the JA-deficient line def1 was unable to display a long term induced resistance upon Micromonospora spp. inoculation.In conclusion, nodule isolated Micromonospora strains should be considered excellent candidates as biocontrol agents as they combine both direct antifungal activity against plant pathogens and the ability to prime plant immunity.

  11. Lipidperoxidative and proteolytic processes in fungi under the influence of xenobiotica. [Botrytis cinerea; Phytophora infestans

    Energy Technology Data Exchange (ETDEWEB)

    Mueller, H.M.; Edlich, W.; Lyr, H.

    1986-01-01

    As a model system for the investigation of peroxidative effects in the phytopathogenic fungi Botrytis cinerea in submers culture was used. The influence was investigated of hydrogenperoxide with biochemical and cytological methods. The hyphae showed significant changes of the ultrastructure: swelling and vacuolization of mitochondria, loss of ribosomes, vesiculation of endoplasmic reticulum, degradation and lysis of the cell ribosomes, and deposition of lipid droplets on the membranes and in the cytoplasm, dependent on the concentration of the peroxide. Same effects with higher injuries of the membrane system are found in Phytophthorea infestans. A parallel investigation on fibroblasts showed similar effects: pathological changes of mitochondria, but also pycnosis in the nucleus. Comparative ultrastructure investigations were performed in cultures of Phytophthora infestans with tetrachlorcarbon and chloroneb, a specific fungicide substance with a new mode of action. Scavengers like ..cap alpha..-tocopherol and piperonyl butoxide inhibit the toxic effect of the investigated substances in various degrees. Results are discussed in regard to the effect of a lipid peroxidative and proteolytic attack in pesticides of unknown mechanism of action.

  12. Rapid isolation of mycoviral double-stranded RNA from Botrytis cinerea and Saccharomyces cerevisiae

    Directory of Open Access Journals (Sweden)

    Sepúlveda Felipe

    2011-01-01

    Full Text Available Abstract Background In most of the infected fungi, the mycoviruses are latent or cryptic, the infected fungus does not show disease symptoms, and it is phenotypically identical to a non-infected strain of the same species. Because of these properties, the initial stage in the search for fungi infected with mycoviruses is the detection of their viral genome, which in most of the described cases corresponds to double-stranded RNA (dsRNA. So to analyze a large number of fungal isolates it is necessary to have a simple and rapid method to detect dsRNA. Results A rapid method to isolate dsRNA from a virus-infected filamentous fungus, Botrytis cinerea, and from a killer strain of Saccharomyces cerevisiae using commercial minicolumns packed with CF11 cellulose was developed. In addition to being a rapid method, it allows to use small quantities of yeasts or mycelium as starting material, being obtained sufficient dsRNA quantity that can later be analyzed by agarose gel electrophoresis, treated with enzymes for its partial characterization, amplified by RT-PCR and cloned in appropriate vectors for further sequencing. Conclusions The method yields high quality dsRNA, free from DNA and ssRNA. The use of nucleases to degrade the DNA or the ssRNA is not required, and it can be used to isolate dsRNA from any type of fungi or any biological sample that contains dsRNA.

  13. Subsocial Cockroaches Nauphoeta cinerea Mate Indiscriminately with Kin Despite High Costs of Inbreeding

    Science.gov (United States)

    Bouchebti, Sofia; Durier, Virginie; Pasquaretta, Cristian; Rivault, Colette; Lihoreau, Mathieu

    2016-01-01

    Many animals have evolved strategies to reduce risks of inbreeding and its deleterious effects on the progeny. In social arthropods, such as the eusocial ants and bees, inbreeding avoidance is typically achieved by the dispersal of breeders from their native colony. However studies in presocial insects suggest that kin discrimination during mate choice may be a more common mechanism in socially simpler species with no reproductive division of labour. Here we examined this possibility in the subsocial cockroach Nauphoeta cinerea, a model species for research in sexual selection, where males establish dominance hierarchies to access females and control breeding territories. When given a binary choice between a sibling male and a non-sibling male that had the opportunity to establish a hierarchy prior to the tests, females mated preferentially with the dominant male, irrespective of kinship or body size. Despite the lack of kin discrimination during mate choice, inbred-mated females incurred significant fitness costs, producing 20% less offspring than outbred-mated females. We discuss how the social mating system of this territorial cockroach may naturally limit the probability of siblings to encounter and reproduce, without the need for evolving active inbreeding avoidance mechanisms, such as kin recognition. PMID:27655156

  14. Role of outer-membrane proteins and lipopolysaccharide in conjugation between Neisseria gonorrhoeae and Neisseria cinerea.

    Science.gov (United States)

    Genco, C A; Clark, V L

    1988-12-01

    Little is known concerning the mechanism involved in cell contact between the donor and recipient during conjugation in Neisseria gonorrhoeae. The formation of stable mating pairs during conjugation in Escherichia coli appears to require a specific protein as well as LPS in the outer membrane of the recipient cell. To attempt to identify the cell surface components necessary for conjugation in the neisseriae, we began a comparison of the outer membrane of Neisseria cinerea strains that can (Con+) and cannot (Con-) serve as recipients in conjugation with N. gonorrhoeae. There were no differences in outer-membrane protein profiles on SDS-polyacrylamide gel electrophoresis between Con+ and Con- strains that could be correlated with the ability to conjugate. However, whole outer membrane isolated from Con+ strains specifically inhibited conjugation while those from Con- strains did not. Proteolytic cleavage of outer-membrane proteins by trypsin, pronase or alpha-chymotrypsin abolished the inhibitory effect of Con+ outer membranes, suggesting that these outer membranes contained a protease-sensitive protein(s) involved in conjugation. Although periodate oxidation of Con+ outer-membrane carbohydrates did not abolish the inhibitory action of these membranes, purified LPS from both Con+ and Con- strains inhibited conjugation when added at low concentrations. These results suggest that conjugation requires the presence of a specific conjugal receptor that consists of both LPS and one or more outer-membrane proteins. Both Con+ and Con- strains contain the necessary LPS, but only Con+ strains contain the required protein(s).

  15. Induced systemic resistance against Botrytis cinerea by Micromonospora strains isolated from root nodules.

    Science.gov (United States)

    Martínez-Hidalgo, Pilar; García, Juan M; Pozo, María J

    2015-01-01

    Micromonospora is a Gram positive bacterium that can be isolated from nitrogen fixing nodules from healthy leguminous plants, where they could be beneficial to the plant. Their plant growth promoting activity in legume and non-legume plants has been previously demonstrated. The present study explores the ability of Micromonospora strains to control fungal pathogens and to stimulate plant immunity. Micromonospora strains isolated from surface sterilized nodules of alfalfa showed in vitro antifungal activity against several pathogenic fungi. Moreover, root inoculation of tomato plants with these Micromonospora strains effectively reduced leaf infection by the fungal pathogen Botrytis cinerea, despite spatial separation between both microorganisms. This induced systemic resistance, confirmed in different tomato cultivars, is long lasting. Gene expression analyses evidenced that Micromonospora stimulates the plant capacity to activate defense mechanisms upon pathogen attack. The defensive response of tomato plants inoculated with Micromonospora spp. differs from that of non-inoculated plants, showing a stronger induction of jasmonate-regulated defenses when the plant is challenged with a pathogen. The hypothesis of jasmonates playing a key role in this defense priming effect was confirmed using defense-impaired tomato mutants, since the JA-deficient line def1 was unable to display a long term induced resistance upon Micromonospora spp. inoculation. In conclusion, nodule isolated Micromonospora strains should be considered excellent candidates as biocontrol agents as they combine both direct antifungal activity against plant pathogens and the ability to prime plant immunity.

  16. Calmodulin Gene Expression in Response to Mechanical Wounding and Botrytis cinerea Infection in Tomato Fruit

    Directory of Open Access Journals (Sweden)

    Hui Peng

    2014-08-01

    Full Text Available Calmodulin, a ubiquitous calcium sensor, plays an important role in decoding stress-triggered intracellular calcium changes and regulates the functions of numerous target proteins involved in various plant physiological responses. To determine the functions of calmodulin in fleshy fruit, expression studies were performed on a family of six calmodulin genes (SlCaMs in mature-green stage tomato fruit in response to mechanical injury and Botrytis cinerea infection. Both wounding and pathogen inoculation triggered expression of all those genes, with SlCaM2 being the most responsive one to both treatments. Furthermore, all calmodulin genes were upregulated by salicylic acid and methyl jasmonate, two signaling molecules involved in plant immunity. In addition to SlCaM2, SlCaM1 was highly responsive to salicylic acid and methyl jasmonate. However, SlCaM2 exhibited a more rapid and stronger response than SlCaM1. Overexpression of SlCaM2 in tomato fruit enhanced resistance to Botrytis-induced decay, whereas reducing its expression resulted in increased lesion development. These results indicate that calmodulin is a positive regulator of plant defense in fruit by activating defense pathways including salicylate- and jasmonate-signaling pathways, and SlCaM2 is the major calmodulin gene responsible for this event.

  17. Biochemical and Thermodynamic Characterization of a Novel, Low Molecular Weight Xylanase from Bacillus Methylotrophicus CSB40 Isolated from Traditional Korean Food.

    Science.gov (United States)

    Panthi, Sandesh; Choi, Yoon Seok; Choi, Yun Hee; Kim, MiRi; Yoo, Jin Cheol

    2016-04-01

    A low molecular weight xylanase from Bacillus strain CSB40, isolated from traditional Korean food and produced in beechwood xylan, was biochemically and thermodynamically characterized. It was purified 8.12-fold with a 15.88 % yield using DEAE sepharose fast flow, and it was determined to have a mass of ∼27 kDa via sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and xylan zymography. The purified xylanase was optimally active at 50 °C and pH 6 and stable over a wide range of pH (4.5-12.5). The N-terminal amino acid sequence of xylanase was GIQQGDDGKL. The activation energy for beechwood xylan hydrolysis was 29.39 kJmol(-1) with k cat value of 927.582 × 10(2) s(-1). K m and V max were 0.080 mg/ml and 794.63 mmol min(-1) mg(-1). The analysis of other thermodynamic parameters like ∆H, ∆G, ∆S, Q10, ∆GE-S, and ∆GE-T also supported the spontaneous formation of products, greater hydrolytic efficiency, and feasibility of enzymatic reaction, which also ratifies the novelty of this xylanase. The enzyme was strongly activated by Zn(2+) and inhibited by Cu(2+). The principal hydrolyzed end-products of this xylanase are xylobiose, xylotriose, and xylotetrose, which can be used in the pharmaceutical industry and as prebiotic in food.

  18. Cloning of a novel xylanase gene from a newly isolated Fusarium sp. Q7-31 and its expression in Escherichia coli

    Directory of Open Access Journals (Sweden)

    Zhan-Ling Xie

    2012-03-01

    Full Text Available A strain of Q7-31 was isolated from Qinghai-Tibet Plateau and was identified as Fusarium sp. based on its morphological characteristics and ITS rDNA gene sequence analysis. It has the highest capacity of degrading cell wall activity compared with other 11 strains. To do research on its xylanase activity of Fusarium sp. Q7-31 while the degrading the rice cell walls, the complete gene xyn8 that encodes endo-1, 4-β-xylanase secreted by Fusarium sp. Q7-31 was cloned and sequenced. The coding region of the gene is separated by two introns of 56bp and 55bp. It encodes 230 amino acid residues of a protein with a calculated molecular weight of 25.7 kDa. The animo acids sequence of xyn8 gene has higher similarity with those of family 11 of glycosyl hydrolases reported from other microorganisms. The nature peptide encodeing cDNA was subcloned into pGEX5x-1 expression vector. The recombinant plasmid was expressed in Escherichia coli BL21-CodonPlus (DE3-RIL, and xylanase activity was measured. The expression fusion protein was identified by SDS-PAGE and Western blotting, a new specific band of about 52kDa was identified when induced by IPTG. Enzyme activity assay verified the recombinants proteins as a xylanase. A maxium activity of 2.34U/ mg, the xylanase had optimal activity at pH 6.0 and temperature 40ºC .

  19. Control of Botrytis cinerea in Eucalyptus globulus Mini-Cuttings Using Clonostachys and Trichoderma Strains Control de Botrytis cinerea en miniestacas de Eucalyptus globulus Utilizando Cepas de Clonostachys y Trichoderma

    Directory of Open Access Journals (Sweden)

    Salomé Zaldúa

    2010-12-01

    Full Text Available Botrytis cinerea Pers. ex Fr. causes the disease known as gray mold in more than 200 hosts. It is one of the most important pathogens in Chilean forest nurseries and Eucalyptus globulus Labill. is one of the most susceptible species, especially in vegetative reproduction systems. Clonostachys and Trichoderma strains were selected as potential biocontrol agents of gray mold in previous research by the authors. The objective of this study was to evaluate the effectiveness of antagonistic fungi to control B. cinerea in E. globulus mini-cuttings. Five fungi strains were tested and applied weekly, two Clonostachys and three Trichoderma (5 x 10(6 conidia mL-1. In addition, comparison treatments were also used: absolute control (water and fungicide application. The experiment was carried out under operational conditions to produce E. globulus mini-cuttings. The Clonostachys UDC-A10 and UDC-A11 strains reduce mini-cutting mortality caused by B. cinerea in 54 and 71%, respectively, and with effects similar to those achieved by fungicides. Clonostachys UDC-A11 reduces the disease progression rate with the same statistical results as fungicides. A negative effect of applying fungicides on rooting of the surviving mini-cuttings was also confirmed. These results demonstrate the effectiveness of Clonostachys as a control agent against gray mold disease in E. globulus mini-cuttings.Botrytis cinerea Pers. ex Fr. ocasiona la enfermedad conocida como moho gris en más de 200 hospederos. En Chile es uno de los patógenos más importantes en viveros forestales, siendo Eucalyptus globulus Labill. una de las especies más susceptibles, especialmente en los sistemas de reproducción vegetativa. En investigaciones previas, realizadas por los autores, se seleccionaron cepas de Clonostachys y Trichoderma como potenciales agentes de biocontrol del moho gris. El objetivo fue evaluar la eficacia de hongos antagonistas en el control de B. cinerea en mini-estacas de E

  20. Bibliometric Analysis of Literatures on Botrytis cinerea%我国番茄灰霉病文献计量分析

    Institute of Scientific and Technical Information of China (English)

    王欣莹; 宋静; 张冬冬

    2011-01-01

    Based on the data of Botrytis cinerea in China collected by China Journal Full-text Database, the number, year,unit of output, authors and subjects distribution of the literature about Botrytis cinerea were analyzed statistically by the method of literature metrology. Research status and developing trend of Botrytis cinerea in China over the past 20 years were pointed out.%以为数据源,采用文献计量学方法对番茄灰霉病研究文献的数量、文献年份、产出单位、作者及文献研究主题分布等几个方面进行了统计分析,揭示了我国近20年来对番茄灰霉病研究的现状及发展趋势.

  1. Fungistatic activity of Zanthoxylum rhoifolium Lam. bark extracts against fungal plant pathogens and investigation on mechanism of action in Botrytis cinerea.

    Science.gov (United States)

    Carotenuto, Gennaro; Carrieri, Raffaele; Tarantino, Paola; Alfieri, Mariaevelina; Leone, Antonella; De Tommasi, Nunziatina; Lahoz, Ernesto

    2015-01-01

    Plant-derived compounds are emerging as an alternative choice to synthetic fungicides. Chloroform-methanol extract, obtained from the bark of Zanthoxylum rhoifolium, a member of Rutaceae, showed a fungistatic effect on Botrytis cinerea, Sclerotinia sclerotiorum, Alternaria alternata, Colletotrichum gloeosporioides and Clonostachys rosea, when added to the growth medium at different concentrations. A fraction obtained by gel separation and containing the alkaloid O-Methylcapaurine showed significant fungistatic effect against B. cinerea and S. sclerotiorum, two of the most destructive phytopathogenic fungi. The underlying mechanism of such an inhibition was further investigated in B. cinerea, a fungus highly prone to develop fungicide resistance, by analysing the expression levels of a set of genes (BcatrB, P450, CYP51 and TOR). O-Methylcapaurine inhibited the expression of all the analysed genes. In particular, the expression of BcatrB gene, encoding a membrane drug transporter involved in the resistance to a wide range of xenobiotic compounds, was strongly inhibited (91%).

  2. Isolation and Selection of Epiphytic Yeast for Biocontrol of Botrytis cinerea Pers. on Table Grapes Aislación y Selección de Levaduras Epífitas para el Biocontrol de Botrytis cinerea Pers. en Uva de Mesa

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    Marisol Vargas

    2012-09-01

    Full Text Available Botrytis cinerea Pers., the causal agent of gray mold, infects more than 200 plant species. This pathogen has traditionally been controlled by fungicides. However, with the increasing demand for pesticide-free foods new control strategies are needed. The objective of this study was to isolate and select grapevine (Vitis vinifera L. epiphytic yeasts for the biocontrol of B. cinerea in table grapes. Of the total isolated yeasts (n = 256, 32 exhibited mycelial growth inhibition in dual cultures with a halo > 4 mm, and eight of these isolates inhibited > 90% of conidial germination. When evaluating increasing concentrations on conidial germination inhibition, a dose-dependent response was observed with EC90 values from 0.45 x 10(5 to 0.22 x 10(8 cells mL-1. The antagonistic activity of six yeasts against B. cinerea in table grape berries 'Flame Seedless' increased as the yeast colonization time increased from 1 to 24 h on the berries, resulting in a higher biocontrol activity on B. cinerea. These results show the effectiveness of grapevine epiphytic yeasts as biocontrol agents of B. cinerea on table grapes.Botrytis cinerea Pers., agente causal de la pudrición gris, infecta a más de 200 especies vegetales. Tradicionalmente, este patógeno ha sido controlado con fungicidas; sin embargo, la creciente demanda de alimentos libres de pesticidas hace necesario el uso de nuevas estrategias de control. El objetivo de este estudio fue aislar y seleccionar levaduras epífitas de vid (Vitis vinifera L. para el biocontrol de B. cinerea en uva de mesa. Del total de levaduras aisladas (n = 256, 32 presentaron inhibición del crecimiento micelial, en cultivos duales, con un halo > 4 mm y ocho de estos aislamientos inhibieron la germinación de conidias > 90%. Al evaluar concentraciones crecientes de levaduras sobre la inhibición de la germinación de conidias, se observó una respuesta dosis-dependiente, con valores de CE90 de 0,45 x 10(5 a 0,22 x 10(8 c

  3. Epiphyas postvittana (Lepidoptera: Tortricidae)-Botrytis cinerea (Helotiales: Sclerotiniaceae)-Vitis vinifera (Vitales: Vitaceae) Interaction: The Role of B. cinerea on the Development of E. postvittana in Synthetic Nutritional Media.

    Science.gov (United States)

    Rizvi, S Z M; Raman, A

    2015-08-01

    Epiphyas postvittana (Walker) (light-brown apple moth) is a polyphagous herbivore of economic significance, which also feeds on Vitis vinifera L. The E. postvittana-V. vinifera interacting system also involves the participation of the fungus Botrytis cinerea Persoon ex Fries. We have been exploring the relationship among E. postvittana-V. vinifera-B. cinerea over the past two years. In this article, we report the preference and performance of the larvae of E. postvittana raised solely on a synthetic diet incorporated with the mycelial material of B. cinerea (Diet B). To characterize the effect of fungus on the development of E. postvittana, another synthetic diet was prepared that included the lyophilized leaf material of V. vinifera (Diet C). When raised on Diets B and C, a decrease in the duration of larval development and an increase in the survival and fecundity rate of E. postvittana occurred. Diet B influenced the pupal mass, but a significant increase occurred when the larvae were fed on Diet C. The larval emergence rate was the greatest in E. postvittana raised on Diet B, followed by those on Diet C. The F(2) generation of the larvae reared on Diet B showed similar effects as F(1) on the life-history performance of the larvae. Diet B enhanced the life-history performance of E. postvittana, although the larvae of E. postvittana showed little preference to Diet B. The greater fertility rate of E. postvittana reared on Diet B suggests the importance of sterols as shown in Lobesia botrana (Denis & Schiffermüller) (Lepidoptera: Tortricidae) and in a few Myrmicinae (Hymenoptera: Formicidae), which serve as precursors to different ecdysteroids that regulate many critical processes through embryonic development.

  4. 绿菇[ Russula virescens(Schaeff.)Fr.]对灰霉菌(Bortrytis cinerea)的拮抗作用研究%Antagonism Effect of Russula virescens ( Schaeff.) Fr.on Bortrytis cinerea

    Institute of Scientific and Technical Information of China (English)

    黄小琴; 刘勇; 张蕾; 周西全

    2011-01-01

    The antagonism of the fungi Russula virescens(Schaeff. )Fr. To five strains of Bortrytis cinerea was studied. The result showed the antagonism of Russula virescens(Schaeff. ) Fr. To Bortrytis cinerea mainly included hyperparasitic mycelium and the inhibition effect of metabolite. The confrontation ratio ranged from 46.9 % to 52.3 % and mycelium parasitism was visible. The zymotic fluid after cultured Russula virescens(Schaeff. ) Fr from PDB had the strongest effect on Bortrytis cinerea and the growth inhibition rate of the zymotic fluid to Botrytis einerea on pepper was the highest, arrived 61.59 %. Thermal stability of the zymotic fluid was good. Mycelium extract had no effect on Bortrytis einerea.%本文首次研究了绿菇[Russula virescens(schaeff.)Fr.]对5种灰霉菌的拮抗作用.结果表明,绿菇对灰霉菌的拮抗作用主要表现为菌丝体重寄生和代谢产物生长抑制作用,对峙生长抑制率介于46.9%~52.3%,重寄生现象明显;PDB培养基培养绿菇获得的发酵液生长抑制作用最强,对辣椒灰霉菌生长抑制率最高,为61.59%;发酵液热稳定性良好;绿菇菌丝提取液对供试灰霉曹无生长抑制作用.

  5. 灰葡萄孢菌(Botrytis cinerea)基因组中T-DNA整合模式分析%Analysis of T-DNA integration pattern in Botrytis cinerea genome

    Institute of Scientific and Technical Information of China (English)

    刘佳; 张剑云; 朱廷恒; 汪琨; 崔志峰

    2011-01-01

    [Objective]To analyze the T-DNA integration pattern in the genome of grey mold Botrytis cinerea.[Methods]T-DNA ( Transfer DNA) inserted mutant library of Botrytis cinerea was created by Agrobactirium tumfacience mediated transformation.By using TAIL-PCR (Thermal asymmetric interlaced polymerase chain reaction) ,we amplified and cloned the chromosomal regions flanking T-DNA insertions.The obtained T-DNA flanking sequences were subjected to alignment with standard T-DNA border sequence for identification and analysis of integration.[Results]Up to 69% T-DNA inserted at noncoding regions and 30% inserted at exons.Recombination including deletion or addition of bases in T-DNA region was observed.The right borders of the T-DNA were frequently truncated, and by contrast the left borders were less prone to degradation and appeared to have been inserted in a relatively integrated manner.Extra sequence additions also occurred in T-DNA integration sites.[Conclusion]Analysis of T-DNA integration pattern in B.cinerea genome will stimulate the functional genomics study of this fungus.%[目的]研究灰葡萄孢菌(Botrytis cinerea)基因组中T-DNA插入位点的整合模式特征.[方法]利用农杆菌(Agrobactirium tumfacience)介导法构建灰葡萄孢菌T-DNA插入突变体库.利用热不对称交错PCR(TAIL-PCR)技术对转化子中T-DNA的旁侧序列进行扩增和克隆,对获得的旁侧序列进行比对分析.[结果]T-DNA插入在灰葡萄孢菌基因组非编码区的占69%,插入在外显子的占30%.T-DNA在插入的过程中发生了碱基缺失、增加等重组现象,其中左边界(left border,LB)整合到基因组碱基缺失较少,有的保持完整,而右边界(right border,RB)及其近邻的T-DNA区域缺失碱基较多.T-DNA的插入位点还发现有额外的序列插入.[结论]对灰葡萄孢菌中插入T-DNA的整合模式的分析为开展该菌的功能基因组学奠定了基础.

  6. In vitro and in vivo antifungal activities of the essential oils of various plants against tomato grey mould disease agent Botrytis cinerea.

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    Soylu, Emine Mine; Kurt, Sener; Soylu, Soner

    2010-10-15

    The aim of this study was to find an alternative to synthetic fungicides currently used in the control of devastating fungal pathogen Botrytis cinerea, the causal agent of grey mould disease of tomato. Antifungal activities of essential oils obtained from aerial parts of aromatic plants, which belong to the Lamiacea family such as origanum (Origanum syriacum L. var. bevanii), lavender (Lavandula stoechas L. var. stoechas) and rosemary (Rosmarinus officinalis L.), were investigated against B. cinerea. Contact and volatile phase effects of different concentrations of the essential oils were found to inhibit the growth of B. cinerea in a dose-dependent manner. Volatile phase effects of essential oils were consistently found to be more effective on fungal growth than contact phase effect. A volatile vapour of origanum oil at 0.2 μg/ml air was found to completely inhibit the growth of B. cinerea. Complete growth inhibition of pathogen by essential oil of lavender and rosemary was, however, observed at 1.6 μg/ml air concentrations. For the determination of the contact phase effects of the tested essential oils, origanum oil at 12.8 μg/ml was found to inhibit the growth of B. cinerea completely. Essential oils of rosemary and lavender were inhibitory at relatively higher concentrations (25.6 μg/ml). Spore germination and germ tube elongation were also inhibited by the essential oils tested. Light and scanning electron microscopic (SEM) observations revealed that the essential oils cause considerable morphological degenerations of the fungal hyphae such as cytoplasmic coagulation, vacuolations, hyphal shrivelling and protoplast leakage and loss of conidiation. In vivo assays with the origanum essential oil, being the most efficient essential oil, under greenhouse conditions using susceptible tomato plants resulted in good protection against grey mould severity especially as a curative treatment. This study has demonstrated that the essential oils are potential and

  7. Development of an indirect competitive enzyme-linked immunosorbent assay applied to the Botrytis cinerea quantification in tissues of postharvest fruits

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    Raba Julio

    2011-10-01

    Full Text Available Abstract Background Botrytis cinerea is a phytopathogenic fungus responsible for the disease known as gray mold, which causes substantial losses of fruits at postharvest. This fungus is present often as latent infection and an apparently healthy fruit can deteriorate suddenly due to the development of this infection. For this reason, rapid and sensitive methods are necessary for its detection and quantification. This article describes the development of an indirect competitive enzyme-linked immunosorbent assay (ELISA for quantification of B. cinerea in apple (Red Delicious, table grape (pink Moscatel, and pear (William's tissues. Results The method was based in the competition for the binding site of monoclonal antibodies between B. cinerea antigens present in fruit tissues and B. cinerea purified antigens immobilized by a crosslinking agent onto the surface of the microtiter plates. The method was validated considering parameters such as selectivity, linearity, precision, accuracy and sensibility. The calculated detection limit was 0.97 μg mL-1 B. cinerea antigens. The immobilized antigen was perfectly stable for at least 4 months assuring the reproducibility of the assay. The fungus was detected and quantified in any of the fruits tested when the rot was not visible yet. Results were compared with a DNA quantification method and these studies showed good correlation. Conclusions The developed method allowed detects the presence of B. cinerea in asymptomatic fruits and provides the advantages of low cost, easy operation, and short analysis time determination for its possible application in the phytosanitary programs of the fruit industry worldwide.

  8. 山西省灰霉菌对啶酰菌胺的敏感性测定%Sensitivity of Botrytis Cinerea from Shanxi Province to Boscalid

    Institute of Scientific and Technical Information of China (English)

    余玲; 刘慧平; 韩巨才; 张宝俊

    2012-01-01

    啶酰菌胺是一种较新的用于防治灰霉菌的杀菌剂,国内鲜有灰霉菌对该杀菌剂的抗性报道.为了明确山西省灰霉菌对啶酰菌胺敏感性情况,通过菌落直径法测定了山西省8个地区139株灰霉菌对该杀菌剂的敏感性.结果表明,所采菌株中137株为相对敏感菌株,2株表现为低抗,敏感菌株占98%以上,说明山西省灰霉菌对啶酰菌胺具有较高的敏感性,但啶酰菌胺具有中度抗性风险,且灰霉菌极易产生抗药性,故有必要做好抗性预防工作.%Boscalid is a quite new fungicide which is used to control Botrytis cinerea. The report of the resistance to Boscalid in Botrytis cinerea in China is seldom. In order to clear the sensitivity of Botrytis cinerea to Boscalid, inhibition of mycelia growth was used to determine sensitivity of 139 isolates of Botrytis cinerea from 8 areas in Shanxi province. The result showed that 137 isolates were relatively sensitive strains, and 2 strains showed low resistance. The percentage of relatively sensitive strains was over 98%, suggesting that sensitivity of Botrytis cinerea from Shanxi province to Boscalid was relatively high. However, as B. cinerea is a high-risk pathogen and Boscalid is a middle-risk fungicide, appropriate precautions against resistance development should be taken.

  9. Influence of Fungal Strain, Temperature, and Wetness Duration on Infection of Grapevine Inflorescences and Young Berry Clusters by Botrytis cinerea.

    Science.gov (United States)

    Ciliberti, Nicola; Fermaud, Marc; Languasco, Luca; Rossi, Vittorio

    2015-03-01

    The effect of temperature and wetness duration on infection of Vitis vinifera inflorescences (from "inflorescence clearly visible" to "end of flowering" stages) and young berry clusters (at "fruit swelling" and "berries groat-sized" stages) by Botrytis cinerea was investigated. Artificial inoculations were carried out using conidial suspensions of eight B. cinerea strains belonging to the transposon genotypes transposa and vacuma. Infection incidence was significantly affected by strain but not by transposon genotype (transposon genotype accounted for only 6.5% of the variance). Infection incidence was also affected by the interaction between strain and growth stage of the inflorescence or berry cluster (overall accounting for approximately 57% of the experimental variance). Thus, under our experimental conditions, the ability to cause infection was a strain rather than a transposon genotype attribute. Across all strains, infection incidence was lowest when inflorescences were clearly visible or fully developed, highest at flowering (from beginning to end of flowering), and intermediate at the postflowering fruit stages (fruit swelling and berries groat-sized). One transposa strain, however, was highly virulent on all grapevine growth stages tested. The effects of temperature and wetness duration on infection incidence were similar for all fungal strains and grapevine growth stages; infection incidence was highest at 20°C and lowest at 30°C, and was also low at 5°C. Similar results were obtained for mycelial growth and conidial germination. Based on the pooled data for all strains and grapevine growth stages, an equation was developed that accounted for the combined effects of temperature and wetness duration on relative infection incidence. This equation should be useful for developing decision-making systems concerning B. cinerea control at early grapevine growth stages.

  10. Response of direct or priming defense against Botrytis cinerea to methyl jasmonate treatment at different concentrations in grape berries.

    Science.gov (United States)

    Wang, Kaituo; Liao, Yunxia; Kan, Jianquan; Han, Lin; Zheng, Yonghua

    2015-02-02

    This study was conducted to characterize the forms of disease resistance induced by methyl jasmonate (MeJA) in harvested grape berries and to evaluate the impact of the induced resistance on fruit quality. The results showed that MeJA treatment at concentrations from 10 to 100μmol/L could effectively induce disease resistance against Botrytis cinerea and reduce disease incidence in grape berries. The induced disease resistance was tightly associated with increased H2O2 generation, enhanced expression of the defense-related gene VvNPR1.1 and accumulation of stilbene phytoalexins such as tran-resveratrol and its oligomer (trans-)ε-viniferin. The expression of the defense-related gene and synthesis of phytoalexins in 10μmol/L MeJA-treated grape berries were only significantly enhanced upon inoculating the berries with B. cinerea, whereas the 50 or 100μmol/L of MeJA treatment directly induced these defense responses. Hence, we deduce that the low concentration of MeJA (10μmol/L) triggered a priming defense mechanism, while higher concentrations of MeJA (50 or 100μmol/L) directly activated defense responses, thus enhancing disease resistance in grape berries. Moreover, the primed grape berries maintained higher contents of soluble sugars and higher DPPH radical scavenging activity and reducing power compared with those expressing direct defense responses. These results indicate that priming of defense is a cost-effective strategy to protect harvested grape berries from B. cinerea infection in terms of minimizing quality loss.

  11. The signalling mucin Msb2 regulates surface sensing and host penetration via BMP1 MAP kinase signalling in Botrytis cinerea.

    Science.gov (United States)

    Leroch, Michaela; Mueller, Nathalie; Hinsenkamp, Isabel; Hahn, Matthias

    2015-10-01

    Botrytis cinerea is a necrotrophic fungus that infects a wide range of fruit, vegetable and flower crops. Penetration of the host cuticle occurs via infection structures that are formed in response to appropriate plant surface signals. The differentiation of these structures requires a highly conserved mitogen-activated protein (MAP) kinase cascade including the MAP kinase BMP1. In yeast and several plant-pathogenic fungi, the signalling mucin Msb2 has been shown to be involved in surface recognition and MAP kinase activation. In this study, a B. cinerea msb2 mutant was generated and characterized. The mutant showed normal growth, sporulation, sclerotia formation and stress resistance. In the absence of nutrients, abnormal germination with multiple germ tubes was observed. In the presence of sugars, normal germination occurred, but msb2 germlings were almost unable to form appressoria or infection cushions on hard surfaces. Nevertheless, the msb2 mutant showed only a moderate delay in lesion formation on different host plants, and formed expanding lesions similar to the wild-type. Although the wild-type showed increasing BMP1 phosphorylation during the first hours of germination on hard surfaces, the phosphorylation levels in the msb2 mutant were strongly reduced. Several genes encoding secreted proteins were found to be co-regulated by BMP1 and Msb2 during germination. Taken together, B. cinerea Msb2 is likely to represent a hard surface sensor of germlings and hyphae that triggers infection structure formation via the activation of the BMP1 MAP kinase pathway.

  12. Dark Period Following UV-C Treatment Enhances Killing of Botrytis cinerea Conidia and Controls Gray Mold of Strawberries.

    Science.gov (United States)

    Janisiewicz, Wojciech J; Takeda, Fumiomi; Glenn, D Michael; Camp, Mary J; Jurick, Wayne M

    2016-04-01

    Strawberries are available throughout the year either from production in the field or from high and low tunnel culture. Diversity of production conditions results in new challenges in controlling diseases before and after harvest. Fungicides have traditionally been used to control these diseases; however, their limitations necessitate a search for new approaches. We found that UV-C irradiation of Botrytis cinerea, a major pathogen of strawberry, can effectively kill this fungus if a dark period follows the treatment. The inclusion of a 4-h dark period resulted in almost complete kill of B. cinerea conidia on agar media at a dose of 12.36 J/m2. The UV-C dose did not cause a reduction in photosynthesis in strawberry leaves or discoloration of sepals, even after exposing plants repeatedly (twice a week) for 7 weeks. Although irradiation of dry conidia of B. cinerea with this dose resulted in some survival, the conidia were not infective and not able to cause decay even when inoculated onto a highly susceptible mature apple fruit. Irradiation of strawberry pollen at 12.36 J/m2 did not affect pollen germination, tube growth and length in vitro, or germination and tube growth in the style of hand-pollinated emasculated strawberry flowers. No negative effect of the UV-C treatment was observed on fruit yield and quality in high tunnel culture. In the fruit and flower petal inoculation tests, the UV-C treatment was highly effective in reducing fruit decay and petal infection. This UV-C treatment with an exposure time of 60 s may be useful in controlling gray mold in tunnel production of strawberries and may also have the potential for use in intensive field and indoor production of other fruits and vegetables providing that a 4-h dark period follows the irradiation.

  13. Induction of Volatile Organic Compounds of Lycopersicon esculentum Mill. and Its Resistance to Botrytis cinerea Pers. by Burdock Oligosaccharide

    Institute of Scientific and Technical Information of China (English)

    Pei-Qing He; Li Tian; Kao-Shan Chen; Lin-Hua Hao; Guang-You Li

    2006-01-01

    In the present study, we investigated the induction of volatile organic compounds (VOCs) of Lycopersicon esculentum Mill. and its resistance to Botrytis cinerea Pers. by burdock oligosaccharide. The disease severity of L. esculentum was evaluated 48 h after treatment with 0.6% burdock oligosaccharide, followed by inoculation with a spore suspension of B. cinerea. The formation of O()2, the activity of lipoxygenases (LOX), peroxidase (POD), catalase (CAT), and superoxide dismutase (SOD), and the quantity and quality of changes in VOCs were determined a period of time after treatment with 0.6% burdock oligosaccharide. The results demonstrated that the disease index in treated plants was decreased by 42.5% compared with control 96 h after inoculation. The production of O()2 reached a maximum 6 h after treatment (1.36-fold compared with control). There was an increase in LOX, POD, CAT and SOD activity in response to burdock oligosaccharide treatment and the enzymes showed different trends in the time-course of induction. At 120 h after treatment,(E)-2-hexenal was increased by 92% compared with control, whereas methyl salicylate showed a gradual increase with induction period. Previous results had demonstrated that chitosan elicitor enhanced the production VOCs of L. esculentum and decreased plant susceptibility towards B. cinerea. Together, these findings suggest that increasing the production of VOCs in response to burrdock oligosaccharide may be an important mechanism for L. esculentumin its defense against pathogens. In addition, burrdock oligosaccharide may act as a potent elicitor of resistance to disease in L. esculentum.

  14. Overexpression of Three Glucosinolate Biosynthesis Genes in Brassica napus Identifies Enhanced Resistance to Sclerotinia sclerotiorum and Botrytis cinerea.

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    Yuanyuan Zhang

    Full Text Available Sclerotinia sclerotiorum and Botrytis cinerea are notorious plant pathogenic fungi with an extensive host range including Brassica crops. Glucosinolates (GSLs are an important group of secondary metabolites characteristic of the Brassicales order, whose degradation products are proving to be increasingly important in plant protection. Enhancing the defense effect of GSL and their associated degradation products is an attractive strategy to strengthen the resistance of plants by transgenic approaches. We generated the lines of Brassica napus with three biosynthesis genes involved in GSL metabolic pathway (BnMAM1, BnCYP83A1 and BnUGT74B1, respectively. We then measured the foliar GSLs of each transgenic lines and inoculated them with S. sclerotiorum and B. cinerea. Compared with the wild type control, over-expressing BnUGT74B1 in B. napus increased the aliphatic and indolic GSL levels by 1.7 and 1.5 folds in leaves respectively; while over-expressing BnMAM1 or BnCYP83A1 resulted in an approximate 1.5-fold higher only in the aliphatic GSL level in leaves. The results of plant inoculation demonstrated that BnUGT74B1-overexpressing lines showed less severe disease symptoms and tissue damage compared with the wild type control, but BnMAM1 or BnCYP83A1-overexpressing lines showed no significant difference in comparison to the controls. These results suggest that the resistance to S. sclerotiorum and B. cinerea in B. napus could be enhanced through tailoring the GSL profiles by transgenic approaches or molecular breeding, which provides useful information to assist plant breeders to design improved breeding strategies.

  15. Functional analysis of BcBem1 and its interaction partners in Botrytis cinerea: impact on differentiation and virulence.

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    Sabine Giesbert

    Full Text Available In phytopathogenic fungi the establishment and maintenance of polarity is not only essential for vegetative growth and differentiation, but also for penetration and colonization of host tissues. We investigated orthologs of members of the yeast polarity complex in the grey mould fungus Botrytis cinerea: the scaffold proteins Bem1 and Far1, the GEF (guanine nucleotide exchange factor Cdc24, and the formin Bni1 (named Sep1 in B. cinerea. BcBem1 does not play an important role in regular hyphal growth, but has significant impact on spore formation and germination, on the establishment of conidial anastomosis tubes (CATs and on virulence. As in other fungi, BcBem1 interacts with the GEF BcCdc24 and the formin BcSep1, indicating that in B. cinerea the apical complex has a similar structure as in yeast. A functional analysis of BcCdc24 suggests that it is essential for growth, since it was not possible to obtain homokaryotic deletion mutants. Heterokaryons of Δcdc24 (supposed to exhibit reduced bccdc24 transcript levels already show a strong phenotype: an inability to penetrate the host tissue, a significantly reduced growth rate and malformation of conidia, which tend to burst as observed for Δbcbem1. Also the formin BcSep1 has significant impact on hyphal growth and development, whereas the role of the putative ortholog of the yeast scaffold protein Far1 remains open: Δbcfar1 mutants have no obvious phenotypes.

  16. Molecular characterization of pyraclostrobin resistance and structural diversity of the cytochrome b gene in Botrytis cinerea from apple.

    Science.gov (United States)

    Yin, Y N; Kim, Y K; Xiao, C L

    2012-03-01

    Botrytis cinerea isolates obtained from apple orchards were screened for resistance to the quinone outside inhibitor (QoI) pyraclostrobin. Of the 220 isolates tested, 43 (19.5%) were resistant to pyraclostrobin. Analysis of partial sequences of the cytochrome b gene (cyt b) in five pyraclostrobin-resistant (PR) and five pyraclostrobin-sensitive (PS) isolates showed that PR isolates harbored the point mutation leading to the substitution of glycine by alanine at codon position 143 in cyt b (G143A). Two pairs of allele-specific primers were designed based on this point mutation, and allele-specific polymerase chain reaction analysis with these primers showed that all 73 PR isolates (including 30 collected from decayed apple fruit) harbored the G143A mutation but PS isolates did not. Six pairs of primers were designed to analyze the presence of various introns in cyt b. There were six types (I to VI) of cyt b present in 247 isolates of B. cinerea collected from various apple-production areas in Washington State. Of the 247 isolates, 23 had type I cyt b containing all four introns (Bcbi-67/68, Bcbi-131/132, Bcbi-143/144, and Bcbi-164), 176 had type II cyt b containing three introns (Bcbi-67/68, Bcbi-131/132, and Bcbi-164), six had type III cyt b containing two introns (Bcbi-67/68 and Bcbi-131/132), one had type IV cyt b containing two introns (Bcbi-131/132 and Bcbi-164), one had type V cyt b containing only the Bcbi-131/132 intron, and 40 had type VI cyt b containing no introns. This is the first report of types III to VI cyt b present in B. cinerea. All 73 PR isolates did not carry the Bcbi-143/144 intron in cyt b. Of the 247 isolates tested, >90% did not carry the Bcbi-143/144 intron in cyt b, suggesting that B. cinerea populations from apple pose a high inherent risk for the development of resistance to QoIs because the presence of this intron in cyt b prevents the occurrence of G143A-mediated resistance. Analysis of genetic background based on three microsatellite

  17. Expression in E. coli and characterization of the catalytic domain of Botrytis cinerea chitin synthase

    Directory of Open Access Journals (Sweden)

    Piffeteau Annie

    2010-11-01

    Full Text Available Abstract Background Chitin synthase 3a (CHS3a from Botrytis cinerea (Bc catalyses the multiple transfer of N-acetylglucosamine (GlcNAc residues to the growing chitin chain. Chitin, a β-1,4 linked GlcNAc homopolymer, is an essential cell wall component of filamentous fungi. Chitin synthase, processive membranous protein, has been recognized as a promising target for new antifungicides. Enzymatic characterizations of chitin synthases have been limited, mainly because purity and amounts of integral enzyme obtained after purification procedures have not been sufficient. Findings We undertook the preparation of two BcCHS3a fragment proteins, containing only the central domain and devoid of the N-terminal and transmembrane C-terminal regions. The central domain of CHS3a, named SGC (Spsa GntI Core, is conserved in all UDP-glycosyltransferases and it is believed to contain the active site of the enzyme. CHS3a-SGC protein was totally expressed as inclusion bodies in Escherichia coli. We performed recombinant CHS3a-SGC purification in denaturing conditions, followed by a refolding step. Although circular dichroism spectra clearly exhibited secondary structures of renatured CHS3a-SGC, no chitin synthase activity was detected. Nevertheless CHS3a-SGC proteins show specific binding for the substrate UDP-GlcNAc with a dissociation constant similar to the Michaelis constant and a major contribution of the uracil moiety for recognition was confirmed. Conclusions Milligram-scale quantities of CHS3a-SGC protein with native-like properties such as specific substrate UDP-GlcNAc binding could be easily obtained. These results are encouraging for subsequent heterologous expression of full-length CHS3a.

  18. Transcriptome and metabolome reprogramming in Vitis vinifera cv. Trincadeira berries upon infection with Botrytis cinerea.

    Science.gov (United States)

    Agudelo-Romero, Patricia; Erban, Alexander; Rego, Cecília; Carbonell-Bejerano, Pablo; Nascimento, Teresa; Sousa, Lisete; Martínez-Zapater, José M; Kopka, Joachim; Fortes, Ana Margarida

    2015-04-01

    Vitis vinifera berries are sensitive towards infection by the necrotrophic pathogen Botrytis cinerea, leading to important economic losses worldwide. The combined analysis of the transcriptome and metabolome associated with fungal infection has not been performed previously in grapes or in another fleshy fruit. In an attempt to identify the molecular and metabolic mechanisms associated with the infection, peppercorn-sized fruits were infected in the field. Green and veraison berries were collected following infection for microarray analysis complemented with metabolic profiling of primary and other soluble metabolites and of volatile emissions. The results provided evidence of a reprogramming of carbohydrate and lipid metabolisms towards increased synthesis of secondary metabolites involved in plant defence, such as trans-resveratrol and gallic acid. This response was already activated in infected green berries with the putative involvement of jasmonic acid, ethylene, polyamines, and auxins, whereas salicylic acid did not seem to be involved. Genes encoding WRKY transcription factors, pathogenesis-related proteins, glutathione S-transferase, stilbene synthase, and phenylalanine ammonia-lyase were upregulated in infected berries. However, salicylic acid signalling was activated in healthy ripening berries along with the expression of proteins of the NBS-LRR superfamily and protein kinases, suggesting that the pathogen is able to shut down defences existing in healthy ripening berries. Furthermore, this study provided metabolic biomarkers of infection such as azelaic acid, a substance known to prime plant defence responses, arabitol, ribitol, 4-amino butanoic acid, 1-O-methyl- glucopyranoside, and several fatty acids that alone or in combination can be used to monitor Botrytis infection early in the vineyard.

  19. Chemical Characterization of Different Sumac and Pomegranate Extracts Effective against Botrytis cinerea Rots.

    Science.gov (United States)

    Romeo, Flora V; Ballistreri, Gabriele; Fabroni, Simona; Pangallo, Sonia; Nicosia, Maria Giulia Li Destri; Schena, Leonardo; Rapisarda, Paolo

    2015-06-30

    Pomegranate (Punica granatum L.) peel and sumac (Rhus coriaria L.) fruit and leaf extracts were chemically characterized and their ability to inhibit table grape (cv. Italia) rots caused by Botrytis cinerea was evaluated on artificially inoculated berries. Different extraction methods were applied and extracts were characterized through Ultra Fast High Performance Liquid Chromatography coupled to Photodiode array detector and Electrospray ionization Mass spectrometer (UPLC-PDA-ESI/MSn) for their phenol and anthocyanin contents. The concentrated pomegranate peel extract (PGE-C) was the richest in phenols (66.97 g gallic acid equivalents/kg) while the concentrated sumac extract from fruits (SUF-C) showed the highest anthocyanin amount (171.96 mg cyanidin 3-glucoside equivalents/kg). Both phenolic and anthocyanin profile of pomegranate and sumac extracts were quite different: pomegranate extract was rich in cyanidin 3-glucoside, pelargonidin 3-glucoside and ellagic acid derivatives, while sumac extract was characterized by 7-methyl-cyanidin 3-galactoside and gallic acid derivatives. The concentrated extracts from both pomegranate peel and sumac leaves significantly reduced the development of Botrytis rots. In particular, the extract from pomegranate peel completely inhibited the pathogen at different intervals of time (0, 12, and 24 h) between treatment and pathogen inoculation on fruits maintained at 22-24 °C and high relative humidity (RH). This extract may represent a valuable alternative to control postharvest fungal rots in view of its high efficacy because of the low cost of pomegranate peel, which is a waste product of processing factories.

  20. Fitness and Competitive Ability of Botrytis cinerea Isolates with Resistance to Multiple Chemical Classes of Fungicides.

    Science.gov (United States)

    Chen, S N; Luo, C X; Hu, M J; Schnabel, G

    2016-09-01

    Resistance to multiple chemical classes of fungicides in Botrytis cinerea isolates from eastern United States strawberry fields is common and strategies to control them are needed. In this study, we compared fitness and competitive ability of eight sensitive isolates (S), eight isolates resistant to five or six chemical classes of fungicides but not to phenylpyrroles (5CCR), and eight isolates resistant to six or seven chemical classes including phenylpyrroles (6CCR/MDR1h). The latter included the MDR1h phenotype due to overexpression of atrB based on Δ497V/L in mrr1. The 6CCR/MDR1h isolates grew more slowly at 4°C on potato dextrose agar, and both 5CCR and 6CCR/MDR1h isolates were hypersensitive to osmotic stress compared with S isolates. In contrast, no differences were found in oxidative sensitivity, aggressiveness, and spore production in vivo, and sclerotia production and viability in vitro. In competition experiments, the 5CCR and 6CCR/MDR1h isolates were both outcompeted by S isolates and 6CCR/MDR1h isolates were outcompeted by 5CCR isolates in the absence of fungicide pressure. Under selective pressure of a fludioxonil/pyraclostrobin rotation, the 6CCR/MDR1h isolates dominated after coinoculation with 5CCR and S isolates. The competitive disadvantage of 5CCR and especially 6CCR/MDR1h isolates suggest that, in the absence of fungicide selection pressure, S isolates may reduce inoculum potential of multifungicide-resistant isolates under field conditions.

  1. Chemical Characterization of Different Sumac and Pomegranate Extracts Effective against Botrytis cinerea Rots

    Directory of Open Access Journals (Sweden)

    Flora V. Romeo

    2015-06-01

    Full Text Available Pomegranate (Punica granatum L. peel and sumac (Rhus coriaria L. fruit and leaf extracts were chemically characterized and their ability to inhibit table grape (cv. Italia rots caused by Botrytis cinerea was evaluated on artificially inoculated berries. Different extraction methods were applied and extracts were characterized through Ultra Fast High Performance Liquid Chromatography coupled to Photodiode array detector and Electrospray ionization Mass spectrometer (UPLC-PDA-ESI/MSn for their phenol and anthocyanin contents. The concentrated pomegranate peel extract (PGE-C was the richest in phenols (66.97 g gallic acid equivalents/kg while the concentrated sumac extract from fruits (SUF-C showed the highest anthocyanin amount (171.96 mg cyanidin 3-glucoside equivalents/kg. Both phenolic and anthocyanin profile of pomegranate and sumac extracts were quite different: pomegranate extract was rich in cyanidin 3-glucoside, pelargonidin 3-glucoside and ellagic acid derivatives, while sumac extract was characterized by 7-methyl-cyanidin 3-galactoside and gallic acid derivatives. The concentrated extracts from both pomegranate peel and sumac leaves significantly reduced the development of Botrytis rots. In particular, the extract from pomegranate peel completely inhibited the pathogen at different intervals of time (0, 12, and 24 h between treatment and pathogen inoculation on fruits maintained at 22–24 °C and high relative humidity (RH. This extract may represent a valuable alternative to control postharvest fungal rots in view of its high efficacy because of the low cost of pomegranate peel, which is a waste product of processing factories.

  2. Biochemical and thermodynamic characteristics of thermo-alkali-stable xylanase from a novel polyextremophilic Bacillus halodurans TSEV1.

    Science.gov (United States)

    Kumar, Vikash; Satyanarayana, T

    2013-09-01

    The purified extracellular xylanase of polyextremophilic Bacillus halodurans TSEV1 has been visualized as a single band on SDS-PAGE and eluted as single peak by gel filtration, with a molecular mass of 40 kDa. The peptide finger print and cloned xylanase gene sequence analyses indicate that this enzyme belongs to GH family 10. The active site carboxyl residues are mainly involved in catalysis, while tryptophan residues are involved in substrate binding. The enzyme is optimally active at 80 °C and pH 9.0, and stable in the pH range of 7.0-12.0 with T 1/2 of 35 min at 80 °C (pH 9.0). Activation energy for birch wood xylan hydrolysis is 30.51 kJ mol(-1). The K m, V max and k cat (birchwood xylan) are 2.05 mg ml(-1), 333.33 μmol mg(-1 )min(-1) and 3.33 × 10(4) min(-1), respectively. The pKa1 and pKa2 of ionizable groups of the active site that influence V max are 8.51 and 11.0. The analysis of thermodynamic parameters for xylan hydrolysis suggests this as a spontaneous process. The enzyme is resistant to chemical denaturants like urea and guanidinium-HCl. The site-directed mutagenesis of catalytic glutamic acid residues (E196 and E301) resulted in a complete loss of activity. The birch wood xylan hydrolyzate contained xylobiose and xylotriose as the main products without any trace of xylose, and the enzyme hydrolyzes xylotetraose and xylopentaose rapidly to xylobiose. Thermo-alkali-stability, resistance to various chemical denaturants and mode of action make it a useful biocatalyst for generating xylo-oligosaccharides from agro-residues and bleaching of pulp in paper industries.

  3. Negative regulation of ABA signaling by WRKY33 is critical for Arabidopsis immunity towards Botrytis cinerea 2100.

    Science.gov (United States)

    Liu, Shouan; Kracher, Barbara; Ziegler, Jörg; Birkenbihl, Rainer P; Somssich, Imre E

    2015-06-15

    The Arabidopsis mutant wrky33 is highly susceptible to Botrytis cinerea. We identified >1680 Botrytis-induced WRKY33 binding sites associated with 1576 Arabidopsis genes. Transcriptional profiling defined 318 functional direct target genes at 14 hr post inoculation. Comparative analyses revealed that WRKY33 possesses dual functionality acting either as a repressor or as an activator in a promoter-context dependent manner. We confirmed known WRKY33 targets involved in hormone signaling and phytoalexin biosynthesis, but also uncovered a novel negative role of abscisic acid (ABA) in resistance towards B. cinerea 2100. The ABA biosynthesis genes NCED3 and NCED5 were identified as direct targets required for WRKY33-mediated resistance. Loss-of-WRKY33 function resulted in elevated ABA levels and genetic studies confirmed that WRKY33 acts upstream of NCED3/NCED5 to negatively regulate ABA biosynthesis. This study provides the first detailed view of the genome-wide contribution of a specific plant transcription factor in modulating the transcriptional network associated with plant immunity.

  4. Tomato transcriptome and mutant analyses suggest a role for plant stress hormones in the interaction between fruit and Botrytis cinerea

    Directory of Open Access Journals (Sweden)

    Barbara eBlanco-Ulate

    2013-05-01

    Full Text Available Fruit-pathogen interactions are a valuable biological system to study the role of plant development in the transition from resistance to susceptibility. In general, unripe fruit are resistant to pathogen infection but become increasingly more susceptible as they ripen. During ripening, fruit undergo significant physiological and biochemical changes that are coordinated by complex regulatory and hormonal signaling networks. The interplay between multiple plant stress hormones in the interaction between plant vegetative tissues and microbial pathogens has been documented extensively, but the relevance of these hormones during infections of fruit is unclear. In this work, we analyzed a transcriptome study of tomato fruit infected with Botrytis cinerea in order to profile the expression of genes for the biosynthesis, modification and signal transduction of ethylene (ET, salicylic acid (SA, jasmonic acid (JA, and abscisic acid (ABA, hormones that may be not only involved in ripening, but also in fruit interactions with pathogens. The changes in relative expression of key genes during infection and assays of susceptibility of fruit with impaired synthesis or perception of these hormones were used to formulate hypotheses regarding the involvement of these regulators in the outcome of the tomato fruit-B. cinerea interaction.

  5. Antimicrobial Activity of Emilia sonchifolia DC., Tridax procumbens L. and Vernonia cinerea L. of Asteracea Family: Potential as Food Preservatives.

    Science.gov (United States)

    Yoga Latha, L; Darah, I; Sasidharan, S; Jain, K

    2009-09-01

    Chemical preservatives have been used in the food industry for many years. However, with increased health concerns, consumers prefer additive-free products or food preservatives based on natural products. This study evaluated antimicrobial activities of extracts from Emilia sonchifolia L. (Common name: lilac tassel flower), Tridax procumbens L. (Common name: tridax daisy) and Vernonia cinerea L. (Common name: Sahadevi), belonging to the Asteracea family, to explore their potential for use against general food spoilage and human pathogens so that new food preservatives may be developed. Three methanol extracts of these plants were tested in vitro against 20 bacterial species, 3 yeast species, and 12 filamentous fungi by the agar diffusion and broth dilution methods. The V. cinerea extract was found to be most effective against all of the tested organisms and the methanol fraction showed the most significant (p extracts determined by the broth dilution method ranged from 1.56 to 100.00mg/mL. The MIC of methanol fraction was the lowest in comparison to the other four extracts. The study findings indicate that bioactive natural products from these plants may be isolated for further testing as leads in the development of new pharmaceuticals in food preservation as well as natural plant-based medicine.

  6. Characterization of endophytic Bacillus strains from tomato plants (Lycopersicon esculentum) displaying antifungal activity against Botrytis cinerea Pers.

    Science.gov (United States)

    Kefi, Asma; Ben Slimene, Imen; Karkouch, Ines; Rihouey, Christophe; Azaeiz, Sana; Bejaoui, Marwa; Belaid, Rania; Cosette, Pascal; Jouenne, Thierry; Limam, Ferid

    2015-12-01

    Eighty endophytic bacteria were isolated from healthy tissues of roots, stems, leaves and fruits of tomato plants (Lycopersicon esculentum). Four strains, named BL1, BT5, BR8 and BF11 were selected for their antagonism against Botrytis cinerea, a phytopathogenic fungus responsible of gray mold in several important crops, with growth inhibitory activity ranging from 27 to 53%. Morphological, biochemical, and molecular parameters as 16S rDNA sequencing demonstrated that the selected bacterial strains were related to Bacillus species which are known to produce and secrete a lot of lipopeptides with strong inhibitory effect against pathogen mycelial growth. Electrospray mass spectrometry analysis showed that these strains produced heterogeneous mixture of antibiotics belonging to fengycin and surfactin for BL1 and BT5, to iturin and surfactin for BR8, to bacillomycin D, fengycin and surfactin for BF11. Furthermore, these bacteria exhibited biocontrol potential by reducing the disease severity when tested on detached leaflets. Based on their antifungal activity against Botrytis cinerea, these strains could be used for biological control of plant diseases.

  7. Tomato transcriptome and mutant analyses suggest a role for plant stress hormones in the interaction between fruit and Botrytis cinerea.

    Science.gov (United States)

    Blanco-Ulate, Barbara; Vincenti, Estefania; Powell, Ann L T; Cantu, Dario

    2013-01-01

    Fruit-pathogen interactions are a valuable biological system to study the role of plant development in the transition from resistance to susceptibility. In general, unripe fruit are resistant to pathogen infection but become increasingly more susceptible as they ripen. During ripening, fruit undergo significant physiological and biochemical changes that are coordinated by complex regulatory and hormonal signaling networks. The interplay between multiple plant stress hormones in the interaction between plant vegetative tissues and microbial pathogens has been documented extensively, but the relevance of these hormones during infections of fruit is unclear. In this work, we analyzed a transcriptome study of tomato fruit infected with Botrytis cinerea in order to profile the expression of genes for the biosynthesis, modification and signal transduction of ethylene (ET), salicylic acid (SA), jasmonic acid (JA), and abscisic acid (ABA), hormones that may be not only involved in ripening, but also in fruit interactions with pathogens. The changes in relative expression of key genes during infection and assays of susceptibility of fruit with impaired synthesis or perception of these hormones were used to formulate hypotheses regarding the involvement of these regulators in the outcome of the tomato fruit-B. cinerea interaction.

  8. Synergistic effect of the combined treatment with gamma irradiation and sodium dichloroisocyanurate to control gray mold (Botrytis cinerea) on paprika

    Science.gov (United States)

    Yoon, Minchul; Jung, Koo; Lee, Kwang-Youll; Jeong, Je-Yong; Lee, Ju-Woon; Park, Hae-Jun

    2014-05-01

    Gray mold (Botrytis cinerea) is one of the most major fungal pathogens in paprika. Generally, gamma irradiation over 1 kGy is effective for the control of fungal pathogens; however, a significant change in fruit quality (physical properties) on paprika was shown from gamma irradiation at over 0.6 kGy (pcinerea isolated from naturally-infected postharvest paprika, fungal symptoms were observed in the stem and exocarp of paprika after conidial inoculation. From the sensitivity of gamma irradiation and NaDCC, B. cinerea conidia were fully inactivated by 4 kGy of gamma irradiation (D10 value 0.99 kGy), and were fully inactivated by 50 ppm NaDCC treatment. The fungal symptoms were not detected by the dose-dependent gamma irradiation (>4 kGy) and NaDCC (>50 ppm). As a result of the combined treatment of gamma irradiation and NaDCC, the D10 value was significantly reduced by 1.06, 0.88, 0.77, and 0.58 kGy (pfruits.

  9. Suppression of the homeobox gene HDTF1 enhances resistance to Verticillium dahliae and Botrytis cinerea in cotton

    Institute of Scientific and Technical Information of China (English)

    Wei Gao; Lu Long; Li Xu; Keith Lindsey; Xianlong Zhang; Longfu Zhu

    2016-01-01

    Development of pathogen-resistant crops, such as fungus-resistant cotton, has significantly reduced chemical application and improved crop yield and quality. However, the mechanism of resistance to cotton pathogens such as Verticillium dahliae is still poorly understood. In this study, we characterized a cotton gene (HDTF1) that was isolated following transcriptome profiling during the resistance response of cotton to V. dahliae. HDTF1 putatively encodes a homeodomain transcription factor, and its expression was found to be down-regulated in cotton upon inoculation with V. dahliae and Botrytis cinerea. To characterise the involvement of HDTF1 in the response to these pathogens, we used virus-induced gene silencing (VIGS) to generate HDTF1-silenced cotton. VIGS reduction in HDTF1 expression significantly enhanced cotton plant resistance to both pathogens. HDTF1 silencing resulted in activation of jasmonic acid (JA)-mediated signaling and JA accumulation. However, the silenced plants were not altered in the accumulation of salicylic acid (SA) or the expression of marker genes associated with SA signaling. These results suggest that HDTF1 is a negative regulator of the JA pathway, and resistance to V. dahliae and B. cinerea can be engineered by activation of JA signaling.

  10. Antagonism in vitro of bacterial isolates from comercial and wild strawberry vs. Botrytis cinerea and Rhizopus stolonifer

    Directory of Open Access Journals (Sweden)

    Rosa Isela Plascencia Tenorio

    2012-09-01

    Full Text Available Strawberry is a non-climacteric fruit, with a short postharvest life. The loss of fruit quality may be due, among other factors to damage caused by pathogens. Among the most common fungi are causing gray mold (Botrytis cinerea and white rot (Rhizopus stolonifer two phytopathogenic impact on their growth rate which allows you to colonize the surface of these caused major economic losses. An alternative to control damage in fruit postharvest pathogens usingmicrobial antagonists may be present in the plant or fruit, but at low densities. In this study bacteria were isolated from leaf tissue and wild strawberry fruit (Duchesnea indicates Andr. Fock and comercial strawberry. Those isolates that were selected had the highest percentages of inhibition of mycelial growth of both pathogens in vitro. We isolated a total of 32 strains of which 15 came from wild strawberry and 24 commercial strawberry. Only nine strains were obtained with biocontrol potential for one or both pathogens. The highest percentages of mycelial growth inhibition ranged from 67.1% and 81.7% for Botrytis cinerea and 45.5% to 73.2% for Rhizopus stolonifer. These were obtained from four isolates two of them from wild strawberry and the others from commercial strawberry, all with ability to control both pathogens.

  11. Screening of Antagonistic Strain Against Botrytis cinerea%番茄灰霉病菌颉颃菌的筛选

    Institute of Scientific and Technical Information of China (English)

    张雪辉

    2011-01-01

    68 strains were collected fiom different environments around Xingtai University and purified. 16 strains having antagonistics effect to Botrytis cinerea, which account for 23.5% in total were obtained. Among them, 9 strains having great intensive repression to Botrytis cinerea were detected. According to filter-paper detection, the suppression ratio of varieties maintained ranged fiom 65.1% to 92.0%; and bacteria-resistance region ranged fiom 2.0 to 11.0 mm. In addition, 8 anti-bacteria strains were obtained for biological control.%对从不同生态环境下采集的样品进行分离纯化,共得到菌株68株.经初筛,得到对番茄灰霉有颉颃作用的生防菌株16株,占分离菌株的23.5%.并对其中较强颉颃作用的9株菌株进行抑茵活性的测定.结果表明:滤纸片法得到的各菌株对番茄灰霉的抑制率在65.1%~92.0%之间,抑菌带在2.0~11.0mm之间,共获得颉颃菌株8株,占分离菌株的11.8%.

  12. Enhanced resistance to Botrytis cinerea and Rhizoctonia solani in transgenic broccoli with a Trichoderma viride endochitinase gene

    Institute of Scientific and Technical Information of China (English)

    YU Ya; REN Shu-xin; GUO Yang-dong; ZHANG Lei; LIAN Wei-ran; XU Feng-feng; LI Shuang-tao; XIANG Juan; ZHANG Guo-zhen; HU Zan-min; ZHAO Bing

    2015-01-01

    A endochitinase gene (Tch ) from the fungus Trichoderma viride was introduced into broccoli (Brassica oleracea var. italica) by Agrobacterium-mediated transformation. Sixty-eight putative transformants were obtained and the presence of the Tch gene was conifrmed by both PCR and Southern blot analysis. RT-PCR analysis showed an accumulation of the transcript encoding the endochitinase protein in the transgenic plants. Using real-time quantitative PCR, the expression proifling of endochitinase gene was analyzed. Primary transformants and selfed progeny were examined for expression of the endo-chitinase using a lfuorometric assay and for their resistance to the pathogenic fungi Botrytis cinerea and Rhizoctonia solani. The endochitinase activities in T0 in vitro plants, T0 mature plants and T1 mature plants were correlated with leaf lesions, and the transgenic line T618 had high endochitinse activities of 102.68, 114.53 and 120.27 nmol L–1 MU min–1 mg–1 protein in the three kinds of plants, respectively. The endochitinase activity showed a positive correlation with the resistance to the pathogens. Most transgenic T0 broccoli had increased resistance to the pathogens of B. cinerea and R. solani in leaf assays and this resistance was conifrmed to be inheritable. These ifndings suggested that expression of the Tch gene from T. viride could enhance resistance to pathogenic fungi in Brassica species.

  13. A Mechanistic Model of Botrytis cinerea on Grapevines That Includes Weather, Vine Growth Stage, and the Main Infection Pathways.

    Directory of Open Access Journals (Sweden)

    Elisa González-Domínguez

    Full Text Available A mechanistic model for Botrytis cinerea on grapevine was developed. The model, which accounts for conidia production on various inoculum sources and for multiple infection pathways, considers two infection periods. During the first period ("inflorescences clearly visible" to "berries groat-sized", the model calculates: i infection severity on inflorescences and young clusters caused by conidia (SEV1. During the second period ("majority of berries touching" to "berries ripe for harvest", the model calculates: ii infection severity of ripening berries by conidia (SEV2; and iii severity of berry-to-berry infection caused by mycelium (SEV3. The model was validated in 21 epidemics (vineyard × year combinations between 2009 and 2014 in Italy and France. A discriminant function analysis (DFA was used to: i evaluate the ability of the model to predict mild, intermediate, and severe epidemics; and ii assess how SEV1, SEV2, and SEV3 contribute to epidemics. The model correctly classified the severity of 17 of 21 epidemics. Results from DFA were also used to calculate the daily probabilities that an ongoing epidemic would be mild, intermediate, or severe. SEV1 was the most influential variable in discriminating between mild and intermediate epidemics, whereas SEV2 and SEV3 were relevant for discriminating between intermediate and severe epidemics. The model represents an improvement of previous B. cinerea models in viticulture and could be useful for making decisions about Botrytis bunch rot control.

  14. Effects of fludioxonil and pyrimethanil, two fungicides used against Botrytis cinerea, on carbohydrate physiology in Vitis vinifera L.

    Science.gov (United States)

    Saladin, Gaëlle; Magné, Christian; Clément, Christophe

    2003-10-01

    In Vitis vinifera L, photosynthesis and photosynthate partitioning are affected in the presence of fludioxonil and pyrimethanil, two fungicides commonly used in vineyards against Botrytis cinerea Pers. However, the effects were found to be different according to the model studied: plantlets (cv Chardonnay) grown in vitro, fruiting cuttings (cv Chardonnay) and plants grown in vineyards (cvs Chardonnay, Pinot noir and Pinot Meunier). In the plantlets grown in vitro, both fungicides decreased gas exchanges, photosynthetic pigment and starch concentrations in the leaves, whereas soluble carbohydrates transiently accumulated, suggesting that plantlets mobilised starch in response to photosynthesis inhibition caused by fungicides. In the fruiting cuttings, the fungicides did not affect photosynthesis, although fludioxonil caused starch decrease in parallel with sucrose accumulation, suggesting that the fungicide effects were of lower intensity than in vitro. Conversely, in vineyard, the two fungicides stimulated photosynthesis and increased pigment concentrations in the three vine cultivars tested. In the meantime, glucose, fructose and starch levels of the leaves declined after fungicide exposure, whereas sucrose accumulated, indicating that sucrose synthesis increased in the leaves following the fungicide treatment. Among the three varieties, Chardonnay was the most sensitive to the fungicides as revealed by the intensity of the responses and the longer period for recovery. In vineyard, the results suggested that the two fungicides, in addition to inhibiting B cinerea development, had a beneficial effect on vine physiology through the stimulation of leaf carbon nutrition, which may further enable the plant to rapidly make use of its defence reactions.

  15. Testing of Eight Medicinal Plant Extracts in Combination with Kresoxim-Methyl for Integrated Control of Botrytis cinerea in Apples

    Directory of Open Access Journals (Sweden)

    Burtram C. Fielding

    2015-07-01

    Full Text Available Botrytis cinerea is a fungus that causes gray mold on many fruit crops. Despite the availability of a large number of botryticides, the chemical control of gray mold has been hindered by the emergence of resistant strains. In this paper, tests were done to determine the botryticidal efficacy of selected plant extracts alone or combined with kresoxim-methyl. In total, eight South African medicinal plants viz Artemisia afra, Elyptropappus rhinocerotis, Galenia africana, Hypoxis hemerocallidea, Siphonochilus aetheopicus, Sutherlandia frutescens, Tulbaghia violacea and Tulbaghia alliacea were screened. Allium sativum, a plant species known to have antifungal activity, was included in the in vivo studies. For the in vitro studies, synergistic interactions between the plant extracts and the kresoxim-methyl fungicide were tested with radial growth assays. Data indicated synergistic inhibitory effects between the fungicide and the plant extracts. Next, different doses of plant extracts combined with kresoxim-methyl were used for decay inhibition studies on Granny Smith apples. Synergistic and additive effects were observed for many of the combinations. Even though this study was done using only one strain of B. cinerea, results showed that the tested indigenous South African plant species possess natural compounds that potentiate the activity of kresoxim-methyl.

  16. Multidrug resistance in Botrytis cinerea associated with decreased accumulation of the azole fungicide oxpoconazole and increased transcription of the ABC transporter gene BcatrD

    NARCIS (Netherlands)

    Hayashi, K.; Schoonbeek, H.; Sugiura, H.; Waard, De M.A.

    2001-01-01

    Azole-resistant mutants of Botrytis cinerea have a multidrug resistance phenotype since they exhibit cross-resistance to unrelated chemicals. These mutants also display resistance to the new azole fungicide oxpoconazole. Resistance to oxpoconazole is associated with decreased accumulation of the fun

  17. Identification of metabolic pathways expressed by Pichia anomala Kh6 in the presence of the pathogen Botrytis cinerea on apple: new possible targets for biocontrol improvement.

    Science.gov (United States)

    Kwasiborski, Anthony; Bajji, Mohammed; Renaut, Jenny; Delaplace, Pierre; Jijakli, M Haissam

    2014-01-01

    Yeast Pichia anomala strain Kh6 Kurtzman (Saccharomycetales: Endomycetaceae) exhibits biological control properties that provide an alternative to the chemical fungicides currently used by fruit or vegetable producers against main post-harvest pathogens, such as Botrytis cinerea (Helotiales: Sclerotiniaceae). Using an in situ model that takes into account interactions between organisms and a proteomic approach, we aimed to identify P. anomala metabolic pathways influenced by the presence of B. cinerea. A total of 105 and 60 P. anomala proteins were differentially represented in the exponential and stationary growth phases, respectively. In the exponential phase and in the presence of B. cinerea, the pentose phosphate pathway seems to be enhanced and would provide P. anomala with the needed nucleic acids and energy for the wound colonisation. In the stationary phase, P. anomala would use alcoholic fermentation both in the absence and presence of the pathogen. These results would suggest that the competitive colonisation of apple wounds could be implicated in the mode of action of P. anomala against B. cinerea.

  18. 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...

  19. Efecto fungistático de extractos y aceites esenciales de Lippia origanoides HBK y Thymus vulgaris L. como alternativas de manejo de Botrytis cinerea en fresa

    Directory of Open Access Journals (Sweden)

    Luis Alejandro Taborda Andrade

    2015-01-01

    Full Text Available El moho gris de la fresa causado por Botrytis cinerea es una enfermedad que produce importantes pérdidas poscosecha. En el estudio se evaluó el efecto fungistático de extractos y aceites esenciales de Lippia origanoides HBK y Thymus vulgaris L. en concentraciones de 128, 256 y 500 mg/lt sobre B. cinerea in vitro e in vivo. In vitro se determinó el porcentaje de inhibición del crecimiento micelial del hongo. En estas condiciones se observó que el aceite esencial (AE de L. origanoides presentó el porcentaje de control más alto (66.2% sobre B. cinerea. In vivo, se observó que en bananos inoculados con B. cinerea después de 120 los AE controlaron eficientemente la incidencia de daño causado por el patógeno estudiado y no se encontraron diferencias significativas con el control químico utilizando el fungicida Benomil

  20. Bcmfs1, a novel major facilitator superfamily transporter from Botrytis cinerea, provides tolerance towards the natural toxic compounds camptothecin and cercosporin and towards fungicides

    NARCIS (Netherlands)

    Hayashi, K.; Schoonbeek, H.; Waard, De M.A.

    2002-01-01

    Bcmfs1, a novel major facilitator superfamily gene from Botrytis cinerea, was cloned, and replacement and overexpression mutants were constructed to study its function. Replacement mutants showed increased sensitivity to the natural toxic compounds camptothecin and cercosporin, produced by the plant

  1. Identification of metabolic pathways expressed by Pichia anomala Kh6 in the presence of the pathogen Botrytis cinerea on apple: new possible targets for biocontrol improvement.

    Directory of Open Access Journals (Sweden)

    Anthony Kwasiborski

    Full Text Available Yeast Pichia anomala strain Kh6 Kurtzman (Saccharomycetales: Endomycetaceae exhibits biological control properties that provide an alternative to the chemical fungicides currently used by fruit or vegetable producers against main post-harvest pathogens, such as Botrytis cinerea (Helotiales: Sclerotiniaceae. Using an in situ model that takes into account interactions between organisms and a proteomic approach, we aimed to identify P. anomala metabolic pathways influenced by the presence of B. cinerea. A total of 105 and 60 P. anomala proteins were differentially represented in the exponential and stationary growth phases, respectively. In the exponential phase and in the presence of B. cinerea, the pentose phosphate pathway seems to be enhanced and would provide P. anomala with the needed nucleic acids and energy for the wound colonisation. In the stationary phase, P. anomala would use alcoholic fermentation both in the absence and presence of the pathogen. These results would suggest that the competitive colonisation of apple wounds could be implicated in the mode of action of P. anomala against B. cinerea.

  2. Production and Accumulation of Xylooligosaccharides with Long Chains by Growing Culture and Xylanase of a Mutant Strain of Bacillus pumilus X-6-19

    Institute of Scientific and Technical Information of China (English)

    Qingzhu Yuan; Tsuyoshi Adachi; Shinji Takenaka; Shuichiro Murakami; Machiko Tanaka; Kenji Aoki

    2008-01-01

    Bacillus pumilus X-6-9 isolated from soil and subsequently identified, produced xylooligosacchatides with long chainsfrom xylan and accumulated them in the culture. By improving the culture conditions and mutating the bacterium, a 3.2-fold increasein the production of the xylooligosaccharides was established, when compared to the original culture conditions of B. pumilus X-6-19.The addition of D-glucose to the culture of the mutant swain U-3 of B. pumilus X-6-9 repressed the synthesis of β-xylosidase, but notxylanase. Thus, it was revealed that strain U-3 was a good organism for the production and accumulation of xylooligosaccharideswith long chains from xylan by a microbial culture. Xylanase produced by strain U-3 was purified to homogeneity and characterized.The hydrolyzates generated by the purified xylanase contained xylobiose, xylotrinse, xylotewaose, and xylopentaose, but not xylose.

  3. The N-terminal cellulose-binding domain of EGXA increases thermal stability of xylanase and changes its specific activities on different substrates

    Institute of Scientific and Technical Information of China (English)

    Ming Ding; Yigang Teng; Qiuyu Yin; Jie Zhao; Fukun Zhao

    2008-01-01

    A full-length EGXA enzyme from a mollusk, Ampullaria crossean, was cloned into pFastBac vector and then heterogeneously expressed in insect Tn5 cells. Its natural N-terminal signal peptide worked well in the insect Tn5 cells.The recombinant EGXA was a 63 kDa protein and had active endo-β-1,4-glucanase (EC 3.2.1.4) and endo-β-1,4-xylanase (EC 3.2.1.8). The specific activity of endo-β-1,4-xylanase was higher than in the EGX, which was purified from the stomach tissues of Ampullaria crossen. The N-terminal cellulosebinding domain of EGXA made it bind to cellulose and xylan more efficiently. This cellulose-binding domain also increased the thermal stability of this recombinant enzyme and decreased the recombinant EGXA's specific activities on p-nitrophenyi-β-D-cellobioside and sodium carboxymethyl cellulose.

  4. Individual and combined effects of water addition with xylanases and laccase on the loaf quality of composite wheat–cassava bread

    DEFF Research Database (Denmark)

    Serventi, Luca; Skibsted, Leif H.; Kidmose, Ulla

    2016-01-01

    hardness was measured at 58 % water addition, likely due to insufficient plasticization of the gluten–starch network, while hardening and crumb structure collapse were observed at 76 % water addition. Enzyme evaluation revealed higher pore size upon treatment with the xylanase Panzea® BG (Panzea) compared......The objective was to study how water addition and addition of enzymes like xylanases and a laccase will improve the loaf quality of composite wheat–cassava bread. The loaf quality was determined by sensory profiling, volume measurement and texture profile analysis. High intensity of sensory...... of polyphenols and/or arabinoxylans. In confirmation to these findings, the combination of high water addition (70 %) and Panzea treatment generated larger and better structured bread....

  5. Effect of Different Pretreatment of Sugar Cane Bagasse on Cellulase and Xylanases Production by the Mutant Penicillium echinulatum 9A02S1 Grown in Submerged Culture

    OpenAIRE

    Marli Camassola; Dillon, Aldo J.P.

    2014-01-01

    The main limitation to the industrial scale hydrolysis of cellulose is the cost of cellulase production. This study evaluated cellulase and xylanase enzyme production by the cellulolytic mutant Penicillium echinulatum 9A02S1 using pretreated sugar cane bagasse as a carbon source. Most cultures grown with pretreated bagasse showed similar enzymatic activities to or higher enzymatic activities than cultures grown with cellulose or untreated sugar cane bagasse. Higher filter paper activity (1.25...

  6. Ectoine-mediated protection of enzyme from the effect of pH and temperature stress: a study using Bacillus halodurans xylanase as a model.

    Science.gov (United States)

    Van-Thuoc, Doan; Hashim, Suhaila O; Hatti-Kaul, Rajni; Mamo, Gashaw

    2013-07-01

    Compatible solutes are small, soluble organic compounds that have the ability to stabilise proteins against various stress conditions. In this study, the protective effect of ectoines against pH stress is examined using a recombinant xylanase from Bacillus halodurans as a model. Ectoines improved the enzyme stability at low (4.5 and 5.0) and high pH (11 and 12); stabilisation effect of hydroxyectoine was superior to that of ectoine and trehalose. In the presence of hydroxyectoine, residual activity (after 10 h heating at 50 °C) increased from about 45 to 86 % at pH 5 and from 33 to 89 % at pH 12. When the xylanase was incubated at 65 °C for 5 h with 50 mM hydroxyectoine at pH 10, about 40 % of the original activity was retained while no residual activity was detected in the absence of additives or in the presence of ectoine or trehalose. The xylanase activity was slightly stimulated in the presence of 25 mM ectoines and then gradually decreased with increase in ectoines concentration. The thermal unfolding of the enzyme in the presence of the compatible solutes showed a modest increase in denaturation temperature but a larger increase in calorimetric enthalpy.

  7. Parthenium sp. as a plant biomass for the production of alkalitolerant xylanase from mutant Penicillium oxalicum SAU{sub E}-3.510 in submerged fermentation

    Energy Technology Data Exchange (ETDEWEB)

    Dwivedi, Pallavi; Vivekanand, V.; Ganguly, Ruma; Singh, Rajesh P. [Department of Biotechnology, Indian Institute of Technology Roorkee, Roorkee 247667 (India)

    2009-04-15

    The use of congress grass (Parthenium sp.) and water hyacinth (Eichhornia crassipes) as low cost raw materials for xylanase production from mutant Penicillium oxalicum SAU{sub E}-3.510 in submerged fermentation was investigated. For development of mutant from wild type P. oxalicum SA-8 ITCC 6024, a strategy of mixed mutagenesis was followed using UV-irradiation and ethidium bromide, which had resulted into 1.87 fold increases in the activity of the enzyme. For enzyme production, the fungus was cultivated in mineral medium containing congress grass as carbon source. Considerably higher levels of production (475.2 {+-} 6.0 IU ml{sup -1}) were achieved in media containing congress grass, although it was slightly less than that was obtained (488.5 {+-} 6.5 IU ml{sup -1}) in presence of commercial oat spelt xylan. This fact confirms the feasibility of using this low cost non-food resource as an alternative carbon source to save costs of the enzyme production process. Maximum xylanase activity was reported at 55 C with its stability at 80 C for 2 h. The highest activity of xylanase at pH 9.0 and its stability at similar pH for 24 h denote the alkalitolerant nature of enzyme. (author)

  8. Supplementation with xylanase and β-xylosidase to reduce xylo-oligomer and xylan inhibition of enzymatic hydrolysis of cellulose and pretreated corn stover

    Directory of Open Access Journals (Sweden)

    Qing Qing

    2011-06-01

    Full Text Available Abstract Background Hemicellulose is often credited with being one of the important physical barriers to enzymatic hydrolysis of cellulose, and acts by blocking enzyme access to the cellulose surface. In addition, our recent research has suggested that hemicelluloses, particularly in the form of xylan and its oligomers, can more strongly inhibit cellulase activity than do glucose and cellobiose. Removal of hemicelluloses or elimination of their negative effects can therefore become especially pivotal to achieving higher cellulose conversion with lower enzyme doses. Results In this study, cellulase was supplemented with xylanase and β-xylosidase to boost conversion of both cellulose and hemicellulose in pretreated biomass through conversion of xylan and xylo-oligomers to the less inhibitory xylose. Although addition of xylanase and β-xylosidase did not necessarily enhance Avicel hydrolysis, glucan conversions increased by 27% and 8% for corn stover pretreated with ammonia fiber expansion (AFEX and dilute acid, respectively. In addition, adding hemicellulase several hours before adding cellulase was more beneficial than later addition, possibly as a result of a higher adsorption affinity of cellulase and xylanase to xylan than glucan. Conclusions This key finding elucidates a possible mechanism for cellulase inhibition by xylan and xylo-oligomers and emphasizes the need to optimize the enzyme formulation for each pretreated substrate. More research is needed to identify advanced enzyme systems designed to hydrolyze different substrates with maximum overall enzyme efficacy.

  9. [Cultivation of a novel cellulase/xylanase producer, Trichoderma longibrachiatum mutant TW 1-59-27: production of the enzyme preparation and the study of its properties].

    Science.gov (United States)

    Bekkarevicha, A O; Nemashkalov, V A; Koshelev, A V; Goryazchev, D A; Bubnova, T V; Matys, V Yu; Osipov, D O; Kondrat'eva, E G; Okunev, O N; Sinitsyn, A P

    2015-01-01

    As a result of gamma-mutagenesis of Trichoderma longibrachiatum TW1 and the subsequent selection of improved producers, a novel mutant strain, TW1-59-27, capable of efficiently secreting cellulase and xylanase was obtained. In a fed-batch cultivation, the new TW1-59-27 mutant was significantly more active compared with the original TW1 strain. For instance, the activities of cellulase (towards carboxymethylcellulose) and xylanase in the culture broth (CB) increased by 1.8 and two times, respectively, and the protein content increased by 1.47 times. The activity of these enzymes in the dry enzyme preparation derived from the CB of the TW1-59-27 mutant was 1.3-1.8 times higher than that in the preparation derived from the original TW1 strain. It was established that the cellulase from the enzyme preparation of the mutant strain demonstrated the maximum activity at 55-65 degrees C; it occurred in xylanase at 60 degrees C. The pH optima of these enzymes were pH 4.5-5.0 and pH 5.0-6.0, respectively. It was shown that the content of endoglucanases in the enzyme preparation increased from 7% to 13.5%; the effect is largely driven by the secretion of endoglucanase-1. An enzyme preparation with increased endoglucanase-1 content is promising for use as a feed additive in agriculture.

  10. An approach to the production of soluble protein from a fungal gene encoding an aggregation-prone xylanase in Escherichia coli.

    Science.gov (United States)

    Le, Yilin; Peng, Jingjing; Wu, Huawei; Sun, Jianzhong; Shao, Weilan

    2011-04-08

    The development of new procedures and protocols that allow researchers to obtain recombinant proteins is of fundamental importance in the biotechnology field. A strategy was explored to overcome inclusion-body formation observed when expressing an aggregation-prone fungal xylanase in Escherichia coli. pHsh is an expression plasmid that uses a synthetic heat-shock (Hsh) promoter, in which gene expression is regulated by an alternative sigma factor (σ(32)). A derivative of pHsh was constructed by fusing a signal peptide to xynA2 gene to facilitate export of the recombinant protein to the periplasm. The xylanase was produced in a soluble form. Three factors were essential to achieving such soluble expression of the xylanase: 1) the target gene was under the control of the Hsh promoter, 2) the gene product was exported into the periplasm, and 3) gene expression was induced by a temperature upshift. For the first time we report the expression of periplasmic proteins under the control of an Hsh promoter regulated by σ(32). One unique feature of this approach was that over 200 copies of the Hsh promoter in an E. coli cell significantly increased the concentration of σ(32). The growth inhibition of the recombinant cells corresponded to an increase in the levels of soluble periplasmic protein. Therefore, an alternative protocol was designed to induce gene expression from pHsh-ex to obtain high levels of active soluble enzymes.

  11. Study on xylanase production by Neurospora sitophila in liquid fermentation%好食脉孢霉液态发酵产木聚糖酶的研究

    Institute of Scientific and Technical Information of China (English)

    邓永平; 刘晓兰; 艾瑞波; 郑喜群; 任凭

    2013-01-01

    试验以豆渣和麸皮为主要原料,研究了好食脉孢霉菌株产木聚糖酶的液态发酵条件.以木聚糖酶的催化活力为指标,确定液态发酵培养基由麸皮、豆渣和蛋白胨组成,初始pH值5.0、培养温度28℃、摇床转速150 r/min,培养72 h后木聚糖酶活力可达到476 U/ml.%Okara and bran as the main raw material, the liquid fermentation conditions of xylanase production by Neurospora sitophila was studied in this paper. Using xylanase catalytic activity as an indicator, the compositions of the liquid fermentation medium which were bran, okara and peptone were determined. When the initial pH was 5.0, incubation temperature was 28 ℃, the shaking speed was 150 r/min and culture time was 72 h, xylanase activity was 476 U/ml.

  12. Identification and Characterization of Botrytis Blossom Blight of Japanese Plums Caused by Botrytis cinerea and B. prunorum sp. nov. in Chile.

    Science.gov (United States)

    Ferrada, Enrique E; Latorre, Bernardo A; Zoffoli, Juan P; Castillo, Antonio

    2016-02-01

    Blossom blight is a destructive disease of plums (Prunus salicina) when humid and temperate weather conditions occur in Chile. Disease incidence ranging from 4 to 53% has been observed. Symptoms include light brown petal necrosis, starting as light brown mottles or V-shaped necrosis at the margins of the petals, progressing to the stamen and pistils. In this study, the etiology of blossom blight of plums was determined. High- and low-sporulating isolates of Botrytis were obtained consistently from blighted blossoms and apparently healthy flowers of plums. Based on colony morphology, conidial production and molecular phylogenetic analysis, these high- and low-sporulating isolates were identified as B. cinerea and B. prunorum sp. nov., respectively. Phylogenetic analysis of the genes glyceraldehyde 3-phosphate dehydrogenase (G3PDH), heat-shock protein 60 (HSP60), and DNA-dependent RNA polymerase subunit II (RPB2) grouped B. prunorum isolates in a single cluster, distantly from B. cinerea and other Botrytis species. The phylogenetic analysis of necrosis and ethylene-inducing protein (NEP1 and NEP2) genes corroborated these results. Analysis of the internal transcribed spacer region and large-subunit (26S) ribosomal DNA and detection of Boty and Flipper transposable elements, were not useful to differentiate between these Botrytis species. Both species were pathogenic on plum flowers and the fruit of plums, apples, and kiwifruits. However, B. prunorum was less virulent than B. cinerea. These pathogens were re-isolated from inoculated and diseased tissues; thus, Koch's postulates were fulfilled, confirming its role in blossom blight of plums. B. cinerea was predominant, suggesting that B. prunorum may play a secondary role in the epidemiology of blossom blight in plums in Chile. This study clearly demonstrated that the etiology of blossom blight of plums is caused by B. cinerea and B. prunorum, which constitute a species complex living in sympatry on plums and possibly

  13. 紫皮石斛上灰葡萄孢的分离和鉴定%Isolation and Identification of Botrytis cinerea from Dendrobium devonianum

    Institute of Scientific and Technical Information of China (English)

    赵菊润; 席刚俊; 赵桂华

    2014-01-01

    For understanding the relationship between Dendrobium devonianum and fungal on it , tissue isolation were conducted for root , stem and leaf in both diseased and healthy Dendrobium devonianum, and a total 306 fun-gal strains were obtained .Among these fungal strains , the largest number was Botrytis cinerea, which accounted for 23.4 %. After purified culturing , the Botrytis cinerea were identified by means of combining morphological characters with molecular biological techniques .Based on the the fact of host range and distribution range of Botry-tis cinerea, this study showed that D.devonianum is a new host for Botrytis cinerea and this was the first report that D.devonianum could be host of Botrytis cinerea.%为了研究紫皮石斛上的真菌,对紫皮石斛染病和健康根、茎和叶片进行了组织分离,共得到306个真菌菌株,其中灰葡萄孢出现数量最多,占23.4%。通过纯化培养,并采用形态特征和分子生物技术相结合的方法对灰葡萄孢进行了鉴定。根据该菌的寄主范围和分布地区,结果表明,紫皮石斛是灰葡萄孢的新寄主,此菌在紫皮石斛上发生属于首次报道。

  14. Involvement of protein tyrosine phosphatases BcPtpA and BcPtpB in regulation of vegetative development, virulence and multi-stress tolerance in Botrytis cinerea.

    Directory of Open Access Journals (Sweden)

    Qianqian Yang

    Full Text Available Tyrosine phosphorylation and dephosphorylation have emerged as fundamentally important mechanisms of signal transduction and regulation in eukaryotic cells, governing many processes, but little has been known about their functions in filamentous fungi. In this study, we deleted two putative protein tyrosine phosphatase (PTP genes (BcPTPA and BcPTPB in Botrytis cinerea, encoding the orthologs of Saccharomyces cerevisiae Ptp2 and Ptp3, respectively. Although BcPtpA and BcPtpB have opposite functions in conidiation, they are essential for sclerotial formation in B. cinerea. BcPTPA and BcPTPB deletion mutants ΔBcPtpA-10 and ΔBcPtpB-4 showed significantly increased sensitivity to osmotic and oxidative stresses, and to cell wall damaging agents. Inoculation tests showed that both mutants exhibited dramatically decreased virulence on tomato leaves, apples and grapes. In S. cerevisiae, it has been shown that Ptp2 and Ptp3 negatively regulate the high-osmolarity glycerol (HOG pathway and the cell wall integrity (CWI pathway. Although both BcPtpA and BcPtpB were able to inactive Hog1 and Mpk1 in S. cerevisiae, in contrast to S. cerevisiae, they positively regulate phosphorylation of BcSak1 (the homologue of Hog1 and BcBmp3 (the homologue of Mpk1 in B. cinerea under stress conditions. These results demonstrated that functions of PTPs in B. cinerea are different from those in S. cerevisiae, and BcPtpA and BcPtpB play important roles in regulation of vegetative development, virulence and in adaptation to oxidative, osmotic and cell-wall damage stresses in B. cinerea.

  15. Interactive effects of phytase and xylanase supplementation with extractable salt-soluble protein content of corn in diets with adequate calcium and nonphytate phosphorus fed to broilers.

    Science.gov (United States)

    Gehring, C K; Bedford, M R; Dozier, W A

    2013-07-01

    The objective was to determine the effects of extractable salt-soluble protein content of corn (PS) and exogenous enzyme supplementation on N, starch, and energy digestibility in broilers fed diets adequate in Ca and nonphytate P. Broilers were randomly distributed into floor pens (6 replicate pens per treatment) with 28 birds per pen at 1 d of age. Treatments consisting of 4 sources of corn varying in PS (A, 58.1; B, 54.2; C, 53.7; and D, 30.6 mg of BSA equivalent values) with or without phytase (0 and 1,000 phytase units/kg) and xylanase (0 and 16,000 units of xylanase activity/kg) were randomly assigned to each pen. Different sources of corn were provided from 1 to 9 and 24 to 29 d of age. However, enzyme treatments were provided throughout the experiment. From 1 to 9 d of age, no interactions were observed. Apparent ileal N digestibility (AIND) and apparent ileal digestible energy (IDE) of diets with the lowest PS (based on corn D) were lower (P ≤ 0.05) than those of diets with a higher PS. Phytase increased (P ≤ 0.01) AIND and IDE by 5 and 16%, respectively, and xylanase exerted the opposite effect (P ≤ 0.03). From 24 to 29 d of age, phytase and xylanase in combination resulted in reduced (P ≤ 0.05) AIND of diets with a low PS (based on corn D) compared with the basal diet in broilers. Broilers fed diets with the highest or lowest PS (based on corn A or D) had lower (3-way interaction; P ≤ 0.05) IDE when phytase and xylanase were supplemented in combination compared with either enzyme alone. In conclusion, responses to exogenous enzyme supplementation are not constant and are influenced by the source of ingredients as well as the age of broilers. The magnitudes of the responses to phytase on nutrient and energy digestibility were greater at 9 compared with 29 d of age.

  16. Cloning and expression of the xynA1 gene encoding a xylanase of the GH10 group in Caulobacter crescentus.

    Science.gov (United States)

    Graciano, Luciana; Corrêa, Juliana Moço; Vieira, Fabíola Giovanna Nesello; Bosetto, Adilson; Loth, Eduardo Alexandre; Kadowaki, Marina Kimiko; Gandra, Rinaldo Ferreira; Simão, Rita de Cássia Garcia

    2015-04-01

    Caulobacter crescentus (NA1000 strain) are aquatic bacteria that can live in environments of low nutritional quality and present numerous genes that encode enzymes involved in plant cell wall deconstruction, including five genes for β-xylosidases (xynB1-xynB5) and three genes for xylanases (xynA1-xynA3). The overall activity of xylanases in the presence of different agro-industrial residues was evaluated, and it was found that the residues from the processing of corn were the most efficient in inducing bacterial xylanases. The xynA1 gene (CCNA_02894) encoding a predicted xylanase of group 10 of glyco-hydrolases (GH10) that was efficiently overexpressed in Escherichia coli LMG194 using 0.02 % arabinose, after cloning into the vector pJet1.2blunt and subcloning into the expression vector pBAD/gIII, provided a fusion protein that contained carboxy-terminal His-tags, named XynA1. The characterization of pure XynA1 showed an enzymatic activity of 18.26 U mL(-1) and a specific activity of 2.22 U mg-(1) in the presence of xylan from beechwood as a substrate. XynA1 activity was inhibited by EDTA and metal ions such as Cu(2+) and Mg(2+). By contrast, β-mercaptoethanol, dithiothreitol (DTT), and Ca(2+) induced recombinant enzyme activity. Kinetic data for XynA1 revealed K M and V max values of 3.77 mg mL-(1) and 10.20 μM min-(1), respectively. Finally, the enzyme presented an optimum pH of 6 and an optimum temperature of 50 °C. In addition, 80 % of the activity of XynA1 was maintained at 50 °C for 4 h of incubation, suggesting a thermal stability for the biotechnological processes. This work is the first study concerning the cloning, overexpression, and enzymatic characterization of C. crescentus xylanase.

  17. Study on antifungal activities of biocontrol yeasts against Botrytis cinerea%三株生防酵母菌对Botrytis cinerea的抑菌作用研究

    Institute of Scientific and Technical Information of China (English)

    王傲雪; 关鑫; 张俊峰; 张珍珠; 王瑞虎; 陈秀玲

    2014-01-01

    In this study, the antifungal activities of biocontrol yeasts Cryptococcus albidus 63(Ca63), Cryptococcus albidus 64 (Ca64) and Candida parapsilosis yett1006 against Botrytis cinerea were investigated. The dual culture between three biocontrol yeasts and B. cinerea t08016b results showed that no obvious inhibiting zone appeared, but the pathogen growed more weakly than the control. The results of chitinase andβ-1, 3-glucanase activities assay showed that the levels of the two enzymes activities did not change significantly (P>0.05) when the three biocontrol yeasts cultured with pathogen. The inhibition rates of three biocontrol yeast strains on mycelium growth were 88.57%, 90.66% and 85.65%, respectively. High concentrations of biocontrol yeasts had a strong inhibitory effect on the spore germination of B. cinerea t08016b. Especial y, 1 × 108 cfu·mL-1 biocontrol yeasts almost stopped the spore germination of B. cinerea t08016b. Heat-inactived yeast suspension and culture supernatant had no significant inhibitory effects on B. cinerea. These results il ustrated that the antagonic effect of biocontrol yeasts was mainly due to the competition of limited space and nutrient conditions, not secreting antifungal substances.%针对三株生防酵母菌Cryptococcus albidus 63(Ca63)、Cryptococcus albidus 64(Ca64)、Candida parapsilosis yett1006对番茄灰霉病病原菌B. cinerea t08016b的抑菌作用进行研究。三株生防酵母菌与B. cinerea t08016b的对峙培养过程中,未产生明显抑菌圈,均可减弱B. cinerea t08016b生长势;几丁质酶和β-1,3-葡聚糖酶活性测定结果表明, t08016b并无诱导三株酵母菌分泌酶活显著增加(P>0.05);研究酵母菌对B. cinerea t08016b菌丝生长和孢子萌发率影响发现,C.albidus 63、C.albidus 64和C.parapsilosis yett1006对菌丝生长量的抑制率分别为88.57%、90.66%和85.65%,高浓度菌液对孢子萌发有显著抑制效果,1×108 cfu·mL-1菌体活

  18. Trophic ecology of the Grey Wagtail Motacilla cinerea before and during the breeding season in the region of Bejaia (Algeria).

    Science.gov (United States)

    Bougaham, Abdelazize Franck; Moulaï, Riadh; O'Halloran, John

    2014-01-01

    The diet composition of the Grey Wagtail Motacilla cinerea of the Babor Range is followed by analysis of faecal samples (90 faeces) before and during the breeding season 2010. The Grey Wagtail's diet varies depending on the stage of the breeding cycle at the southern edge of their breeding area in North Africa (Bejaia). The diet consists predominantly of aquatic preys (51.79%), with Coleoptera being the most frequent constituent (n=331, 45.5%). During the pre-laying period (February-March), the diet was variable (91 prey-taxa and H'=3,36 bits) and preys of medium size (5 to 8mm) were most common. During the incubation period (April-May), preys were mainly aquatic (60%) and larger (20 to 32mm). At the end of the breeding season (June-July), there was a greater occurrence of terrestrial preys (31 aquatic versus 30 terrestrial taxa).

  19. Inhibitory effect of Xenorhabdus nematophila TB on plant pathogens Phytophthora capsici and Botrytis cinerea in vitro and in planta.

    Science.gov (United States)

    Fang, Xiangling; Zhang, Manrang; Tang, Qian; Wang, Yonghong; Zhang, Xing

    2014-03-06

    Entomopathogenic bacteria Xenorhabdus spp. produce secondary metabolites with potential antimicrobial activity for use in agricultural productions. This study evaluated the inhibitory effect of X. nematophila TB culture on plant pathogens Botrytis cinerea and Phytophthora capsici. The cell-free filtrate of TB culture showed strong inhibitory effects (>90%) on mycelial growth of both pathogens. The methanol-extracted bioactive compounds (methanol extract) of TB culture also had strong inhibitory effects on mycelial growth and spore germinations of both pathogens. The methanol extract (1000 μg/mL) and cell-free filtrate both showed strong therapeutic and protective effects (>70%) on grey mold both in detached tomato fruits and plants, and leaf scorch in pepper plants. This study demonstrates X. nematophila TB produces antimicrobial metabolites of strong activity on plant pathogens, with great potential for controlling tomato grey mold and pepper leaf scorch and being used in integrated disease control to reduce chemical application.

  20. Isolation and characterization of Bacillus subtilis EB-28, an endophytic bacterium strain displaying biocontrol activity against Botrytis cinerea Pers

    Institute of Scientific and Technical Information of China (English)

    Shutong WANG; Tongle HU; Yanling JIAO; Jianjian WEI; Keqiang CAO

    2009-01-01

    The fungal pathogen Botrytis cinerea Pers. causes severe rotting on tomato fruits during storage and shelf life. As a biological control agent, endophytic bacterium was regarded as an effective alternative to chemical control. Out of 238 endophytic bacterial isolates, three strains (EB-15, EB-28, and EB-122) isolated from Lycopersicum esculentum Mill., Speranskia tuberculata (Bge.) Baill, and Dictamnus dasycarpus Turcz. respectively were found to be strongly antagonistic to the pathogen in vitro and were selected for further in vivo tests. One endophytic bacterium strain, encoded EB-28, was selected from the three in vivo tested isolates. The inhibitive rate of EB-28 reached 71.1% in vitro and 52.4% in vivo. EB-28 was identified as Bacillus subtilis according to its morphological, physiological, and biochemical characteristics and 16S rDNA sequence analysis.