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Sample records for endophytic bacillus pumilus

  1. Genome Sequence of the Plant Endophyte Bacillus pumilus INR7, Triggering Induced Systemic Resistance in Field Crops

    PubMed Central

    Jeong, Haeyoung; Choi, Soo-Keun; Kloepper, Joseph W.

    2014-01-01

    Bacillus pumilus INR7 is an endophytic bacterium that has been commercialized as a biological control product against soilborne pathogens as well as foliar pathogens by direct antagonism and induction of systemic resistance. In the current study, we provide the genome sequence and a possible explanation of the function of strain INR7. PMID:25359912

  2. Bacillus pumilus SAFR-032 isolate

    NASA Technical Reports Server (NTRS)

    Venkateswaran, Kasthuri J. (Inventor)

    2007-01-01

    The present invention relates to discovery and isolation of a biologically pure culture of a Bacillus pumilus SAFR-032 isolate with UV sterilization resistant properties. This novel strain has been characterized on the basis of phenotypic traits, 16S rDNA sequence analysis and DNA-DNA hybridization. According to the results of these analyses, this strain belongs to the genus Bacillus. The GenBank accession number for the 16S rDNA sequence of the Bacillus pumilus SAFR-032 isolate is AY167879.

  3. Complete Genome of Bacillus pumilus Siphophage Glittering

    PubMed Central

    Matthew, Solomon P.; Decker, Skyelar L.; Chamakura, Karthik R.

    2013-01-01

    Bacillus pumilus is a Gram-positive bacterium widely used in agriculture both as an antifungal and as a growth-promoting symbiont. B. pumilus is rarely infectious but has recently been shown to infect humans. Here, we present the complete genome of B. pumilus phage Glittering, a potential biocontrol agent for B. pumilus. PMID:24309725

  4. [Extracellular ribonuclease from Bacillus pumilus].

    PubMed

    Struminskaia, N K; Iva?lovski?, V L; Dement'ev, A A; Moiseev, G P; Fedosov, Iu A; Iakovlev, G I

    1992-01-01

    The extracellular ribonuclease (RNAse Bp) was isolated from the cultural medium filtrate of Bacillus pumilus by ammonium sulfate precipitation and two stages of ion-exchange chromatography on carboxymethyl- and phospho-cellulose columns. The amino acid composition and N-terminal amino acid residue have been determined. The kinetic parameters of cleavage reaction of synthetic polynucleotides have been measured. According to their structural homology RNAse Bp has been shown to be similar to RNAses Ba and Bi. Catalytic properties of the enzyme are very close to RNAse Bi. PMID:1302501

  5. Protection of Bacillus pumilus Spores by Catalases

    PubMed Central

    Checinska, Aleksandra; Burbank, Malcolm

    2012-01-01

    Bacillus pumilus SAFR-032, isolated at spacecraft assembly facilities of the National Aeronautics and Space Administration Jet Propulsion Laboratory, is difficult to kill by the sterilization method of choice, which uses liquid or vapor hydrogen peroxide. We identified two manganese catalases, YjqC and BPUM_1305, in spore protein extracts of several B. pumilus strains by using PAGE and mass spectrometric analyses. While the BPUM_1305 catalase was present in six of the B. pumilus strains tested, YjqC was not detected in ATCC 7061 and BG-B79. Furthermore, both catalases were localized in the spore coat layer along with laccase and superoxide dismutase. Although the initial catalase activity in ATCC 7061 spores was higher, it was less stable over time than the SAFR-032 enzyme. We propose that synergistic activity of YjqC and BPUM_1305, along with other coat oxidoreductases, contributes to the enhanced resistance of B. pumilus spores to hydrogen peroxide. We observed that the product of the catalase reaction, gaseous oxygen, forms expanding vesicles on the spore surface, affecting the mechanical integrity of the coat layer, resulting in aggregation of the spores. The accumulation of oxygen gas and aggregations may play a crucial role in limiting further exposure of Bacilli spore surfaces to hydrogen peroxide or other toxic chemicals when water is present. PMID:22752169

  6. Protection of Bacillus pumilus spores by catalases.

    PubMed

    Checinska, Aleksandra; Burbank, Malcolm; Paszczynski, Andrzej J

    2012-09-01

    Bacillus pumilus SAFR-032, isolated at spacecraft assembly facilities of the National Aeronautics and Space Administration Jet Propulsion Laboratory, is difficult to kill by the sterilization method of choice, which uses liquid or vapor hydrogen peroxide. We identified two manganese catalases, YjqC and BPUM_1305, in spore protein extracts of several B. pumilus strains by using PAGE and mass spectrometric analyses. While the BPUM_1305 catalase was present in six of the B. pumilus strains tested, YjqC was not detected in ATCC 7061 and BG-B79. Furthermore, both catalases were localized in the spore coat layer along with laccase and superoxide dismutase. Although the initial catalase activity in ATCC 7061 spores was higher, it was less stable over time than the SAFR-032 enzyme. We propose that synergistic activity of YjqC and BPUM_1305, along with other coat oxidoreductases, contributes to the enhanced resistance of B. pumilus spores to hydrogen peroxide. We observed that the product of the catalase reaction, gaseous oxygen, forms expanding vesicles on the spore surface, affecting the mechanical integrity of the coat layer, resulting in aggregation of the spores. The accumulation of oxygen gas and aggregations may play a crucial role in limiting further exposure of Bacilli spore surfaces to hydrogen peroxide or other toxic chemicals when water is present. PMID:22752169

  7. Cutaneous infection due to Bacillus pumilus: report of 3 cases.

    PubMed

    Tena, Daniel; Martinez-Torres, Juan Angel; Perez-Pomata, María Teresa; Sáez-Nieto, Juan Antonio; Rubio, Virginia; Bisquert, Julia

    2007-02-15

    Human infection due to Bacillus pumilus is exceptional. We report 3 cases of cutaneous infection caused by B. pumilus that occurred in 3 shepherds, 2 of whom were members of the same family. The lesions appeared to have a morphology similar to that of cutaneous anthrax lesions. Two patients were cured after treatment with amoxicillin-clavulanate, and the third patient was cured after prolonged treatment with ciprofloxacin. To our knowledge, primary cutaneous infection due to B. pumilus has not been reported. B. pumilus should be considered in patients who develop lesions suggestive of cutaneous anthrax. PMID:17243047

  8. A novel glycerophosphodiesterase from Bacillus pumilus.

    PubMed

    Kusser, W; Fiedler, F

    1984-01-30

    A novel glycerophosphodiesterase activity was detected in extracts from phosphate-starved Bacillus pumilus DSM27 cells. The enzyme had a substrate specificity for glycerophosphodiester bonds and the reaction product formed with partially purified enzyme was (sn)-glycero-3-phosphate. Purified cell wall teichoic acid of the polyglycerophosphate type, as well as deacylated, unsubstituted lipoteichoic acid of the polyglycerophosphate type, di(glycerophospho)glycerol (deacylated cardiolipin) and mono(glycerophospho)glycerol (deacylated phosphatidylglycerol) served as substrates for the enzyme. Their native counterparts, however, cell wall-bound polyglycerophosphate, lipoteichoic acid (D-alanine substituted and dealanylated), cardiolipin and phosphatidylglycerol were poor or no substrates, respectively. Enzyme activity was inhibited by purified cell walls and by heparin. The enzyme was partially purified using a column of Heparin-Sepharose. PMID:6319195

  9. Increased radiation resistance of vegetative Bacillus pumilus.

    PubMed

    Parisi, A; Antoine, A D

    1974-07-01

    A 4.5-fold increase in vegetative cell radiation resistance of Bacillus pumilus E601, the internationally recognized biological standard for irradiation sterilization, was obtained by the repeated passage of resistant survivors through successive sublethal doses of (60)Co irradiation. This increase in resistance was accompanied by a corresponding increase in spore resistance through the seventh irradiation passage. By the fifteenth passage, the ability for spore formation was lost. Other effects noted by the successive irradiation dosages included loss of motility and pellicle formation, and changes in the Gram reaction, cell morphology, and colonial morphology. Increased resistance was also accompanied by an increased nutritional requirement for specific amino acids. Radiation resistance was not transferred from vegetative cells to spores. PMID:4844266

  10. 40 CFR 180.1224 - Bacillus pumilus GB34; exemption from the requirement of a tolerance.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 40 Protection of Environment 24 2014-07-01 2014-07-01 false Bacillus pumilus GB34; exemption from... FOOD Exemptions From Tolerances § 180.1224 Bacillus pumilus GB34; exemption from the requirement of a... pesticide Bacillus pumilus GB34 when used as a seed treatment in or on all food commodities. An exemption...

  11. 40 CFR 180.1224 - Bacillus pumilus GB34; exemption from the requirement of a tolerance.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 40 Protection of Environment 24 2011-07-01 2011-07-01 false Bacillus pumilus GB34; exemption from... FOOD Exemptions From Tolerances § 180.1224 Bacillus pumilus GB34; exemption from the requirement of a... pesticide Bacillus pumilus GB34 when used as a seed treatment in or on all food commodities. An exemption...

  12. 40 CFR 180.1224 - Bacillus pumilus GB34; exemption from the requirement of a tolerance.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 40 Protection of Environment 25 2013-07-01 2013-07-01 false Bacillus pumilus GB34; exemption from... FOOD Exemptions From Tolerances § 180.1224 Bacillus pumilus GB34; exemption from the requirement of a... pesticide Bacillus pumilus GB34 when used as a seed treatment in or on all food commodities. An exemption...

  13. 40 CFR 180.1224 - Bacillus pumilus GB34; exemption from the requirement of a tolerance.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 23 2010-07-01 2010-07-01 false Bacillus pumilus GB34; exemption from... FOOD Exemptions From Tolerances § 180.1224 Bacillus pumilus GB34; exemption from the requirement of a... pesticide Bacillus pumilus GB34 when used as a seed treatment in or on all food commodities. An exemption...

  14. 40 CFR 180.1224 - Bacillus pumilus GB34; exemption from the requirement of a tolerance.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 40 Protection of Environment 25 2012-07-01 2012-07-01 false Bacillus pumilus GB34; exemption from... FOOD Exemptions From Tolerances § 180.1224 Bacillus pumilus GB34; exemption from the requirement of a... pesticide Bacillus pumilus GB34 when used as a seed treatment in or on all food commodities. An exemption...

  15. Genomic Changes that May be Responsible for the Elevated UV Resistance of Bacillus Pumilus SAFR-032

    NASA Astrophysics Data System (ADS)

    Tirumalai, M. R.; Rastogi, R.; Venkateswaran, K.; Fox, G. E.

    2010-04-01

    The genomes of Bacillus pumilus SAFR-032 whose spores are highly resistant to UV and the closely related B. pumilus ATCC-7061 that lacks this resistance are compared. Candidate genes are identified that may be responsible for the elevated resistance.

  16. Genome Sequence of Bacillus pumilus S-1, an Efficient Isoeugenol-Utilizing Producer for Natural Vanillin

    PubMed Central

    Su, Fei; Hua, Dongliang; Zhang, Zhaobin; Wang, Xiaoyu; Tang, Hongzhi; Tao, Fei; Tai, Cui; Wu, Qiulin; Wu, Geng; Xu, Ping

    2011-01-01

    Bacillus pumilus S-1 is an efficient isoeugenol-utilizing producer of natural vanillin. The genome of B. pumilus S-1 contains the epoxide hydrolase and six candidate monooxygenases that make it possible to explore the mechanism involved in conversion of isoenguenol to vanillin in the B. pumilus strain. PMID:22038964

  17. 40 CFR 180.1255 - Bacillus pumilus strain QST 2808; exemption from the requirement of a tolerance.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 23 2010-07-01 2010-07-01 false Bacillus pumilus strain QST 2808... RESIDUES IN FOOD Exemptions From Tolerances § 180.1255 Bacillus pumilus strain QST 2808; exemption from the... the microbial pesticide Bacillus pumilus strain QST 2808 when used in or on all...

  18. 40 CFR 180.1313 - Bacillus pumilus strain GHA 180; exemption from the requirement of a tolerance.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 40 Protection of Environment 25 2012-07-01 2012-07-01 false Bacillus pumilus strain GHA 180... RESIDUES IN FOOD Exemptions From Tolerances § 180.1313 Bacillus pumilus strain GHA 180; exemption from the... Bacillus pumilus strain GHA 180 in or on all food commodities when used in accordance with...

  19. 40 CFR 180.1226 - Bacillus pumilus strain QST2808; temporary exemption from the requirement of a tolerance.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 40 Protection of Environment 24 2014-07-01 2014-07-01 false Bacillus pumilus strain QST2808... RESIDUES IN FOOD Exemptions From Tolerances § 180.1226 Bacillus pumilus strain QST2808; temporary exemption... established for residues of the microbial pesticide Bacillus pumilus strain QST2808 when used in or on...

  20. 40 CFR 180.1255 - Bacillus pumilus strain QST 2808; exemption from the requirement of a tolerance.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 40 Protection of Environment 25 2013-07-01 2013-07-01 false Bacillus pumilus strain QST 2808... RESIDUES IN FOOD Exemptions From Tolerances § 180.1255 Bacillus pumilus strain QST 2808; exemption from the... the microbial pesticide Bacillus pumilus strain QST 2808 when used in or on all...

  1. 40 CFR 180.1226 - Bacillus pumilus strain QST2808; temporary exemption from the requirement of a tolerance.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 40 Protection of Environment 25 2013-07-01 2013-07-01 false Bacillus pumilus strain QST2808... RESIDUES IN FOOD Exemptions From Tolerances § 180.1226 Bacillus pumilus strain QST2808; temporary exemption... established for residues of the microbial pesticide Bacillus pumilus strain QST2808 when used in or on...

  2. 40 CFR 180.1313 - Bacillus pumilus strain GHA 180; exemption from the requirement of a tolerance.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 40 Protection of Environment 25 2013-07-01 2013-07-01 false Bacillus pumilus strain GHA 180... RESIDUES IN FOOD Exemptions From Tolerances § 180.1313 Bacillus pumilus strain GHA 180; exemption from the... Bacillus pumilus strain GHA 180 in or on all food commodities when used in accordance with...

  3. 40 CFR 180.1255 - Bacillus pumilus strain QST 2808; exemption from the requirement of a tolerance.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 40 Protection of Environment 24 2011-07-01 2011-07-01 false Bacillus pumilus strain QST 2808... RESIDUES IN FOOD Exemptions From Tolerances § 180.1255 Bacillus pumilus strain QST 2808; exemption from the... the microbial pesticide Bacillus pumilus strain QST 2808 when used in or on all...

  4. 40 CFR 180.1226 - Bacillus pumilus strain QST2808; temporary exemption from the requirement of a tolerance.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 40 Protection of Environment 24 2011-07-01 2011-07-01 false Bacillus pumilus strain QST2808... RESIDUES IN FOOD Exemptions From Tolerances § 180.1226 Bacillus pumilus strain QST2808; temporary exemption... established for residues of the microbial pesticide Bacillus pumilus strain QST2808 when used in or on...

  5. 40 CFR 180.1255 - Bacillus pumilus strain QST 2808; exemption from the requirement of a tolerance.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 40 Protection of Environment 24 2014-07-01 2014-07-01 false Bacillus pumilus strain QST 2808... RESIDUES IN FOOD Exemptions From Tolerances § 180.1255 Bacillus pumilus strain QST 2808; exemption from the... the microbial pesticide Bacillus pumilus strain QST 2808 when used in or on all...

  6. 40 CFR 180.1226 - Bacillus pumilus strain QST2808; temporary exemption from the requirement of a tolerance.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 23 2010-07-01 2010-07-01 false Bacillus pumilus strain QST2808... RESIDUES IN FOOD Exemptions From Tolerances § 180.1226 Bacillus pumilus strain QST2808; temporary exemption... established for residues of the microbial pesticide Bacillus pumilus strain QST2808 when used in or on...

  7. 40 CFR 180.1322 - Bacillus pumilus strain BU F-33; exemption from the requirement of a tolerance.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 40 Protection of Environment 25 2013-07-01 2013-07-01 false Bacillus pumilus strain BU F-33... RESIDUES IN FOOD Exemptions From Tolerances § 180.1322 Bacillus pumilus strain BU F-33; exemption from the... Bacillus pumilus strain BU F-33 in or on all food commodities when applied to elicit induced...

  8. 40 CFR 180.1322 - Bacillus pumilus strain BU F-33; exemption from the requirement of a tolerance.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 40 Protection of Environment 24 2014-07-01 2014-07-01 false Bacillus pumilus strain BU F-33... RESIDUES IN FOOD Exemptions From Tolerances § 180.1322 Bacillus pumilus strain BU F-33; exemption from the... Bacillus pumilus strain BU F-33 in or on all food commodities when applied to elicit induced...

  9. 40 CFR 180.1255 - Bacillus pumilus strain QST 2808; exemption from the requirement of a tolerance.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 40 Protection of Environment 25 2012-07-01 2012-07-01 false Bacillus pumilus strain QST 2808... RESIDUES IN FOOD Exemptions From Tolerances § 180.1255 Bacillus pumilus strain QST 2808; exemption from the... the microbial pesticide Bacillus pumilus strain QST 2808 when used in or on all...

  10. 40 CFR 180.1313 - Bacillus pumilus strain GHA 180; exemption from the requirement of a tolerance.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 40 Protection of Environment 24 2014-07-01 2014-07-01 false Bacillus pumilus strain GHA 180... RESIDUES IN FOOD Exemptions From Tolerances § 180.1313 Bacillus pumilus strain GHA 180; exemption from the... Bacillus pumilus strain GHA 180 in or on all food commodities when used in accordance with...

  11. 40 CFR 180.1226 - Bacillus pumilus strain QST2808; temporary exemption from the requirement of a tolerance.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 40 Protection of Environment 25 2012-07-01 2012-07-01 false Bacillus pumilus strain QST2808... RESIDUES IN FOOD Exemptions From Tolerances § 180.1226 Bacillus pumilus strain QST2808; temporary exemption... established for residues of the microbial pesticide Bacillus pumilus strain QST2808 when used in or on...

  12. Salt stress represses production of extracellular proteases in Bacillus pumilus.

    PubMed

    Liu, R F; Huang, C L; Feng, H

    2015-01-01

    Bacillus pumilus is able to secrete subtilisin-like prote-ases, one of which has been purified and characterized biochemically, demonstrating great potential for use in industrial applications. In the current study, the biosynthesis and transcription of extracellular pro-teases in B. pumilus (BA06) under salt stress were investigated using various methods, including a proteolytic assay, zymogram analysis, and real-time PCR. Our results showed that total extracellular proteolytic activity, both in fermentation broth and on milk-containing agar plates, was considerably repressed by salt in a dosage-dependent manner. As Bacillus species usually secret multiple extracellular proteases, a vari-ety of individual extracellular protease encoding genes were selected for real-time PCR analysis. It was shown that proteases encoded by the aprE and aprX genes were the major proteases in the fermentation broth in terms of their transcripts in B. pumilus. Further, transcription of aprE, aprX, and epr genes was indeed repressed by salt stress. In con-trast, transcription of other genes (e.g., vpr and wprA) was not repressed or significantly affected by the salt. Conclusively, salt stress represses total extracellular proteolytic activity in B. pumilus, which can largely be ascribed to suppression of the major protease-encoding genes (aprE, aprX) at the transcriptional level. In contrast, transcription of other pro-tease-encoding genes (e.g., vpr, wprA) was not repressed by salt stress. PMID:25966269

  13. [Expression optimization of Bacillus pumilus subtilisin-like proteinase].

    PubMed

    Cheremin, A M; Niamsuren, Ch; To?mentseva, A A; Sharipova, M R

    2014-01-01

    The characteristics of Bacillus pumilus subtilisin-like protease expression were investigated. Potential binding sites for the transcription factor DegU~P have been identified in the regulatory region of the enzyme gene, which have been optimized for interaction with the regulatory protein. The expression of the extracellular subtilisin-like protease modified gene has been examined. It was established that the modification of one of the sites has resulted in increased expression of the proteinase in 2 times. It is concluded that the optimization of the promoter led to increased expression of subtilisin-like protease. PMID:25895372

  14. Evaluation of Bacillus cereus and Bacillus pumilus metabolites for anthelmintic activity

    PubMed Central

    Kumar, M. L. Vijaya; Thippeswamy, B.; Kuppust, I. L.; Naveenkumar, K. J.; Shivakumar, C. K.

    2015-01-01

    Objective: To assess the anthelmintic acivity of Bacillus cereus and Bacillus pumilus metabolites. Materials and Methods: The successive solvent extractions with petroleum ether, ethyl acetate and methanol. The solvent extracts were tested for anthelmintic activity against Pheretima posthuma at 20 mg/ml concentration. The time of paralysis and time of death of the worms was determined for all the extracts. Albendazole was taken as a standard reference and sterile water as a control. Results: All the sample extracts showed significant anthelmintic activity in paralyzing the worms comparable with that of the standard drug. The time of death exhibited by BP metabolites was close to the time exhibited by standard. Conclusion: The study indicates both bacteria Bacillus cereus and Bacillus pumilus have anthelmintic activity indicating potential metabolites in them. PMID:25598639

  15. Genetic Analysis in Bacillus pumilus by PBS1-Mediated Transduction

    PubMed Central

    Lovett, Paul S.; Young, Frank E.

    1970-01-01

    Bacteriophage PBS1 mediates generalized transduction in Bacillus pumilus NRRL B-3275 (BpB1). Transduction frequencies for single auxotrophic markers are of the order of 10−4 transductants per plaque-forming unit in crude phage lysates. The characteristics of PBS1 propagated on BpB1 and the properties of the system of transduction are similar to those reported for PBS1 propagated on Bacillus subtilis. By transduction, eight amino acid auxotrophic markers in BpB1 have been oriented into two linkage groups. One group contains the auxotrophic markers arginine A, leucine, and phenylalanine, and the other group contains the markers lysine, serine, tryptophan, isoleucine-valine, and isoleucine. The nature and relative order of the markers within each linkage group suggest that the arrangement of genes in these areas of the chromosome of BpB1 is similar to the arrangement of phenotypically comparable genes in two linkage groups (defined by PBS1 transduction) in B. subtilis. However, transduction of any of the above cited markers in BpB1 to prototrophy with PBS1 propagated on B. subtilis 168 could not be demonstrated. In addition to BpB1, seven other strains of B. pumilus can be infected with PBS1. Transduction has been demonstrated in three of these strains. Images PMID:5413829

  16. Draft Genome Sequence of the Shellfish Larval Probiotic Bacillus pumilus RI06-95

    PubMed Central

    Hamblin, Meagan; Spinard, Edward; Gomez-Chiarri, Marta; Nelson, David R.

    2015-01-01

    Bacillus pumilus RI06-95 is a marine bacterium isolated in Narragansett, Rhode Island, which has shown probiotic activity against marine pathogens in larval shellfish. We report the genome of B. pumilus RI06-95, which provides insight into the microbe’s probiotic ability and may be used in future studies of the probiotic mechanism. PMID:26337873

  17. Draft Genome Sequence of the Shellfish Larval Probiotic Bacillus pumilus RI06-95.

    PubMed

    Hamblin, Meagan; Spinard, Edward; Gomez-Chiarri, Marta; Nelson, David R; Rowley, David C

    2015-01-01

    Bacillus pumilus RI06-95 is a marine bacterium isolated in Narragansett, Rhode Island, which has shown probiotic activity against marine pathogens in larval shellfish. We report the genome of B. pumilus RI06-95, which provides insight into the microbe's probiotic ability and may be used in future studies of the probiotic mechanism. PMID:26337873

  18. Identification of Bacillus subtilis NRRL B-3275 as a strain of Bacillus pumilus.

    PubMed

    Lovett, P S; Young, F E

    1969-11-01

    The physiological and biochemical properties of a species of Bacillus previously identified as B. subtilis NRRL B-3275 (B-3275) were compared with those of seven strains of B. pumilus and five strains of B. subtilis. The biotin requirement of B-3275, its inability to hydrolyze starch, and its failure to reduce nitrate indicate that the organism is more closely related to the B. pumilus strains than to those of B. subtilis. Hybridization of deoxyribonucleic acid (DNA) from B-3275 with that of the strains of B. pumilus showed a binding efficiency (compared with the homologous reaction) of 58 to 99%, depending on the strain. Hybridization with the DNA from any of the strains of B. subtilis did not exceed 24%. DNA from B-3275 was unable to transform two amino acid auxotrophic markers to prototrophy in a highly competent strain of B. subtilis 168. We conclude that B-3275 is a strain of B. pumilus which we designate as B. pumilus NRRL B-3275. PMID:4982197

  19. Complete Genome Sequence of Bacillus pumilus Strain WP8, an Efficient Plant Growth-Promoting Rhizobacterium.

    PubMed

    Kang, Yijun; Shen, Min; Wang, Huanli; Zhao, Qingxin

    2015-01-01

    Bacillus pumilus strain WP8 is an efficient plant growth-promoting rhizobacterium. Here, we present the complete genome of WP8 and its genes involved in plant growth promotion and biocontrol. PMID:25614565

  20. Antitrypanosomal alkaloids from the marine bacterium Bacillus pumilus.

    PubMed

    Martínez-Luis, Sergio; Gómez, José Félix; Spadafora, Carmenza; Guzmán, Héctor M; Gutiérrez, Marcelino

    2012-01-01

    Fractionation of the ethyl acetate extract of the marine bacterium Bacillus pumilus isolated from the black coral Antipathes sp. led to the isolation of five compounds: cyclo-(L-Leu-L-Pro) (1), 3-hydroxyacetylindole (2), N-acetyl-?-oxotryptamine (3), cyclo-(L-Phe-L-Pro) (4), and 3-formylindole (5). The structures of compounds 1-5 were established by spectroscopic analyses, including HRESITOF-MS and NMR (1H, 13C, HSQC, HMBC and COSY). Compounds 2, 3 and 5 caused the inhibition on the growth of Trypanosoma cruzi (T. cruzi), with IC50 values of 20.6, 19.4 and 26.9 ?M, respectively, with moderate cytotoxicity against Vero cells. Compounds 1-5 were found to be inactive when tested against Plasmodium falciparum and Leishmania donovani, therefore showing selectivity against T. cruzi parasites. PMID:22990456

  1. Construction of novel shuttle expression vectors for gene expression in Bacillus subtilis and Bacillus pumilus.

    PubMed

    Shao, Huanhuan; Cao, Qinghua; Zhao, Hongyan; Tan, Xuemei; Feng, Hong

    2015-01-01

    A native plasmid (pSU01) was detected by genome sequencing of Bacillus subtilis strain S1-4. Two pSU01-based shuttle expression vectors pSU02-AP and pSU03-AP were constructed enabling stable replication in B. subtilis WB600. These vectors contained the reporter gene aprE, encoding an alkaline protease from Bacillus pumilus BA06. The expression vector pSU03-AP only possessed the minimal replication elements (rep, SSO, DSO) and exhibited more stability on structure, suggesting that the rest of the genes in pSU01 (ORF1, ORF2, mob, hsp) were unessential for the structural stability of plasmid in B. subtilis. In addition, recombinant production of the alkaline protease was achieved more efficiently with pSU03-AP whose copy number was estimated to be more than 100 per chromosome. Furthermore, pSU03-AP could also be used to transform and replicate in B. pumilus BA06 under selective pressure. In conclusion, pSU03-AP is expected to be a useful tool for gene expression in Bacillus subtilis and B. pumilus. PMID:26377132

  2. Microbiological bioassay using Bacillus pumilus to detect tetracycline in milk.

    PubMed

    Tumini, Melisa; Nagel, Orlando Guillermo; Althaus, Rafael Lisandro

    2015-05-01

    The tetracyclines (TCs) are widely used in the treatment of several diseases of cattle and their residues may be present in milk. To control these residues it is necessary to have available inexpensive screening methods, user-friendly and capable of analysing a high number of samples. The purpose of this study was to design a bioassay of microbiological inhibition in microtiter plates with spores of Bacillus pumilus to detect TCs at concentrations corresponding to the Maximum Residue Limits (MRLs). Several complementary experiments were performed to design the bioassay. In the first study, we determined the concentration of spores that produce a change in the bioassay's relative absorbance in a short time period. Subsequently, we assessed the concentration of chloramphenicol required to decrease the detection limit (DL) of TCs at MRLs levels. Thereafter, specificity, DL and cross-specificity of the bioassay were estimated. The most appropriate microbiological inhibition assay had a B. pumilus concentration of 1.6 × 10(9) spores/ml, fortified with 2500 ?g chloramphenicol/l (CAP) in Mueller Hinton culture medium using brilliant black and toluidine blue as redox indicator. This bioassay detected 117 ?g chlortetracycline/l, 142 ?g oxytetracycline/l and 105 ?g tetracycline/l by means of a change in the indicator's colour in a period of 5 h. The method showed good specificity (97.9%) which decreased slightly (93.3%) in milk samples with high somatic cell counts (>250,000 cells/ml). Furthermore, other antimicrobials studied (except neomycin) must be present in milk at high concentrations (from >5 to >100 MRLs) to produce positive results in this assay, indicating a low cross specificity. PMID:25720802

  3. Study of the radiation effect of (99)Mo/(99m)Tc generator on Bacillus subtilis and Bacillus pumilus species.

    PubMed

    Fukumori, Neuza T O; Endo, Erica M M; Felgueiras, Carlos F; Matsuda, Margareth M N; Osso Junior, João A

    2016-01-01

    In this work, molybdenum-99 loaded columns were challenged with Bacillus subtilis vegetative cells and Bacillus pumilus spores inside and outside the alumina column, and microbial recovery and radiation effect were assessed. Alumina was a barrier for the passage of microorganisms regardless the species, whilst spores were more retained than vegetative cells with a lower microbial recovery, without significant differences between 9.25 and 74GBq generators. Bacillus pumilus biological indicator showed lower recoveries, suggesting a radiation inactivating effect on microorganisms. PMID:26408912

  4. Biochemical and molecular characterization of Bacillus pumilus isolated from coastal environment in Cochin, India

    PubMed Central

    Parvathi, Ammini; Krishna, Kiran; Jose, Jiya; Joseph, Neetha; Nair, Santha

    2009-01-01

    Bacillus species constitute a diverse group of bacteria widely distributed in soil and the aquatic environment. In this study, Bacillus strains isolated from the coastal environment of Cochin, India were identified by detailed conventional biochemical methods, fatty acid methyl ester (FAME) analysis and partial 16S rDNA sequencing. Analysis of the data revealed that Bacillus pumilus was the most predominant species in the region under study followed by B. cereus and B. sphaericus. The B. pumilus isolates were further characterized by arbitrarily primed PCR (AP-PCR), antibiotic sensitivity profiling and PCR screening for known toxin genes associated with Bacillus spp. All B. pumilus isolates were biochemically identical, exhibited high protease and lipase activity and uniformly sensitive to antibiotics tested in this study. One strain of B. pumilus harboured cereulide synthetase gene cesB of B. cereus which was indistinguishable from rest of the isolates biochemically and by AP-PCR. This study reports, for the first time, the presence of the emetic toxin gene cesB in B. pumilus. PMID:24031357

  5. Molecular detection of a bacterial contaminant Bacillus pumilus in symptomless potato plant tissue cultures.

    PubMed

    Isenegger, D A; Taylor, P W J; Mullins, K; McGregor, G R; Barlass, M; Hutchinson, J F

    2003-04-01

    An aberrant random amplified polymorphic DNA (RAPD) marker in genomic DNA of tissue culture plantlets was frequently observed during a comparison of DNA fingerprints derived from potato germplasm grown in tissue culture and the field. The RAPD marker was cloned, sequenced and determined to be of bacterial origin. A bacterial contaminant was isolated from the tissue culture plants and identified as a Bacillus pumilus. A set of sequence characterised amplified region (SCAR) primers were designed from the sequence of the cloned fragment and tested for the specific detection of B. pumilus. Polymerase chain reaction-restriction fragment length polymorphisms (PCR-RFLPs) were also used to generate B. pumilus profiles specific to our isolate in order to test and confirm the sequence homology of amplified markers generated from a range of DNA samples isolated from tissue culture plants and pure isolates of B. pumilus-like bacteria. PMID:12789528

  6. Differentiation of Bacillus pumilus and Bacillus safensis using MALDI-TOF-MS.

    PubMed

    Branquinho, Raquel; Sousa, Clara; Lopes, João; Pintado, Manuela E; Peixe, Luísa V; Osório, Hugo

    2014-01-01

    Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) despite being increasingly used as a method for microbial identification, still present limitations in which concerns the differentiation of closely related species. Bacillus pumillus and Bacillus safensis, are species of biotechnological and pharmaceutical significance, difficult to differentiate by conventional methodologies. In this study, using a well-characterized collection of B. pumillus and B. safensis isolates, we demonstrated the suitability of MALDI-TOF-MS combined with chemometrics to accurately and rapidly identify them. Moreover, characteristic species-specific ion masses were tentatively assigned, using UniProtKB/Swiss-Prot and UniProtKB/TrEMBL databases and primary literature. Delineation of B. pumilus (ions at m/z 5271 and 6122) and B. safensis (ions at m/z 5288, 5568 and 6413) species were supported by a congruent characteristic protein pattern. Moreover, using a chemometric approach, the score plot created by partial least square discriminant analysis (PLSDA) of mass spectra demonstrated the presence of two individualized clusters, each one enclosing isolates belonging to a species-specific spectral group. The generated pool of species-specific proteins comprised mostly ribosomal and SASPs proteins. Therefore, in B. pumilus the specific ion at m/z 5271 was associated with a small acid-soluble spore protein (SASP O) or with 50S protein L35, whereas in B. safensis specific ions at m/z 5288 and 5568 were associated with SASP J and P, respectively, and an ion at m/z 6413 with 50S protein L32. Thus, the resulting unique protein profile combined with chemometric analysis, proved to be valuable tools for B. pumilus and B. safensis discrimination, allowing their reliable, reproducible and rapid identification. PMID:25314655

  7. Differentiation of Bacillus pumilus and Bacillus safensis Using MALDI-TOF-MS

    PubMed Central

    Branquinho, Raquel; Sousa, Clara; Lopes, João; Pintado, Manuela E.; Peixe, Luísa V.; Osório, Hugo

    2014-01-01

    Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) despite being increasingly used as a method for microbial identification, still present limitations in which concerns the differentiation of closely related species. Bacillus pumillus and Bacillus safensis, are species of biotechnological and pharmaceutical significance, difficult to differentiate by conventional methodologies. In this study, using a well-characterized collection of B. pumillus and B. safensis isolates, we demonstrated the suitability of MALDI-TOF-MS combined with chemometrics to accurately and rapidly identify them. Moreover, characteristic species-specific ion masses were tentatively assigned, using UniProtKB/Swiss-Prot and UniProtKB/TrEMBL databases and primary literature. Delineation of B. pumilus (ions at m/z 5271 and 6122) and B. safensis (ions at m/z 5288, 5568 and 6413) species were supported by a congruent characteristic protein pattern. Moreover, using a chemometric approach, the score plot created by partial least square discriminant analysis (PLSDA) of mass spectra demonstrated the presence of two individualized clusters, each one enclosing isolates belonging to a species-specific spectral group. The generated pool of species-specific proteins comprised mostly ribosomal and SASPs proteins. Therefore, in B. pumilus the specific ion at m/z 5271 was associated with a small acid-soluble spore protein (SASP O) or with 50S protein L35, whereas in B. safensis specific ions at m/z 5288 and 5568 were associated with SASP J and P, respectively, and an ion at m/z 6413 with 50S protein L32. Thus, the resulting unique protein profile combined with chemometric analysis, proved to be valuable tools for B. pumilus and B. safensis discrimination, allowing their reliable, reproducible and rapid identification. PMID:25314655

  8. Recurrent isolation of hydrogen peroxide-resistant spores of Bacillus pumilus from a spacecraft assembly facility

    NASA Technical Reports Server (NTRS)

    Kempf, Michael J.; Chen, Fei; Kern, Roger; Venkateswaran, Kasthuri

    2005-01-01

    While the microbial diversity of a spacecraft assembly facility at the Jet Propulsion Laboratory (Pasadena, CA) was being monitored, H2O2-resistant bacterial strains were repeatedly isolated from various surface locations. H2O2 is a possible sterilant for spacecraft hardware because it is a low-temperature process and compatible with various modern-day spacecraft materials, electronics, and components. Both conventional biochemical testing and molecular analyses identified these strains as Bacillus pumilus. This Bacillus species was found in both unclassified (entrance floors, anteroom, and air-lock) and classified (floors, cabinet tops, and air) locations. Both vegetative cells and spores of several B. pumilus isolates were exposed to 5% liquid H2O2 for 60 min. Spores of each strain exhibited higher resistance than their respective vegetative cells to liquid H2O2. Results indicate that the H2O2 resistance observed in both vegetative cells and spores is strain-specific, as certain B. pumilus strains were two to three times more resistant than a standard Bacillus subtilis dosimetry strain. An example of this trend was observed when the type strain of B. pumilus, ATCC 7061, proved sensitive, whereas several environmental strains exhibited varying degrees of resistance, to H2O2. Repeated isolation of H2O2-resistant strains of B. pumilus in a clean-room is a concern because their persistence might potentially compromise life-detection missions, which have very strict cleanliness and sterility requirements for spacecraft hardware.

  9. Recurrent Isolation of Hydrogen Peroxide-Resistant Spores of Bacillus pumilus from a Spacecraft Assembly Facility

    NASA Astrophysics Data System (ADS)

    Kempf, Michael J.; Chen, Fei; Kern, Roger; Venkateswaran, Kasthuri

    2005-06-01

    While the microbial diversity of a spacecraft assembly facility at the Jet Propulsion Laboratory (Pasadena, CA) was being monitored, H2O2-resistant bacterial strains were repeatedly isolated from various surface locations. H2O2 is a possible sterilant for spacecraft hardware because it is a low-temperature process and compatible with various modern-day spacecraft materials, electronics, and components. Both conventional biochemical testing and molecular analyses identified these strains as Bacillus pumilus. This Bacillus species was found in both unclassified (entrance floors, anteroom, and air-lock) and classified (floors, cabinet tops, and air) locations. Both vegetative cells and spores of several B. pumilus isolates were exposed to 5% liquid H2O2 for 60 min. Spores of each strain exhibited higher resistance than their respective vegetative cells to liquid H2O2. Results indicate that the H2O2 resistance observed in both vegetative cells and spores is strain-specific, as certain B. pumilus strains were two to three times more resistant than a standard Bacillus subtilis dosimetry strain. An example of this trend was observed when the type strain of B. pumilus, ATCC 7061, proved sensitive, whereas several environmental strains exhibited varying degrees of resistance, to H2O2. Repeated isolation of H2O2-resistant strains of B. pumilus in a clean-room is a concern because their persistence might potentially compromise life-detection missions, which have very strict cleanliness and sterility requirements for spacecraft hardware.

  10. [Cloning, characterization and application of the promoter of alkaline protease gene in Bacillus pumilus].

    PubMed

    Yang, Chun-Hui; Wang, Hai-Yan

    2007-07-01

    A 797 bp promoter fragment of alkaline protease gene was cloned from the Bacillus pumilus genome by employing TAIL-PCR strategy. The sequence analysis of this promoter fragment showed that the sequence accounting for gene expression was approximately 390bp. Deletion analysis of the fragment defined the minimal required sequence of promoter for initiating transcription lies on a 160 bp region upstream of the start codon. The alkaline protease gene WApQ3 containing the cloned promoter fragment was inserted into the shuttle vector pSUGV4 and the constructed expression plasmid pSUBpWApQ3 was transformed into Bacillus subtilis and B. pumilus. Active alkaline protease was successfully expressed in both host strains. The peak value of extracellular alkaline protease activities of B. subtilis and B. pumilus recombinants reached to 465.5 U/mL and 3060 U/mL, respectively. PMID:17646155

  11. Bacillus pumilus Reveals a Remarkably High Resistance to Hydrogen Peroxide Provoked Oxidative Stress

    PubMed Central

    Jürgen, Britta; Methling, Karen; Schlüter, Rabea; Albrecht, Dirk; van Hijum, Sacha A. F. T.; Bongaerts, Johannes; Maurer, Karl-Heinz; Lalk, Michael; Schweder, Thomas; Hecker, Michael; Voigt, Birgit

    2014-01-01

    Bacillus pumilus is characterized by a higher oxidative stress resistance than other comparable industrially relevant Bacilli such as B. subtilis or B. licheniformis. In this study the response of B. pumilus to oxidative stress was investigated during a treatment with high concentrations of hydrogen peroxide at the proteome, transcriptome and metabolome level. Genes/proteins belonging to regulons, which are known to have important functions in the oxidative stress response of other organisms, were found to be upregulated, such as the Fur, Spx, SOS or CtsR regulon. Strikingly, parts of the fundamental PerR regulon responding to peroxide stress in B. subtilis are not encoded in the B. pumilus genome. Thus, B. pumilus misses the catalase KatA, the DNA-protection protein MrgA or the alkyl hydroperoxide reductase AhpCF. Data of this study suggests that the catalase KatX2 takes over the function of the missing KatA in the oxidative stress response of B. pumilus. The genome-wide expression analysis revealed an induction of bacillithiol (Cys-GlcN-malate, BSH) relevant genes. An analysis of the intracellular metabolites detected high intracellular levels of this protective metabolite, which indicates the importance of bacillithiol in the peroxide stress resistance of B. pumilus. PMID:24465625

  12. Bacillus pumilus reveals a remarkably high resistance to hydrogen peroxide provoked oxidative stress.

    PubMed

    Handtke, Stefan; Schroeter, Rebecca; Jürgen, Britta; Methling, Karen; Schlüter, Rabea; Albrecht, Dirk; van Hijum, Sacha A F T; Bongaerts, Johannes; Maurer, Karl-Heinz; Lalk, Michael; Schweder, Thomas; Hecker, Michael; Voigt, Birgit

    2014-01-01

    Bacillus pumilus is characterized by a higher oxidative stress resistance than other comparable industrially relevant Bacilli such as B. subtilis or B. licheniformis. In this study the response of B. pumilus to oxidative stress was investigated during a treatment with high concentrations of hydrogen peroxide at the proteome, transcriptome and metabolome level. Genes/proteins belonging to regulons, which are known to have important functions in the oxidative stress response of other organisms, were found to be upregulated, such as the Fur, Spx, SOS or CtsR regulon. Strikingly, parts of the fundamental PerR regulon responding to peroxide stress in B. subtilis are not encoded in the B. pumilus genome. Thus, B. pumilus misses the catalase KatA, the DNA-protection protein MrgA or the alkyl hydroperoxide reductase AhpCF. Data of this study suggests that the catalase KatX2 takes over the function of the missing KatA in the oxidative stress response of B. pumilus. The genome-wide expression analysis revealed an induction of bacillithiol (Cys-GlcN-malate, BSH) relevant genes. An analysis of the intracellular metabolites detected high intracellular levels of this protective metabolite, which indicates the importance of bacillithiol in the peroxide stress resistance of B. pumilus. PMID:24465625

  13. Cell physiology of the biotechnological relevant bacterium Bacillus pumilus-an omics-based approach.

    PubMed

    Handtke, Stefan; Volland, Sonja; Methling, Karen; Albrecht, Dirk; Becher, Dörte; Nehls, Jenny; Bongaerts, Johannes; Maurer, Karl-Heinz; Lalk, Michael; Liesegang, Heiko; Voigt, Birgit; Daniel, Rolf; Hecker, Michael

    2014-12-20

    Members of the species Bacillus pumilus get more and more in focus of the biotechnological industry as potential new production strains. Based on exoproteome analysis, B. pumilus strain Jo2, possessing a high secretion capability, was chosen for an omics-based investigation. The proteome and metabolome of B. pumilus cells growing either in minimal or complex medium was analyzed. In total, 1542 proteins were identified in growing B. pumilus cells, among them 1182 cytosolic proteins, 297 membrane and lipoproteins and 63 secreted proteins. This accounts for about 43% of the 3616 proteins encoded in the B. pumilus Jo2 genome sequence. By using GC-MS, IP-LC/MS and H NMR methods numerous metabolites were analyzed and assigned to reconstructed metabolic pathways. In the genome sequence a functional secretion system including the components of the Sec- and Tat-secretion machinery was found. Analysis of the exoproteome revealed secretion of about 70 proteins with predicted secretion signals. In addition, selected production-relevant genome features such as restriction modification systems and NRPS clusters of B. pumilus Jo2 are discussed. PMID:25281541

  14. Molecular analysis of endophytic bacteria from the genus Bacillus isolated from tropical maize (Zea mays L.)

    PubMed Central

    Figueiredo, José Edson Fontes; Gomes, Eliane Aparecida; Guimarães, Claudia Teixeira; de Paula Lana, Ubiraci Gomes; Teixeira, Marta Aparecida; Lima, Guilherme Vitor Corrêa; Bressan, Wellington

    2009-01-01

    Endophytic bacteria play an important role in agriculture by improving plant performance and adaptation against biotic and abiotic stresses. In the present study molecular methods were used for identifying Bacillus endophytic bacteria isolated from Brazilian sweet corn. SDS-PAGE of whole-cell protein extract of forty-two isolates revealed a high number of scrutinable bands. Twenty-four isolates were identified in nine different groups of duplicated bacteria and eighteen were identified as unique. Some high-accumulated polipeptides with variable length were observed in almost isolates. Partial sequencing of 16S ribosomal gene revealed that all isolates are Bacillus sp. and among thirteen isolates with similar protein profiles, two were different strains. Among the forty-two isolates identified by rDNA sequencing, Bacillus subitilis and B. pumilus were the most frequenty species (15 and 12 isolates, respectively) followed by B. licheniformes (7 isolates), B. cereus (5 isolates) and B. amiloliquefascens (3 isolates). According to present results, SDS-PAGE technique could be used as a fast and cheap first tool for identifying inter-specific variation in maize endophytic bacterial collections while rDNA sequencing could be applied for analyzing intra-specific variation among isolates with similar protein profile as well as for taxonomic studies. PMID:24031395

  15. Genome Sequence of Bacillus pumilus Strain Bonn, Isolated from an Anthrax-Like Necrotic Skin Infection Site of a Child.

    PubMed

    Grass, Gregor; Bierbaum, Gabriele; Molitor, Ernst; Götte, Natascha; Antwerpen, Markus

    2016-01-01

    We report the draft genome sequence of Bacillus pumilus strain Bonn associated with human skin infection. B. pumilus Bonn was isolated from a carbuncle-like necrotic site, resembling cutaneous anthrax, on the back of the hand of a 10-year-old child. PMID:26868410

  16. Genome Sequence of Bacillus pumilus Strain Bonn, Isolated from an Anthrax-Like Necrotic Skin Infection Site of a Child

    PubMed Central

    Grass, Gregor; Bierbaum, Gabriele; Molitor, Ernst; Götte, Natascha

    2016-01-01

    We report the draft genome sequence of Bacillus pumilus strain Bonn associated with human skin infection. B. pumilus Bonn was isolated from a carbuncle-like necrotic site, resembling cutaneous anthrax, on the back of the hand of a 10-year-old child. PMID:26868410

  17. Cloning of high activity xylanase gene from Bacillus pumilus PJ19.

    PubMed

    Hamzah, Ainon; Abdulrashid, Nooraini

    2002-10-01

    The xylanase gene from Bacillus pumilus PJ19 amplified by polymerase chain reaction (PCR) was cloned into pCRII vector and transformed into Escherichia coli strain INValphaF'. Starting from an ATG as an initiator codon, an open reading frame coding for 202 amino acids was obtained. The recombinant xylanase sequence showed a 96% homology with the xylanase sequence from B. pumilus IPO strain and had an estimated molecular weight of 22,474. Xylanase activity expressed by E. coli INValphaF' harboring the cloned gene was located primarily in the cytoplasmic fraction. PMID:12385974

  18. Modification of a xylanase from bacillus pumilus with pentaammineruthenium(III)

    SciTech Connect

    Evans, B.R.; Lane, L.; Woodward, J.

    1996-10-01

    A xylanase, xynA of Bacillus pumilus, was purified, and then modified by the attachment of pentaammineruthenium, resulting in the generation of a xylanase with veratryl alcohol oxidase activity. Modification of B. pumilus xyn A was found to greatly reduce xylan hydrolysis unless the active site of the xylanase was protected with xylose during the modification. Addition of histidine, cysteine, or reduced glutathione during xylan hydrolysis greatly increased xylanase activity of the modified xylanase. Glycine, glutamic acid, methionine, or oxidized glutathione had no effect on xylanase activity. The site of attachment of pentaammineruthenium, was identified as His-160 by mass spectroscopy and sequence determination of tryptic peptides from modified and native xylanase.

  19. Antibiotic production and biocontrol activity by Bacillus subtilis CL27 and Bacillus pumilus CL45.

    PubMed

    Leifert, C; Li, H; Chidburee, S; Hampson, S; Workman, S; Sigee, D; Epton, H A; Harbour, A

    1995-02-01

    Bacillus subtilis CL27 and B. pumilus CL45 showed similar activity against Botrytis cinerea in in vitro plate assays. In a seedling bioassay, however, B. subtilis CL27 had activity similar to a commercial fungicide while B. pumilus CL45 failed completely to prevent seedling damping-off caused by Bot. cinerea. Antibiotic production by the two Bacillus strains was found to depend on the growth substrate and highest antibiotic production was found on media based on homogenized cabbage tissue. Antibiotic activity was found to depend on the pH and nutrient concentration in the assay medium. Antifungal antibiotics produced by B. subtilis CL27 and B. pumilus CL45 in different fermentation media were separated by thin layer chromatography. As suspected from the activity spectrum, three antibiotics (one with activity against Alternaria brassicicola, one with activity against Botrytis cinerea and one with activity against both fungi) could be detected in the fermentation broth of CL27, but only one in the fermentation broth of CL45. The two antibiotics produced by strain CL27 with activity against A. brassicicola were identified as peptides since their bands on the TLC plates developed a green to blue/green colour after treatment with 4,4'-tetramethyldiamino-diphenylmethane (TDM) reagent. The third antibiotics produced by strain CL27 and antibiotic produced by CL45 had a similar Rf-value and appeared not to be peptides based on the reaction with TDM. However, they showed a slightly different activity spectrum when tested against a range of different fungi. Antibiotic production was clearly indicated as the mode of action of in vivo biocontrol by strain CL27 against damping off caused by Bot. cinerea of Astilbe micro-plants, because a u.v.-induced antibiotic negative mutant strain CL27b showed no activity in seedling bioassays in vivo. Also the mutant strain CL27a which produced the two peptide antibiotics but had lost the ability to produce the non-peptide antibiotic, showed greatly reduced in vivo activity. PMID:7698955

  20. Resistance and recovery studies on ultraviolet-irradiated spores of Bacillus pumilus

    SciTech Connect

    Abshire, R.L.; Bain, B.; Williams, T.

    1980-04-01

    A spore suspension model and a procedure for recovering ultraviolet (uv)-irradiated spores of Bacillus pumilus were investigated. A most-probable-number tube dilution method using double-strength Trypticase soy broth was found to be superior to the agar plate method for recovering optimal numbers of spores irradiated with sublethal doses of uv energy. Aqueous suspensions of B. pumilus survived uv doses up to 108,000 ergs/mm/sup 2/ as determined by a most-probable-number recovery and estimation procedure. Resistance and stability data were consistent and reproducible, indicating the dependability of this method for recovering uv-damaged spores. The procedures used to collect information concerning resistance characteristics for two strains of B. pumilus are discussed.

  1. 2,5-Diketopiperazines produced by Bacillus pumilus during bacteriolysis of Arthrobacter citreus.

    PubMed

    Brack, Christiane; Mikolasch, Annett; Schauer, Frieder

    2014-08-01

    We report the detection by gas chromatography/mass spectrometry and liquid chromatography/mass spectrometry analyses of the secreted 2,5-diketopiperazines (DKPs) cyclo(-Ala-Pro), cyclo(-Gly-Pro), cyclo(-Val-Pro), cyclo(-Ile-Pro), cyclo(-Leu-Pro), cyclo(-Pro-Pro), cyclo(-HyP-Pro), cyclo(-Met-Pro), and cyclo(-Phe-Pro) produced by Bacillus pumilus. The study focuses on a marine isolate and a laboratory test strain of B. pumilus with capabilities to lyse pregrown living cell lawns of different bacterial species, among them Arthrobacter citreus. Chromatographic methods were used to analyze induced bioactive compounds. At least 13 different DKPs are produced by B. pumilus. Both strains respond with an increased production of the DKPs cyclo(-Gly-Pro), cyclo(-Ala-Pro), and cyclo(-Val-Pro) to the presence of pasteurized A. citreus cells after 4 h in a nutrient-poor liquid medium. In agar diffusion assays, these DKPs did not cause lysis zones in living cell lawns, but they did inhibit further growth of several pregrown test bacteria in microplates even at concentrations as low as 1 ?g ml(-1). Antibiotic substances produced by B. pumilus after 20 h of cultivation in a special lysis medium showed lytic activity in cell-free extracts of B. pumilus culture supernatants. PMID:24449388

  2. Soybean-milk-coagulating activity of Bacillus pumilus derives from a serine proteinase.

    PubMed

    Aoyama, M; Yasuda, M; Nakachi, K; Kobamoto, N; Oku, H; Kato, F

    2000-04-01

    A proteolytic enzyme from Bacillus pumilus strain TYO-67, which was able to coagulate the protein in soybean milk, was characterized enzymologically. The optimum pH and temperature for its activities were 9.0 and 50 degrees C, respectively. The enzyme was strongly believed to be a serine proteinase because it was completely inhibited by the addition of diisopropyl fluorophosphate or phenylmethanesulfonyl fluoride. Hammerstein milk casein, cytochrome c and soybean protein were good substrates for the enzyme. Seven cleavages were detected using the oxidized insulin B-chain as peptide substrate for the proteolytic specificity test of the serine proteinase from B. pumilus. The bonds most susceptible to the action of the serine proteinase from B. pumilus were Leu-15-Tyr-16. The mode of action on soybean milk protein by the enzyme from B. pumilus was also investigated. The acidic subunit in glycinin and the alpha-, alpha- and beta-subunits in beta-conglycinin were degraded during the enzyme reaction. However, the basic subunit in glycinin could not be degraded by the enzyme. The formation of coagula in soybean milk caused by the serine proteinase from B. pumilus was mainly due to the hydrophobic interaction. PMID:10803893

  3. Transformation of Bacillus mojavensis with GFP and its endophytic localization

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Current studies on the use of bacterial endophytes for disease control and mycotoxin reduction in maize require the in planta localization and interaction of the intended biocontrol agent. Bacillus mojavensis, an endophytic species that has a phenotypic similarity to B. subtilis, has the potential...

  4. Molecular characterization of a proteolysis-resistant lipase from Bacillus pumilus SG2

    PubMed Central

    Sangeetha, R.; Arulpandi, I.; Geetha, A.

    2014-01-01

    Proteolysis-resistant lipases can be well exploited by industrial processes which employ both lipase and protease as biocatalysts. A proteolysis resistant lipase from Bacillus pumilus SG2 was isolated, purified and characterized earlier. The lipase was resistant to native and commercial proteases. In the present work, we have characterized the lip gene which encodes the proteolysis-resistant lipase from Bacillus pumilus SG2. The parameters and structural details of lipase were analysed. The lip gene consisted of 650 bp. The experimental molecular weight of SG2 lipase was nearly double that of its theoretical molecular weight, thus suggesting the existence of the functional lipase as a covalent dimer. The proteolytic cleavage sites of the lipase would have been made inaccessible by dimerisation, thus rendering the lipase resistant to protease. PMID:25242920

  5. Molecular characterization of a proteolysis-resistant lipase from Bacillus pumilus SG2.

    PubMed

    Sangeetha, R; Arulpandi, I; Geetha, A

    2014-01-01

    Proteolysis-resistant lipases can be well exploited by industrial processes which employ both lipase and protease as biocatalysts. A proteolysis resistant lipase from Bacillus pumilus SG2 was isolated, purified and characterized earlier. The lipase was resistant to native and commercial proteases. In the present work, we have characterized the lip gene which encodes the proteolysis-resistant lipase from Bacillus pumilus SG2. The parameters and structural details of lipase were analysed. The lip gene consisted of 650 bp. The experimental molecular weight of SG2 lipase was nearly double that of its theoretical molecular weight, thus suggesting the existence of the functional lipase as a covalent dimer. The proteolytic cleavage sites of the lipase would have been made inaccessible by dimerisation, thus rendering the lipase resistant to protease. PMID:25242920

  6. [Molecular docking of Bacillus pumilus xylanase and xylan substrate using computer modeling].

    PubMed

    Lin, Jin-Xia; Zhang, Liao-Yuan; Zhang, Guang-Ya; Fang, Bai-Shan

    2007-07-01

    Bacillus pumilus xylanase was cloned and sequenced. Based on the tertiary structure that originated from homology modeling, the potential active pocket was searched and ligand-protein docking was performed using relative softwares. The information extracted from the molecular docking is analyzed; several amino acid residues might play a vital role in the xylanase catalytic reaction are obtained to instruct the further modification of xylanase directed-evolution. PMID:17822050

  7. Complete genome sequence of Bacillus pumilus W3: A strain exhibiting high laccase activity.

    PubMed

    Guan, Zheng-Bing; Cai, Yu-Jie; Zhang, Yan-Zhou; Zhao, Hong; Liao, Xiang-Ru

    2015-08-10

    Here we report the full genome sequence of Bacillus pumilus W3, which was isolated from raw gallnut honey in Nandan County, Guangxi Province of China, showing high CotA-laccase activity. The W3 strain contains 3,745,123bp with GC content of 41.39%, and contains 3695 protein-coding genes, 21 rRNAs and 70 tRNAs. PMID:25957807

  8. Probiotic Bacillus pumilus SE5 shapes the intestinal microbiota and mucosal immunity in grouper Epinephelus coioides.

    PubMed

    Yang, Hong-Ling; Xia, Han-Qin; Ye, Yi-Dan; Zou, Wen-Chao; Sun, Yun-Zhang

    2014-09-30

    The health benefits of probiotics are thought to occur, at least in part, through an improved intestinal microbial balance in fish, although the molecular mechanisms whereby probiotics modulate the intestinal microbiota by means of activation of mucosal immunity are rarely explored. In this study, the effects of viable and heat-inactivated probiotic Bacillus pumilus SE5 on the intestinal dominant microbial community and mucosal immune gene expression were evaluated. The fish were fed for 60 d with 3 different diets: control (without probiotic), and diets T1 and T2 supplemented with 1.0 × 10? cells g?¹ viable and heat-inactivated B. pumilus SE5, respectively. Upregulated expression of TLR1, TLR2 and IL-8, but not MyD88 was observed in fish fed the viable probiotic, while elevated expression of TLR2, IL-8 and TGF-?1, but not MyD88 was observed in fish fed the heat-inactivated B. pumilus SE5. The induced activation of intestinal mucosal immunity, especially the enhanced expression of antibacterial epinecidin-1, was consistent with the microbial data showing that several potentially pathogenic bacterial species such as Psychroserpens burtonensis and Pantoea agglomerans were suppressed by both the viable and heat-inactivated probiotic B. pumilus SE5. These results lay the foundation for future studies on the molecular interactions between probiotics, intestinal microbiota and mucosal immunity in fish. PMID:25266899

  9. Antibacterial metabolites and bacteriolytic enzymes produced by Bacillus pumilus during bacteriolysis of Arthrobacter citreus.

    PubMed

    Brack, Christiane; Mikolasch, Annett; Schlueter, Rabea; Otto, Andreas; Becher, Dörte; Wegner, Uwe; Albrecht, Dirk; Riedel, Katharina; Schauer, Frieder

    2015-06-01

    The marine isolate Bacillus pumilus SBUG 1800 is able to lyse living cells of Arthrobacter citreus on solid media as well as pasteurized A. citreus cells in liquid mineral salt medium. The cultivation of B. pumilus in the presence of pasteurized A. citreus is accompanied by an enhanced production of 2,5-diketopiperazines (DKPs). DKPs inhibit bacterial growth, but do not seem to cause bacteriolysis. This study shows that B. pumilus also lyses living cells of A. citreus in co-culture experiments as an intraguild predator, even if the inoculum of B. pumilus is low. In order to characterize the bacteriolytic process, more precisely changes in the extracellular metabolome and proteome have been analyzed under different culture conditions. Besides the known DKPs, a number of different pumilacidins and bacteriolytic enzymes are produced. Two lipopeptides with [M?+?H](+)?=?1008 and [M?+?H](+)?=?1022 were detected and are proposed to be pumilacidin H and I. While the lipopeptides lyse living bacterial cells in lysis test assays, a set of extracellular enzymes degrades the dead cell material. Two of the cell wall hydrolases involved have been identified as N-acetylmuramoyl-L-alanine amidase and beta-N-acetylglucosaminidase. These findings together with electron microscopic and cell growth monitoring during co-culture experiments give a detailed view on the bacteriolytic process. PMID:25678259

  10. Selection of a Bacillus pumilus Strain Highly Active against Ceratitis capitata (Wiedemann) Larvae?

    PubMed Central

    Molina, C. Alfonso; Caña-Roca, Juan F.; Osuna, Antonio; Vilchez, Susana

    2010-01-01

    Ceratitis capitata (Wiedemann) (Diptera: Tephritidae), the Mediterranean fruit fly (medfly), is one of the most important fruit pests worldwide. The medfly is a polyphagous species that causes losses in many crops, which leads to huge economic losses. Entomopathogenic bacteria belonging to the genus Bacillus have been proven to be safe, environmentally friendly, and cost-effective tools to control pest populations. As no control method for C. capitata based on these bacteria has been developed, isolation of novel strains is needed. Here, we report the isolation of 115 bacterial strains and the results of toxicity screening with adults and larvae of C. capitata. As a result of this analysis, we obtained a novel Bacillus pumilus strain, strain 15.1, that is highly toxic to C. capitata larvae. The toxicity of this strain for C. capitata was related to the sporulation process and was observed only when cultures were incubated at low temperatures before they were used in a bioassay. The mortality rate for C. capitata larvae ranged from 68 to 94% depending on the conditions under which the culture was kept before the bioassay. Toxicity was proven to be a special characteristic of the newly isolated strain, since other B. pumilus strains did not have a toxic effect on C. capitata larvae. The results of the present study suggest that B. pumilus 15.1 could be considered a strong candidate for developing strategies for biological control of C. capitata. PMID:20038689

  11. Use of a plasmid DNA probe to monitor populations of Bacillus pumilus inoculant strains in hay

    SciTech Connect

    Hendrick, C.A.; Smiley, B.K.; Shelley, T.H.; Tomes, N.J. )

    1991-03-01

    The authors are evaluating naturally occurring isolates of Bacillus pumilus for use as microbial hay preservatives. Seven isolates of B, pumilus from hay contained a 42-kb cryptic plasmid (pMGD296). They wished to determine whether pMGD296 could be used as a molecular marker to follow populations of these isolates in hay over time. Southern blots and colony blots of 69 isolates of B. pumilus and other Bacillus spp. were probed with {sup 32}P-labeled pMGD296. Twenty-nine probe-positive isolates were identified; of these, 28 contained a plasmid with a restriction profile identical to that of pMGD296. One isolate from untreated hay contained a 40-kb plasmid (pMGD150) that was homologous to pMGD296 but had a different restriction fragment pattern. Regions of homology between the two plasmids were identified by Southern blotting, and a 1.9-kb HindIII-PstI fragment of pMGD296 lacking strong homology to pMGD150 was cloned in pUC18. The cloned fragment hybridized only with isolates containing pMGD296 and was used to estimate populations of these isolates in treated and untreated hay.

  12. Molecular characterization of cellulose-degrading Bacillus pumilus from the soil of tea garden, Darjeeling hills, India.

    PubMed

    Padaria, Jasdeep Chatrath; Sarkar, Kanishendranath; Lone, Showkat Ahmad; Srivastava, Sunita

    2014-05-01

    Bio-fuel produced from ethanol is economically and environmentally advantageous in context of changing global climate. A large number of microorganisms are capable of cellulase production but most of them cannot be utilized commercially due to their low activity. In the present study, an effiecient cellulose degrading strain of Bacillus pumilus was obtained after thorough screening for the production of extracellular cellulases. Out of a total of 144 microbes isolated from soils of Darjeeling hills of India, nineteen were found to be cellulose degrader under in vitro conditions as observed by clearing zone on CMC - agar plates. Isolate #35 had high cellulolytic activity as observed by a clearing zone of 26.83 mm diameter formed on CMC - agar plate. The isolate was characterized and identified as Bacillus pumilus. The isolate was submitted to National Agriculturally Important Microbial Culture Collection (NAIMCC), NBAIM, Mau with Accession number NAIMCC-B-01415. Transposon (Tn5) mutants of wild type isolate Bacillus pumilus NAIMCC-B-01415 were generated and screened for the absence of cellulose degradation. Of 365 B. pumilus NAIMCC-B-01415 mutants obtained, only two were unable to degrade cellulose under in vitro conditions. Inverse PCR studies with B. pumilus NAIMCC-B-01415 :: TL5, a cellulose degradation mutant of B. pumilus NAIMCC -B-01415 revealed presence of Cys B (Cystein protein regulatory) gene involved in cellulose degradation. The participation of Cys B gene in cellulase degradation is reported here. PMID:24813013

  13. Molecular cloning of enantioselective ester hydrolase from Bacillus pumilus DBRL-191.

    PubMed

    Rasool, Shafaq; Johri, Sarojini; Riyaz-ul-Hassan, Syed; Maqbool, Qurrat-ul-Ain; Verma, Vijeshwar; Koul, Surrinder; Taneja, Subhash C; Qazi, Ghulam N

    2005-08-01

    A gene from Bacillus pumilus expressed under its native promoter was cloned in Escherichia coli. Recombinant B. pumilus esterase (BPE) affects the kinetic resolution of racemic mixtures such as unsubstituted and substituted 1-(phenyl)ethanols (E approximately 33-103), ethyl 3-hydroxy-3-phenylpropanoate (E approximately 45-71), trans-4-fluorophenyl-3-hydroxymethyl-N-methylpiperidine (E approximately 10-13) and ethyl 2-hydroxy-4-phenylbutyrate (E approximately 7). The enzyme is composed of a 34-amino acid signal peptide and a 181-amino acid mature protein corresponding to a molecular weight of approximately 19.2kD and pI approximately 9.4. 3-D the structural model of the enzyme built by homology modelling using the atomic coordinates from the crystal structure of B. subtilis lipase (LipA) showed a compact minimal alpha/beta hydrolase fold. PMID:16006072

  14. Generation of biologically contained, readily transformable, and genetically manageable mutants of the biotechnologically important Bacillus pumilus.

    PubMed

    Wemhoff, Stephanie; Meinhardt, Friedhelm

    2013-09-01

    Bacillus pumilus mutants were generated by targeted deletion of a set of genes eventually facilitating genetic handling and assuring biological containment. The well-defined and stable mutants do not form functional endospores due to the deletion of yqfD, an essential sporulation gene; they are affected in DNA repair, as ?uvrBA rendered them UV hypersensitive and, thus, biologically contained; they are deficient for the uracil phosphoribosyl-transferase (?upp), allowing for 5-fluorouracil-based counterselection facilitating rapid allelic exchanges; and they are readily transformable due to the deletion of the restrictase encoding locus (?hsdR) of a type I restriction modification system. Vegetative growth as well as extracellular enzyme production and secretion are in no case affected. The combination of such gene deletions allows for development of B. pumilus strains suited for industrial use and further improvements. PMID:23644770

  15. Isolation, purification and characterisation of low molecular weight xylanase from Bacillus pumilus SSP-34.

    PubMed

    Subramaniyan, S

    2012-04-01

    Low molecular weight endo-xylanase from Bacillus pumilus SSP-34 was purified to homogeneity using ion exchange and size exclusion chromatographies. Xylanases were isolated by novel purification protocol which includes the use of anion exchange matrix such as DEAE Sepharose CL 6B with less affinity towards enzyme protein. The purified B. pumilus SSP-34 have a molecular weight of 20 kDa, with optimum pH and temperature at 6.0 and 50 °C, respectively. The enzyme was stable at 50 °C for 30 min. It showed remarkable stability at pH values ranging from 4.5 to 9 when the reaction was carried out at 50 °C. K (m) and V (max) values, determined with oats spelts xylan were 6.5 mg ml?¹ and 1,233 ?mol min?¹ mg?¹ protein, respectively, and the specific activity was 1,723 U mg?¹. PMID:22350873

  16. A new organic solvent tolerant protease from Bacillus pumilus 115b.

    PubMed

    Rahman, Raja Noor Zaliha Raja Abd; Mahamad, Shalihah; Salleh, Abu Bakar; Basri, Mahiran

    2007-07-01

    Five out of the nine benzene-toulene-ethylbenzene-xylene (BTEX) tolerant bacteria that demonstrated high protease activity on skim milk agar were isolated. Among them, isolate 115b identified as Bacillus pumilus exhibited the highest protease production. The protease produced was stable in 25% (v/v) benzene and toluene and it was activated 1.7 and 2.5- fold by n-dodecane and n-tetradecane, respectively. The gene encoding the organic solvent tolerant protease was cloned and its nucleotide sequence determined. Sequence analysis revealed an open reading frame (ORF) of 1,149 bp that encoded a polypeptide of 383 amino acid residues. The polypeptide composed of 29 residues of signal peptide, a propeptide of 79 residues and a mature protein of 275 amino acids with a calculated molecular mass of 27,846 Da. This is the only report available to date on organic solvent tolerant protease from B. pumilus. PMID:17492323

  17. Genotyping and toxigenic potential of Bacillus subtilis and Bacillus pumilus strains occurring in industrial and artisanal cured sausages.

    PubMed

    Matarante, Alessandra; Baruzzi, Federico; Cocconcelli, Pier Sandro; Morea, Maria

    2004-09-01

    Artisanal and industrial sausages were analyzed for their aerobic, heat-resistant microflora to assess whether new emerging pathogens could be present among Bacillus strains naturally contaminating cured meat products. Sixty-four isolates were characterized by randomly amplified polymorphic DNA (RAPD)-PCR and fluorescent amplified fragment length polymorphism (fAFLP). The biotypes, identified by partial 16S rRNA gene sequence analysis, belonged to Bacillus subtilis, Bacillus pumilus, and Bacillus amyloliquefaciens species. Both RAPD-PCR and fAFLP analyses demonstrated that a high genetic heterogeneity is present in the B. subtilis group even in strains harvested from the same source, making it possible to isolate 56 different biotypes. Moreover, fAFLP analysis made it possible to distinguish B. subtilis from B. pumilus strains. The strains were characterized for their toxigenic potential by molecular, physiological, and immunological techniques. Specific PCR analyses revealed the absence of DNA sequences related to HBL, BcET, NHE, and entFM Bacillus cereus enterotoxins and the enzymes sphingomyelinase Sph and phospholipase PI-PLC in all strains; also, the immunological analyses showed that Bacillus strains did not react with NHE- and HBL-specific antibodies. However, some isolates were found to be positive for hemolytic and lecithinase activity. The absence of toxigenic potential in Bacillus strains from the sausages analyzed indicates that these products can be considered safe under the processing conditions they were produced; however, great care should be taken when the ripening time is shortened, particularly in the case of traditional sausages, which could contain high amounts of Bacillus strains and possibly some B. cereus cells. PMID:15345396

  18. Biocontrol of Rhizoctonia solani damping-off disease in cucumber with Bacillus pumilus SQR-N43.

    PubMed

    Huang, Xinqi; Zhang, Nan; Yong, Xiaoyu; Yang, Xingming; Shen, Qirong

    2012-03-20

    Biological control is an efficient and environmentally friendly way to prevent damping-off disease. Micrographs were used to investigate the ability of Bacillus pumilus (B. pumilus) SQR-N43 to control Rhizoctonia solani (R. solani) Q1 in cucumbers. The root colonization ability of B. pumilus SQR-N43 was analyzed in vivo with a green fluorescent protein (GFP) tag. A pot experiment was performed to assess the in vivo disease-control efficiency of B. pumilus SQR-N43 and its bio-organic fertilizer. Results indicate that B. pumilus SQR-N43 induced hyphal deformation, enlargement of cytoplasmic vacuoles and cytoplasmic leakage in R. solani Q1 mycelia. A biofilm on the root surface was formed when the roots were inoculated with 10(7)-10(8)cells g(-1) of soil of GFP-tagged B. pumilus SQR-N43. In the pot experiment, the biocontrol reduced the concentration of R. solani. In contrast to applications of only B. pumilus SQR-N43 (N treatment), which produced control efficiencies of 23%, control efficiencies of 68% were obtained with applications of a fermented organic fertilizer inoculated with B. pumilus SQR-N43 (BIO treatment). After twenty days of incubation, significant differences in the number of CFUs and the percentage of spores of B. pumilus SQR-N43 were recorded between the N treatment (2.20×10(7)CFU g(-1) of soil and 79%, respectively) and the BIO treatment (1.67×10(8)CFU g(-1) of soil and 52%, respectively). The results indicate that B. pumilus SQR-N43 is a potent antagonist against R. solani Q1. The BIO treatment was more effective than the N treatment because it stabilized the population and increased the active form of the antagonist. PMID:21775112

  19. Characterization of cis-acting elements residing in the chitinase promoter of Bacillus pumilus SG2.

    PubMed

    Heravi, K Morabbi; Shali, A; Naghibzadeh, N; Ahmadian, G

    2014-05-01

    Bacillus pumilus SG2 is a chitinolytic bacterium that produces two chitinases, namely ChiS and ChiL. The chiS and chiL genes are consecutively expressed under a common promoter. Regulation of the chiS and chiL genes is under the control of carbon catabolite repression (CCR) in B. pumilus. This study aimed to investigate the cis-acting elements of the chitinase promoter. For this purpose, we transferred the chiS gene along with its specific promoter to Bacillus subtilis as a host. Primer extension analysis revealed two transcription start sites located 287 and 65 bp upstream of the chiS start codon. The distal promoter was highly compatible with the consensus sequence of the ?(A)-type promoters in B. subtilis, whereas the proximal promoter sequence showed less similarity to the ?(A)-type consensus sequence. A catabolite responsive element (cre), which is required for CCR in Bacillus species, was found to be 136 to 123 bp upstream of the chiS start codon. Interestingly, this cre site was located upstream of the -35 of the proximal promoter and downstream of the distal promoter. Deletion of this cre site sequence rendered the chiS expression constitutive. PMID:24293243

  20. Bacillus pumilus polysaccharide cross-reactive with meningococcal group A polysaccharide.

    PubMed

    Vann, W F; Liu, T Y; Robbins, J B

    1976-06-01

    A polysaccharide, antigenically and structurally related to meningococcal group A polysaccharide, was isolated from Bacillus pumilus Sh-17. This enteric bacterium has been implicated as a source of natural meningococcal group A immunity (Myerowitz et al., 1973). The B. pumilus polysaccharide was composed of a homopolymer of (1-6)-N-acetyl-manosamine-1-phosphate, glycerol phosphate teichoic acid-containing N-acetylglucosamine and alkali-labile alanine esters, and a mucopeptide. The cross-reaction was due to the poly-(1-6)-N-acetyl-mannosamine-1-phosphate in the B. pumilus and the meningococcal group A polysaccharides, based on the following evidence. Both polysaccharides contained N-acetyl-mannosamine phosphate. Periodate oxidized the mannosamine phosphate residues of the polysaccharide and destroyed their precipitating activity with meningococcal group A antiserum. Mild acid treatment released phosphomonoesters and destroyed the meningococcal group A precipitating activity of both polysaccharides. N-acetyl-mannosamine-6-phosphate inhibited the precipitation reaction between strain Sh-17 and meningococcal group A antisera. Only mannosamine phosphate was detected in trichloroacetic acid extracts of Sh-17 polysaccharide and meningococcal group A antigen-antibody precipitates. PMID:184043

  1. Bacillus pumilus laccase: a heat stable enzyme with a wide substrate spectrum

    PubMed Central

    2011-01-01

    Background Laccases are multi-copper oxidases that catalyze the one electron oxidation of a broad range of compounds. Laccase substrates include substituted phenols, arylamines and aromatic thiols. Such compounds are activated by the enzyme to the corresponding radicals. Owing to their broad substrate range laccases are considered to be versatile biocatalysts which are capable of oxidizing natural and non-natural industrial compounds, with water as sole by-product. Results A novel CotA-type laccase from Bacillus pumilus was cloned, expressed and purified and its biochemical characteristics are presented here. The molecular weight of the purified laccase was estimated to be 58 kDa and the enzyme was found to be associated with four copper atoms. Its catalytic activity towards 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid) (ABTS), 2,6-dimethoxyphenol (2,6-DMP) and syringaldazine (SGZ) was investigated. The kinetic parameters KM and kcat for ABTS were 80 ± 4 ?M and 291 ± 2.7 s-1, for 2,6-DMP 680 ± 27 ?M and 11 ± 0.1 s-1 and for SGZ only kcat could be estimated to be 66 ± 1.5 s-1. The pH optimum for ABTS was 4, for 2,6-DMP 7 and for SGZ 6.5 and temperature optima for ABTS and 2,6-DMP were found to be around 70°C. The screening of 37 natural and non-natural compounds as substrates for B. pumilus laccase revealed 18 suitable compounds. Three of them served as redox mediators in the laccase-catalyzed decolorization of the dye indigocarmine (IC), thus assessing the new enzyme's biotechnological potential. Conclusions The fully copper loaded, thermostable CotA laccase from Bacillus pumilus is a versatile laccase with potential applications as an industrial biocatalyst. PMID:21266052

  2. Chitinolytic and antifungal activity of a Bacillus pumilus chitinase expressed in Arabidopsis.

    PubMed

    Dehestani, Ali; Kazemitabar, Kamal; Ahmadian, Gholamreza; Jelodar, Nadali Babaeian; Salmanian, Ali Hatef; Seyedi, Mehdi; Rahimian, Heshmat; Ghasemi, Seyedhadi

    2010-04-01

    The Bacillus pumilus SG2 chitinase gene (ChiS) and its truncated form lacking chitin binding (ChBD) and fibronectin type III (FnIII) domains were transformed to Arabidopsis plants and the expression, functionality and antifungal activity of the recombinant proteins were investigated. Results showed that while the two enzyme forms showed almost equal hydrolytic activity toward colloidal chitin, they exhibited a significant difference in antifungal activity. Recombinant ChiS in plant protein extracts displayed a high inhibitory effect on spore germination and radial growth of hyphae in Alternaria brassicicola, Fusarium graminearum and Botrytis cinerea, while the activity of the truncated enzyme was strongly abolished. These findings demonstrate that ChBD and FnIII domains are not necessary for hydrolysis of colloidal chitin but play an important role in hydrolysis of chitin-glucan complex of fungal cell walls. Twenty microgram aliquots of protein extracts from ChiS transgenic lines displayed strong antifungal activity causing up to 80% decrease in fungal spore germination. This is the first report of a Bacillus pumilus chitinase expressed in plant system. PMID:20035370

  3. Characterization of a cryptic plasmid pPZZ84 from Bacillus pumilus.

    PubMed

    Zhang, Zhen-Hua; Tian, Wei; Liu, Dong-Yang; Liu, Yi-Chen; Shen, Qi-Rong; Shen, Biao

    2010-11-01

    The complete nucleotide sequence of a cryptic plasmid pPZZ84 from Bacillus pumilus strain ZZ84 was determined. Plasmid pPZZ84 is 6817bp long with GC content of 36.7%. Seven putative open reading frames were identified. ORF7 shows 91% and 90% amino acid identity with rep proteins of pSH1452 and pPL1, respectively, members of rolling-circle replication (RCR) pC194-family. A typical pC194-family double strand origin (dso), a single-stranded origin (sso) and rap (regulator aspartate phosphatase) proteins were also identified in the plasmid. These results imply that pPZZ84 belongs to the Bacillus subtilis species group of small rolling circle (BsSRC) replicating plasmids. The plasmid copy number of pPZZ84 in B. pumilus ZZ84 was estimated to be 46 per cell, more than that of other BsSRC plasmids in their hosts. PMID:20620162

  4. Genomic analysis of a ginger pathogen Bacillus pumilus providing the understanding to the pathogenesis and the novel control strategy

    PubMed Central

    Yuan, Yihui; Gao, Meiying

    2015-01-01

    Bacillus pumilus has been widely identified as a pathogen of plant and human, while the genetic information is rarely available for pathogenic B. pumilus strains. B. pumilus GR8 is a pathogen that causes ginger rhizome rot disease by invading ginger rhizome parenchymatous tissues, growing in the extracellular space, and producing plant cell wall-degrading enzymes to destroy ginger cells. In this study, the genome of GR8 was sequenced and characterized. This genome was the third completely sequenced genome of the B. pumilus species, and it exhibited high similarity to the genome of the B. pumilus strain B6033. The genome of GR8 was 3.67 Mb in length and encoded 3,713 putative ORFs. Among these predicted proteins, numerous plant cell wall-degrading enzymes and several proteins associated with invading and adapting to the environment in the extracellular space of the ginger rhizome parenchymatous tissue were found. The GR8 genome contained only one restriction-modification system and no CRISPR/Cas system. The lack of phage-resistant system suggested that phages might be potential agents for the control of GR8. The genomic analysis of GR8 provided the understanding to the pathogenesis and the phage-control strategy of pathogenic B. pumilus strains. PMID:25989507

  5. Genomic analysis of a ginger pathogen Bacillus pumilus providing the understanding to the pathogenesis and the novel control strategy.

    PubMed

    Yuan, Yihui; Gao, Meiying

    2015-01-01

    Bacillus pumilus has been widely identified as a pathogen of plant and human, while the genetic information is rarely available for pathogenic B. pumilus strains. B. pumilus GR8 is a pathogen that causes ginger rhizome rot disease by invading ginger rhizome parenchymatous tissues, growing in the extracellular space, and producing plant cell wall-degrading enzymes to destroy ginger cells. In this study, the genome of GR8 was sequenced and characterized. This genome was the third completely sequenced genome of the B. pumilus species, and it exhibited high similarity to the genome of the B. pumilus strain B6033. The genome of GR8 was 3.67 Mb in length and encoded 3,713 putative ORFs. Among these predicted proteins, numerous plant cell wall-degrading enzymes and several proteins associated with invading and adapting to the environment in the extracellular space of the ginger rhizome parenchymatous tissue were found. The GR8 genome contained only one restriction-modification system and no CRISPR/Cas system. The lack of phage-resistant system suggested that phages might be potential agents for the control of GR8. The genomic analysis of GR8 provided the understanding to the pathogenesis and the phage-control strategy of pathogenic B. pumilus strains. PMID:25989507

  6. Draft Genome Sequence of the Entomopathogenic Bacterium Bacillus pumilus 15.1, a Strain Highly Toxic to the Mediterranean Fruit Fly Ceratitis capitata.

    PubMed

    García-Ramón, Diana C; Palma, Leopoldo; Berry, Colin; Osuna, Antonio; Vílchez, Susana

    2015-01-01

    We present the draft whole-genome sequence of the entomopathogenic Bacillus pumilus 15.1 strain that consists of 3,795,691 bp and 3,776 predicted protein-coding genes. This genome sequence provides the basis for understanding the potential mechanism behind the toxicity and virulence of B. pumilus 15.1 against the Mediterranean fruit fly. PMID:26404596

  7. Draft Genome Sequence of the Entomopathogenic Bacterium Bacillus pumilus 15.1, a Strain Highly Toxic to the Mediterranean Fruit Fly Ceratitis capitata

    PubMed Central

    García-Ramón, Diana C.; Palma, Leopoldo; Berry, Colin; Osuna, Antonio

    2015-01-01

    We present the draft whole-genome sequence of the entomopathogenic Bacillus pumilus 15.1 strain that consists of 3,795,691 bp and 3,776 predicted protein-coding genes. This genome sequence provides the basis for understanding the potential mechanism behind the toxicity and virulence of B. pumilus 15.1 against the Mediterranean fruit fly. PMID:26404596

  8. Bacillus pumilus S124A carboxymethyl cellulase; a thermo stable enzyme with a wide substrate spectrum utility.

    PubMed

    Balasubramanian, Natesan; Simões, Nelson

    2014-06-01

    Bacillus pumilus S124A was identified as carboxymethyl cellulase producing bacteria from Azorean Bacillus collection (Lab collection), which was isolated in local soils. The bacterium was identified by 16S rRNA sequence and designated as Bacillus pumilus S124A. NCBI-blast analysis showed B. pumilus S124A; 16S rRNA sequence has high identity to other B. pumilus strains. Phylogenetic analysis showed B. pumilus S124A close to B. pumilus LZBP14 strain. CMcellulase was purified from cells-free supernatants and post mano-Q purification; 5.39% protein folds, and 0.88% recoveries were obtained. SDS-PAGE analysis showed molecular weight of the purified CMcellulase was estimated ?40kDa and composed of a single subunit. NonoLC ESI-MS/MS analysis was yielded four peptides, and protein has identity to other cellulases. Purified CMcellulase showed high activity to cellobiose followed by CMcellulose. Kinetic analysis showed Km, and Vmax were determined as 2.12mg/ml, 239?mol/min/mg, respectively. Optimum temperature and pH for the purified CMcellulase activity were found at 50°C and pH 6.0, respectively. Purified CMcellulase was maintained about 75% activity in a pH range of 4-8 and 70% activity in a temperature range of 40-70°C. CMcellulase activity was highly reduced by HgCl2, followed by EDTA, PMSF whereas CoCl2 was activated CMcellulase activity. PMID:24657377

  9. CynD, the Cyanide Dihydratase from Bacillus pumilus: Gene Cloning and Structural Studies

    PubMed Central

    Jandhyala, Dakshina; Berman, Mark; Meyers, Paul R.; Sewell, B. Trevor; Willson, Richard C.; Benedik, Michael J.

    2003-01-01

    The cyanide dihydratase in Bacillus pumilus was shown to be an 18-subunit spiral structure by three-dimensional reconstruction of electron micrographs of negatively stained material at its optimum pH, 8.0. At pH 5.4, the subunits rearrange to form an extended left-handed helix. Gel electrophoresis of glutaraldehyde cross-linked enzyme suggests that the fundamental component of the spiral is a dimer of the 37-kDa subunit. The gene was cloned, and the recombinant enzyme was readily expressed at high levels in Escherichia coli. Purification of the recombinant enzyme was facilitated by the addition of a C-terminal six-histidine affinity purification tag. The tagged recombinant enzyme has Km and Vmax values similar to those published for the native enzyme. This is the first cyanide dihydratase from a gram-positive bacterium to be sequenced, and it is the first description of the structure of any member of this enzyme class. The putative amino acid sequence shares over 80% identity to the only other sequenced cyanide dihydratase, that of the gram-negative Pseudomonas stutzeri strain AK61, and is similar to a number of other bacterial and fungal nitrilases. This sequence similarity suggests that the novel short spiral structure may be typical of these enzymes. In addition, an active cyanide dihydratase from a non-cyanide-degrading isolate of B. pumilus (strain 8A3) was cloned and expressed. This suggests that cynD, the gene coding for the cyanide dihydratase, is not unique to the C1 strain of B. pumilus and is not a reflection of its origin at a mining waste site. PMID:12902273

  10. CynD, the cyanide dihydratase from Bacillus pumilus: gene cloning and structural studies.

    PubMed

    Jandhyala, Dakshina; Berman, Mark; Meyers, Paul R; Sewell, B Trevor; Willson, Richard C; Benedik, Michael J

    2003-08-01

    The cyanide dihydratase in Bacillus pumilus was shown to be an 18-subunit spiral structure by three-dimensional reconstruction of electron micrographs of negatively stained material at its optimum pH, 8.0. At pH 5.4, the subunits rearrange to form an extended left-handed helix. Gel electrophoresis of glutaraldehyde cross-linked enzyme suggests that the fundamental component of the spiral is a dimer of the 37-kDa subunit. The gene was cloned, and the recombinant enzyme was readily expressed at high levels in Escherichia coli. Purification of the recombinant enzyme was facilitated by the addition of a C-terminal six-histidine affinity purification tag. The tagged recombinant enzyme has K(m) and V(max) values similar to those published for the native enzyme. This is the first cyanide dihydratase from a gram-positive bacterium to be sequenced, and it is the first description of the structure of any member of this enzyme class. The putative amino acid sequence shares over 80% identity to the only other sequenced cyanide dihydratase, that of the gram-negative Pseudomonas stutzeri strain AK61, and is similar to a number of other bacterial and fungal nitrilases. This sequence similarity suggests that the novel short spiral structure may be typical of these enzymes. In addition, an active cyanide dihydratase from a non-cyanide-degrading isolate of B. pumilus (strain 8A3) was cloned and expressed. This suggests that cynD, the gene coding for the cyanide dihydratase, is not unique to the C1 strain of B. pumilus and is not a reflection of its origin at a mining waste site. PMID:12902273

  11. Phylogenetic Diversity of the Bacillus pumilus Group and the Marine Ecotype Revealed by Multilocus Sequence Analysis

    PubMed Central

    Dong, Chunming; Sun, Fengqin; Wang, Liping; Li, Guangyu; Shao, Zongze

    2013-01-01

    Bacteria closely related to Bacillus pumilus cannot be distinguished from such other species as B. safensis, B. stratosphericus, B. altitudinis and B. aerophilus simply by 16S rRNA gene sequence. In this report, 76 marine strains were subjected to phylogenetic analysis based on 7 housekeeping genes to understand the phylogeny and biogeography in comparison with other origins. A phylogenetic tree based on the 7 housekeeping genes concatenated in the order of gyrB-rpoB-pycA-pyrE-mutL-aroE-trpB was constructed and compared with trees based on the single genes. All these trees exhibited a similar topology structure with small variations. Our 79 strains were divided into 6 groups from A to F; Group A was the largest and contained 49 strains close to B. altitudinis. Additional two large groups were presented by B. safensis and B. pumilus respectively. Among the housekeeping genes, gyrB and pyrE showed comparatively better resolution power and may serve as molecular markers to distinguish these closely related strains. Furthermore, a recombinant phylogenetic tree based on the gyrB gene and containing 73 terrestrial and our isolates was constructed to detect the relationship between marine and other sources. The tree clearly showed that the bacteria of marine origin were clustered together in all the large groups. In contrast, the cluster belonging to B. safensis was mainly composed of bacteria of terrestrial origin. Interestingly, nearly all the marine isolates were at the top of the tree, indicating the possibility of the recent divergence of this bacterial group in marine environments. We conclude that B. altitudinis bacteria are the most widely spread of the B. pumilus group in marine environments. In summary, this report provides the first evidence regarding the systematic evolution of this bacterial group, and knowledge of their phylogenetic diversity will help in the understanding of their ecological role and distribution in marine environments. PMID:24244618

  12. Use of Bacillus pumilus CBMAI 0008 and Paenibacillus sp. CBMAI 868 for colour removal from paper mill effluent

    PubMed Central

    de Oliveira, Patrícia Lopes; Duarte, Marta Cristina Teixeira; Ponezi, Alexandre Nunes; Durrant, Lúcia Regina

    2009-01-01

    Bacillus pumilus and Paenibacillus sp. were applied on the paper mill effluent to investigate the colour remotion. Inocula were individually applied in effluent at pH 7.0, 9.0 and 11.0. The real colour and COD remotion after 48h at pH 9.0 were, respectively, 41.87% and 22.08% for B. pumilus treatment and 42.30% and 22.89% for Paenibacillus sp. Gel permeation chromatography was used to verify the molar masses of compounds in the non-treated and treated effluent, showing a decrease in the compounds responsible for the paper mill effluent colour. PMID:24031372

  13. Optimization of chitin extraction from shrimp waste with Bacillus pumilus A1 using response surface methodology.

    PubMed

    Ghorbel-Bellaaj, Olfa; Hajji, Sawssen; Younes, Islem; Chaabouni, Moncef; Nasri, Moncef; Jellouli, Kemel

    2013-10-01

    Chitin extraction from shrimp shells by biological treatment, using the Bacilli Bacillus pumilus A1, is a non-polluting method and offers the opportunity to preserve the exceptional qualities of chitin and its derivatives. However, the major disadvantage of the fermentative way is the low efficiency of demineralization and deproteinization. The aim of this study is to improve the yield of extraction which depends on many factors, such as the medium composition and the physical parameters. In order to look for the optimal conditions, a Plackett and Burman design was carried out to screen eight factors influencing the deproteinization and demineralization efficiencies. The four most influencing variables were then examined to achieve the optimization using a central composite design. The results obtained showed that the optimal conditions were: shrimp shell concentration of 70 g/l, glucose concentration of 50 g/l, pH of 5.0 incubated with 0.225 OD of B. pumilus A1 inoculum, at 35 °C and 150 rpm for 6 days in 500 ml flask containing 100 ml of working volume. These conditions led to 88% of demineralization and 94% of deproteinization. (13)C CP/MAS NMR spectral analysis of the chitin prepared was carried out and was found to be similar to that of the commercial ?-chitin. PMID:23831901

  14. Metabolic flexibility of D-ribose producer strain of Bacillus pumilus under environmental perturbations.

    PubMed

    Srivastava, Rajesh K; Maiti, Soumen K; Das, Debasish; Bapat, Prashant M; Batta, Kritika; Bhushan, Mani; Wangikar, Pramod P

    2012-08-01

    The metabolic reaction rate vector is a bridge that links gene and protein expression alterations to the phenotypic endpoint. We present a simple approach for the estimation of flux distribution at key branch points in the metabolic network by using substrate uptake, metabolite secretion rate, and biomass growth rate for transketolase (tkt) deficient Bacillus pumilus ATCC 21951. We find that the glucose-6-phosphate (G6P) and pseudo catabolic/anabolic branch points are flexible in the D: -ribose-producing tkt deficient strain of B. pumilus. The normalized flux through the pentose phosphate pathway (PPP) varied from 1.5 to 86 % under different growth conditions, thereby enabling substantial extracellular accumulation of D: -ribose under certain conditions. Interestingly, the flux through PPP was affected by the extracellular phosphate concentration and dissolved oxygen concentration. This metabolic flexibility may have been the underlying reason for this strain being selected from thousands of others in a screening for D: -ribose producers conducted in the 1970s. PMID:22438109

  15. Purification and characterization of a novel collagenase from Bacillus pumilus Col-J.

    PubMed

    Wu, Qi; Li, Chen; Li, Chenglei; Chen, Hui; Shuliang, Liu

    2010-01-01

    The collagenase, produced extracellular by Bacillus pumilus Col-J, was purified by ammonium sulfate precipitation followed by two gel filtrations, involving Sephadex G-100 column and Sepharose Fast Flow column. Purified collagenase has a 31.53-fold increase in specific activity of 87.33 U/mg and 7.00% recovery. The collagenase has a relative molecular weight of 58.64 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The optimal temperature for the enzyme reaction was 45 degrees C. More than 50% of the original activity still remained after 5 min of incubation at 70 degrees C or 10 min at 60 degrees C. The maximal enzyme activity of collagenase was obtained at pH 7.5, and it was stable over a pH range of 6.5-8.0. The collagenase activity was strongly inhibited by Mn(2+), Pb(2+), ethylenediamine tetraacetic acid, ethylene glycol tetraacetic acid, and beta-mercaptoethanol. However, Ca(2+) and Mg(2+) greatly increased its activity. The collagenase from B. pumilus Col-J showed highly specific activity towards the native collagen from calf skin. The K(m) and V(max) of the enzyme for collagen were 0.79 mg/mL and 129.5 U, respectively. PMID:19475515

  16. Comparative effects of gamma rays and electron beams on spores of Bacillus pumilus

    SciTech Connect

    Hayashi, Toru; Todoriki, Setsuko ); Takizawa, Hironobu; Suzuki, Tetsuya; Takama, Kozo )

    1994-02-01

    The effects of [gamma] rays and electron beams on the germination, outgrowth and the synthesis of protein and RNA of Bacillus pumilus spores were investigated to clarify the difference in the effects of the two types of radiations on bacterial spores. Gamma irradiation facilitated the germination to a slightly larger degree than electron irradiation. The outgrowth, growth and the synthesis of protein and RNA were inhibited by [gamma] irradiation to a greater extent than electron irradiation, when the spores were irradiated at the same dose. However, the effects of the two types of radiations were the same when the spores were irradiated with electron beams at a dose 30% higher than [gamma] rays. The results indicate that the effects of electron beams on bacterial spores and those of [gamma] rays are qualitatively the same but quantitatively different. 23 refs., 5 figs.

  17. New derivatives of nonactic and homononactic acids from Bacillus pumilus derived from Breynia fruticosa.

    PubMed

    Han, Li; Huo, Peiyuan; Chen, Huahong; Li, Songtao; Jiang, Yi; Li, Liya; Xu, Lihua; Jiang, Chenglin; Huang, Xueshi

    2014-07-01

    Six new nonactic and homononactic acid derivatives, ethyl homononactate (1), ethyl nonactate (2), homononactyl homononactate (6), ethyl homononactyl nonactate (7), ethyl homononactyl homononactate (8), and ethyl nonactyl nonactate (9), as well as four known compounds, homononactic acid (3), nonactic acid (4), homononactyl nonactate (5), and bishomononactic acid (10), were isolated from culture broth of Bacillus pumilus derived from Breynia fruticosa. The structures of new compounds were elucidated by spectroscopic analysis and chemical methods. The optical purities of 1-6 were determined by HPLC/MS after treatment with L-phenylalanine methyl ester. The dimeric compounds 5-9 showed weak cytotoxic activities against five human cancer cell lines (IC50 19-100??g/ml). PMID:25044594

  18. Effects of oxygen free radical scavengers on the membrane myoinositol dehydrogenase of Bacillus pumilus strain 5.

    PubMed

    Eze, M O; Nnamani, L C; Ojiako, R I; Ogan, A U

    1987-01-01

    Micromolar amounts of superoxide dismutase (SOD) or parabenzoquinone (PBQ) inhibit the membrane-bound myoinositol dehydrogenase of Bacillus pumilus strain 5 in the mode of this enzyme transferring electrons to 2,6-dichlorophenol indophenol (DCPIP). The inhibition trends are similar to those reported earlier by us for the inhibition by mannitol and benzoate. We postulate that the transfer of electrons from the enzyme to DCPIP involves in its rate-limiting step, a catalytic intermediate in the nature of superoxide (O2-) and/or hydroyl free radical (OH.). Scavenging of any one or both of these radicals, therefore, inhibits the electron transfer reaction. PBQ serves as an electron sink in the reaction preventing the reduction of DCPIP. PMID:3508132

  19. Purification and characterization of an acetyl xylan esterase from Bacillus pumilus.

    PubMed

    Degrassi, G; Okeke, B C; Bruschi, C V; Venturi, V

    1998-02-01

    Bacillus pumilus PS213 was found to be able to release acetate from acetylated xylan. The enzyme catalyzing this reaction has been purified to homogeneity and characterized. The enzyme was secreted, and its production was induced by corncob powder and xylan. Its molecular mass, as determined by gel filtration, is 190 kDa, while sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed a single band of 40 kDa. The isoelectric point was found to be 4.8, and the enzyme activity was optimal at 55 degrees C and pH 8.0. The activity was inhibited by most of the metal ions, while no enhancement was observed. The Michaelis contant (Km) and Vmax for alpha-naphthyl acetate were 1.54 mM and 360 micromol min-1 mg of protein-1, respectively. PMID:10215579

  20. Sequence of the gene encoding an alkaline serine proteinase of Bacillus pumilus TYO-67.

    PubMed

    Aoyama, M; Toma, C; Yasud, M; Iwanaga, M

    2000-01-01

    The complete nucleotide sequence of the gene encoding an alkaline serine proteinase (aprP) of Bacillus pumilus TYO-67 was determined. The sequence analysis showed an open reading frame of 1,149 bp (383 amino acids) that encoded a signal peptide consisting of 29 residues and a propeptide of 79 residues. The deduced 3 amino acid residues, D32, H64, and S221, were identical with 3 essential amino acids in the catalytic center of subtilases. The sequence around these residues revealed that APRP was a new member of the true subtilisin subgroup of the subtilisin family. The highest homology was found in subtilisin NAT at 64.4% in the DNA sequence. The residue S189 of APRP was different from those of other subtilases. PMID:10888358

  1. Characterization and a point mutational approach of a psychrophilic lipase from an arctic bacterium, Bacillus pumilus.

    PubMed

    Wi, Ah Ram; Jeon, Sung-Jong; Kim, Sunghui; Park, Ha Ju; Kim, Dockyu; Han, Se Jong; Yim, Joung Han; Kim, Han-Woo

    2014-06-01

    A bacterium with lipolytic activity was isolated from the Chukchi Sea within the Arctic Ocean. The lipase BpL5 from the isolate, Bacillus pumilus ArcL5, belongs to subfamily 4 of lipase family I. The optimum pH and temperature of the recombinant enzyme BpL5, as expressed in Escherichia coli, were 9.0 and 20 °C, respectively. The enzyme retained 85 % of its activity at 5 °C. There was a significant difference between temperatures for maximal activity (20 °C) and for protein denaturation (approx. 45 °C). The enzyme preferred middle-chain (C8) p-nitrophenyl substrates. Two mutants, S139A and S139Y, were rationally designed based on the 3D-structure model, and their activities were compared with that of the wild type. The both mutants showed significantly improved activity against tricaprylin. PMID:24563306

  2. Purification and Characterization of an Acetyl Xylan Esterase from Bacillus pumilus

    PubMed Central

    Degrassi, Giuliano; Okeke, Benedict C.; Bruschi, Carlo V.; Venturi, Vittorio

    1998-01-01

    Bacillus pumilus PS213 was found to be able to release acetate from acetylated xylan. The enzyme catalyzing this reaction has been purified to homogeneity and characterized. The enzyme was secreted, and its production was induced by corncob powder and xylan. Its molecular mass, as determined by gel filtration, is 190 kDa, while sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed a single band of 40 kDa. The isoelectric point was found to be 4.8, and the enzyme activity was optimal at 55°C and pH 8.0. The activity was inhibited by most of the metal ions, while no enhancement was observed. The Michaelis constant (Km) and Vmax for ?-naphthyl acetate were 1.54 mM and 360 ?mol min?1 mg of protein?1, respectively. PMID:10215579

  3. Biological and genomic analysis of a PBSX-like defective phage induced from Bacillus pumilus AB94180.

    PubMed

    Jin, Tingting; Zhang, Xiaoming; Zhang, Yang; Hu, Zhongsheng; Fu, Zhengwei; Fan, Junpeng; Wu, Ming; Wang, Yi; Shen, Ping; Chen, Xiangdong

    2014-04-01

    Defective prophages, which are found in the genomes of many bacteria, are unable to complete a viral replication cycle and propagate in their hosts as healthy prophages. They package random DNA fragments derived from various sites of the host chromosome instead of their own genomes. In this study, we characterized a defective phage, PBP180, which was induced from Bacillus pumilus AB94180 by treatment with mitomycin C. Electron microscopy showed that the PBP180 particle has a head with a hexagonal outline of ~40 nm in diameter and a long tail. The DNA packaged in the PBP180 head consists of 8-kb DNA fragments from random portions of the host chromosome. The head and tail proteins of the PBP180 particle consist of four major proteins of approximately 49, 33, 16 and 14 kDa. The protein profile of PBP180 is different from that of PBSX, a well-known defective phage induced from Bacillus subtilis 168. A killing activity test against two susceptible strains each of B. subtilis and B. pumilus showed that the defective particles of PBP180 killed three strains other than its own host, B. pumilus AB94180, differing from the host-killing ranges of the defective phages PBSX, PBSZ (induced from B. subtilis W23), and PBSX4 (induced from B. pumilus AB94044). The genome of the PBP180 prophage, which is integrated in the B. pumilus AB94180 chromosome, is 28,205 bp in length, with 40 predicted open reading frames (ORFs). Further genomic comparison of prophages PBP180, PBSX, PBSZ and other PBSX-like prophage elements in B. pumilus strains revealed that their overall architectures are similar, but significant low homology exists in ORF29-ORF38, which presumably encode tail fiber proteins involved in recognition and killing of susceptible strains. PMID:24154951

  4. Expression and biochemical characterization of cold-adapted lipases from Antarctic Bacillus pumilus strains.

    PubMed

    Litantra, Ribka; Lobionda, Stefani; Yim, Joung Han; Kim, Hyung Kwoun

    2013-09-28

    Two lipase genes (bpl1 and bpl3) from Antarctic Bacillus pumilus strains were expressed in Bacillus subtilis. Both recombinant lipases BPL1 and BPL2 were secreted to the culture medium and their activities reached 3.5 U/ml and 5.0 U/ml, respectively. Their molecular masses apparent using SDS-PAGE were 23 kDa for BPL1 and 19 kDa for BPL3. Both lipases were purified to homogeneity using ammonium sulfate precipitation and HiTrap SP FF column and Superose 12 column chromatographies. The final specific activities were estimated to be 328 U/mg for BPL1 and 310 U/mg for BPL3. Both lipases displayed an optimum temperature of 35°C, similar to other mesophilic enzymes. However, they maintained as much as 70% and 80% of the maximum activities at 10°C. Accordingly, their calculated activation energy at a temperature range of 10-35°C was 5.32 kcal/mol for BPL1 and 4.26 kcal/mol for BPL3, typical of cold-adapted enzymes. The optimum pH of BPL1 and BPL3 was 8.5 and 8.0, respectively, and they were quite stable at pH 7.0-11.0, showing their strong alkaline tolerance. Both lipases had a preference toward medium chain length (C6-C10) fatty acid substrates. These results indicate the potential for the two Antarctic B. pumilus lipases as catalysts in bioorganic synthesis, food, and detergent industries. PMID:23770563

  5. Improving the chitinolytic activity of Bacillus pumilus SG2 by random mutagenesis.

    PubMed

    Vahed, Majid; Motalebi, Ebrahim; Rigi, Garshasb; Akbari Noghabi, Kambiz; Soudi, Mohammad Reza; Sadeghi, Mehdi; Ahmadian, Gholamreza

    2013-11-28

    Bacillus pumilus SG2, a halotolerant strain, expresses two major chitinases designated ChiS and ChiL that were induced by chitin and secreted into the supernatant. The present work aimed to obtain a mutant with higher chitinolytic activity through mutagenesis of Bacillus pumilus SG2 using a combination of UV irradiation and nitrous acid treatment. Following mutagenesis and screening on chitin agar and subsequent formation of halos, the mutated strains were examined for degradation of chitin under different conditions. A mutant designated AV2-9 was selected owing to its higher chitinase activity. To search for possible mutations in the whole operon including ChiS and ChiL, the entire chitinase operon, including the intergenic region, promoter, and two areas corresponding to the ChiS and ChiL ORF, was suquenced. Nucleotide sequence analysis of the complete chitinase operon from the SG2 and AV2-9 strains showed the presence of a mutation in the catalytic domain (GH18) of chitinase (ChiL). The results demonstrated that a single base change had occurred in the ChiL sequence in AV2- 9. The wild-type chitinase, ChiL, and the mutant (designated ChiLm) were cloned, expressed, and purified in E. coli. Both enzymes showed similar profiles of activity at different ranges of pH, NaCl concentration, and temperature, but the mutant enzyme showed approximately 30% higher catalytic activity under all the conditions tested. The results obtained in this study showed that the thermal stability of chitinase increased in the mutant strain. Bioinformatics analysis was performed to predict changes in the stability of proteins caused by mutation. PMID:23867702

  6. [Directed evolution of lipase of Bacillus pumilus YZ02 by error-prone PCR].

    PubMed

    Huang, Ying; Cai, Yong; Yang, Jiangke; Yan, Yunjun

    2008-03-01

    Random mutagenesis on Bacillus pumilus lipase YZ02 gene was conducted by using error-prone PCR strategy. Through two cycles of directed evolution, two optimum mutants BpL1-7 and BpL2-1369 with lipase activity improved 2 folds and 6 folds respectively were screened. The sequence of BpL2-1369 lipase gene showed that four nucleotides substitution, T61C, C147T, A334G and T371A have occurred, and three of them caused amino acid changes. Thus, amine acid Ser21 was changed into Pro21, Arg112 to Gly112, and Leu124 to His124. According to the 3D structure of Bacillus pumilus lipase mimicked by SWISS-MODEL Repository, three mutated amino acids were located at the third amino acid of the first alpha-helix, the turn between the fourth and fifth beta fold, and the first amino acid of the fifth beta fold, respectively. The BpL and BpL2-1369 genes were ligated into pET28a vector, and transferred into E. coli BL21 (DE3). After induced by IPTG the lipases were purified and characterized. The results showed that the specific activity of the evolved lipase was 1.31-fold than that of the wild lipase, and the Km decreased from 8.24 mmol/L to 7.17 mmol/L. The pH stability of the evolved lipase was better than wild lipase when pH>8.0. PMID:18589821

  7. Survival of Bacillus pumilus SAFR-032 in simulated Mars atmosphere in real space conditions

    NASA Astrophysics Data System (ADS)

    Vaishampayan, Parag A.; Fox, George E.; Venkateswaran, Kasthuri

    2010-09-01

    To prevent forward contamination and maintain the scientific integrity of future life detection missions, it is important to characterize and attempt to eliminate terrestrial microorganisms associated with exploratory spacecraft and landing vehicles. Among the organisms isolated from spacecraft-associated habitats, spores of Bacillus pumilus SAFR-032 exhibited unusually high resistance to decontamination techniques such as UVradiation and peroxide treatment. Subsequently, Bacillus pumilus SAFR-032 was flown to the International Space Station (ISS) and exposed to a variety of space conditions using the European Technology Exposure Platform and Experiment Facility (EuTEF). After 18 months exposure in the EuTEF facility under dark space conditions, SAFR-032 spores showed 10 to 40% survivability, whereas a survival rate of 85 to 100% was observed when these spores were kept aboard the ISS under dark simulated-Mars atmospheric conditions. In contrast, when UV (>110nm) was exerted on SAFR-032 spores for the same time period and conditions using the EuTEF, a ~7-log reduction in viability was noticed. However, the UV exposure still did not inactivate all the spores as 19 CFUs were later isolated via cultivation. A parallel experiment was conducted on Earth with identical samples but under simulated conditions. Spores exposed to ground simulations showed less of a reduction in viability when compared with the "real space" exposed spores (~3-log reduction in viability for Mars UV, and ~4-log reduction in viability for Space UV). The data generated is important to assess the probability and mechanisms of microbial survival, microbial contaminants of risk for forward contamination, in situ life detection, and to safeguard the integrity of sample return missions.

  8. Matrix Production, Pigment Synthesis, and Sporulation in a Marine Isolated Strain of Bacillus pumilus

    PubMed Central

    Di Luccia, Blanda; Riccio, Antonio; Vanacore, Adele; Baccigalupi, Loredana; Molinaro, Antonio; Ricca, Ezio

    2015-01-01

    The ability to produce an extracellular matrix and form multicellular communities is an adaptive behavior shared by many bacteria. In Bacillus subtilis, the model system for spore-forming bacteria, matrix production is one of the possible differentiation pathways that a cell can follow when vegetative growth is no longer feasible. While in B. subtilis the genetic system controlling matrix production has been studied in detail, it is still unclear whether other spore formers utilize similar mechanisms. We report that SF214, a pigmented strain of Bacillus pumilus isolated from the marine environment, can produce an extracellular matrix relying on orthologs of many of the genes known to be important for matrix synthesis in B. subtilis. We also report a characterization of the carbohydrates forming the extracellular matrix of strain SF214. The isolation and characterization of mutants altered in matrix synthesis, pigmentation, and spore formation suggest that in strain SF214 the three processes are strictly interconnected and regulated by a common molecular mechanism. PMID:26506360

  9. Purification and partial characterization of serine-metallokeratinase from a newly isolated Bacillus pumilus NRC21.

    PubMed

    Tork, Sanaa E; Shahein, Yasser E; El-Hakim, Amr E; Abdel-Aty, Azza M; Aly, Magda M

    2016-05-01

    A serine metallokeratinase enzyme (30kDa) produced by a newly isolated Bacillus strain (Bacillus pumilus NRC21) cultivated under optimized conditions in medium containing chicken feather meal was purified and characterized in a set of biochemical assays. The purification was carried out using two successive chromatographic steps; cation exchange chromatography on CM-cellulose and gel filtration on sephadex G-100 columns. The purified enzyme showed a specific activity of 2000units/mg protein against 170units/mg protein for crude extract with 12 fold purification. The enzymatic activity of the keratinase stimulated by (Na(+), K(+), Mg(2+)), Hg(+2) had no effect, and inhibited by entire tested cations, serine and metalloproteinase inhibitors, therefore it can be considered as a serine metalloenzyme. The optimum pH and temperature for the purified enzyme were (7.5, 8.5) and (50, 45°C) when using keratin azure and azocasein as substrates, respectively. The purified enzyme was highly stable at broad pH and temperature ranged (5-10) and (20-60°C), respectively and its thermoactivity and thermostability were enhanced in the presence of 5mMMg(+2). These results suggest that the purified keratinase may be used in several industrial applications. PMID:26802243

  10. Matrix Production, Pigment Synthesis, and Sporulation in a Marine Isolated Strain of Bacillus pumilus.

    PubMed

    Di Luccia, Blanda; Riccio, Antonio; Vanacore, Adele; Baccigalupi, Loredana; Molinaro, Antonio; Ricca, Ezio

    2015-10-01

    The ability to produce an extracellular matrix and form multicellular communities is an adaptive behavior shared by many bacteria. In Bacillus subtilis, the model system for spore-forming bacteria, matrix production is one of the possible differentiation pathways that a cell can follow when vegetative growth is no longer feasible. While in B. subtilis the genetic system controlling matrix production has been studied in detail, it is still unclear whether other spore formers utilize similar mechanisms. We report that SF214, a pigmented strain of Bacillus pumilus isolated from the marine environment, can produce an extracellular matrix relying on orthologs of many of the genes known to be important for matrix synthesis in B. subtilis. We also report a characterization of the carbohydrates forming the extracellular matrix of strain SF214. The isolation and characterization of mutants altered in matrix synthesis, pigmentation, and spore formation suggest that in strain SF214 the three processes are strictly interconnected and regulated by a common molecular mechanism. PMID:26506360

  11. An in-depth characterization of the entomopathogenic strain Bacillus pumilus 15.1 reveals that it produces inclusion bodies similar to the parasporal crystals of Bacillus thuringiensis.

    PubMed

    Garcia-Ramon, Diana C; Molina, C Alfonso; Osuna, Antonio; Vílchez, Susana

    2016-04-01

    In the present work, the local isolate Bacillus pumilus 15.1 has been morphologically and biochemically characterized in order to gain a better understanding of this novel entomopathogenic strain active against Ceratitis capitata. This strain could represent an interesting biothechnological tool for the control of this pest. Here, we report on its nutrient preferences, extracellular enzyme production, motility mechanism, biofilm production, antibiotic suceptibility, natural resistance to chemical and physical insults, and morphology of the vegetative cells and spores. The pathogen was found to be β-hemolytic and susceptible to penicillin, ampicillin, chloramphenicol, gentamicin, kanamycin, rifampicin, tetracycline, and streptomycin. We also report a series of biocide, thermal, and UV treatments that reduce the viability of B. pumilus 15.1 by several orders of magnitude. Heat and chemical treatments kill at least 99.9 % of vegetative cells, but spores were much more resistant. Bleach was the only chemical that was able to completely eliminate B. pumilus 15.1 spores. Compared to the B. subtilis 168 spores, B. pumilus 15.1 spores were between 2.67 and 350 times more resistant to UV radiation while the vegetative cells of B. pumilus 15.1 were almost up to 3 orders of magnitude more resistant than the model strain. We performed electron microscopy for morphological characterization, and we observed geometric structures resembling the parasporal crystal inclusions synthesized by Bacillus thuringiensis. Some of the results obtained here such as the parasporal inclusion bodies produced by B. pumilus 15.1 could potentially represent virulence factors of this novel and potentially interesting strain. PMID:26782747

  12. Survival of Bacillus pumilus spores for a prolonged period of time in real space conditions.

    PubMed

    Vaishampayan, Parag A; Rabbow, Elke; Horneck, Gerda; Venkateswaran, Kasthuri J

    2012-05-01

    To prevent forward contamination and maintain the scientific integrity of future life-detection missions, it is important to characterize and attempt to eliminate terrestrial microorganisms associated with exploratory spacecraft and landing vehicles. Among the organisms isolated from spacecraft-associated surfaces, spores of Bacillus pumilus SAFR-032 exhibited unusually high resistance to decontamination techniques such as UV radiation and peroxide treatment. Subsequently, B. pumilus SAFR-032 was flown to the International Space Station (ISS) and exposed to a variety of space conditions via the European Technology Exposure Facility (EuTEF). After 18 months of exposure in the EXPOSE facility of the European Space Agency (ESA) on EuTEF under dark space conditions, SAFR-032 spores showed 10-40% survivability, whereas a survival rate of 85-100% was observed when these spores were kept aboard the ISS under dark simulated martian atmospheric conditions. In contrast, when UV (>110?nm) was applied on SAFR-032 spores for the same time period and under the same conditions used in EXPOSE, a ?7-log reduction in viability was observed. A parallel experiment was conducted on Earth with identical samples under simulated space conditions. Spores exposed to ground simulations showed less of a reduction in viability when compared with the "real space" exposed spores (?3-log reduction in viability for "UV-Mars," and ?4-log reduction in viability for "UV-Space"). A comparative proteomics analysis indicated that proteins conferring resistant traits (superoxide dismutase) were present in higher concentration in space-exposed spores when compared to controls. Also, the first-generation cells and spores derived from space-exposed samples exhibited elevated UVC resistance when compared with their ground control counterparts. The data generated are important for calculating the probability and mechanisms of microbial survival in space conditions and assessing microbial contaminants as risks for forward contamination and in situ life detection. PMID:22680694

  13. Bacillus pumilus ES4: candidate plant growth-promoting bacterium to enhance establishment of plants in mine tailings

    PubMed Central

    de-Bashan, Luz E.; Hernandez, Juan-Pablo; Bashan, Yoav; Maier, Raina

    2014-01-01

    Three plant growth-promoting bacteria (PGPB; Bacillus pumilus ES4, B. pumilus RIZO1, and Azospirillum brasilense Cd) were tested for their ability to enhance plant growth and development of the native Sonoran Desert shrub quailbush (Atriplex lentiformis) and for their effect on the native bacterial community in moderately acidic, high-metal content (AHMT) and in neutral, low metal content natural tailings (NLMT) in controlled greenhouse experiments. Inoculation of quailbush with all three PGPB significantly enhanced plant growth parameters, such as germination, root length, dry weight of shoots and roots, and root/shoot ratio in both types of tailings. The effect of inoculation on the indigenous bacterial community by the most successful PGPB Bacillus pumilus ES4 was evaluated by denaturating gradient gel electrophoresis (PCR-DGGE) fingerprinting and root colonization was followed by specific fluorescent in situ hybridization (FISH). Inoculation with this strain significantly changed the bacterial community over a period of 60 days. FISH analysis showed that the preferred site of colonization was the root tips and root elongation area. This study shows that inoculation of native perennial plants with PGPB can be used for developing technologies for phytostabilizing mine tailings. PMID:25009362

  14. Isolation of a novel strain of Bacillus pumilus from penaeid shrimp that is inhibitory against marine pathogens.

    PubMed

    Hill, J E; Baiano, J C F; Barnes, A C

    2009-12-01

    A bacterium was isolated from the mid-gut of healthy black tiger shrimp, Penaeus monodon, based on a large zone of inhibition in mixed culture on solid medium. The isolate was a Gram-positive, motile spore former, with an optimum pH range for growth in tryptone soya broth containing 2% NaCl of between pH 6 and 9. The bacterium was highly salt tolerant with concentrations between 0% and 8% having no detrimental effect on growth. The isolate was identified as Bacillus pumilus based on physiological capabilities using the API50CHB and Biolog systems. Amplification and sequencing of the 16S rRNA gene followed by phylogenetic analysis confirmed its identity. The Bacillus pumilus isolate was strongly inhibitory against the marine bacterial pathogens Vibrio alginolyticus, V. mimicus and V. harveyi, and weakly inhibitory against V. parahaemolyticus in cross-streaking assays on solid medium. The organism was marginally self-inhibitory, and inhibited B. licheniformis and B. subtilis. The suitability of the B. pumilus isolate for use as a probiotic in farmed shrimp was further supported by the absence of any of the known B. cereus enterotoxin genes. Based on these in vitro results, in vivo safety and efficacy trials are underway to determine suitability of the novel strain as a commercial probiotic. PMID:19573134

  15. Catalytic, kinetic and thermodynamic properties of Bacillus pumilus FH9 keratinase conjugated with activated pectin.

    PubMed

    Abdel-Naby, Mohamed A; A Ibrahim, M H; El-Refai, H A

    2016-04-01

    Bacillus pumilus FH9 keratinase was covalently coupled to several oxidized polysaccharides. The conjugates were evaluated for the retained activity, kinetic and thermodynamic stability. Among all preparations, the conjugated enzyme with oxidized pectin had the highest recovered activity (71.75%) and the highest thermal stability at 60°C (t1/2=333min). Compared to the native enzyme, the conjugated preparation exhibited higher optimum temperature, lower activation energy (Ea), lower deactivation constant rate (kd), higher t1/2, and higher decimal reduction time values (D) within the temperature range of 50-80°C. The thermodynamic parameters (ΔH*, ΔG*, ΔS*) of irreversible thermal denaturation for the native and conjugated keratinase were also evaluated. The values of enthalpy of activation (ΔH*), free energy of activation (ΔG*), and free energy of transition state binding (ΔG*E-T) for keratin hydrolysis were lower for the conjugated enzyme. Moreover, there was highly significant impact on improving the values of Vmax/Km, kcat, kcat/Km, and ΔG*E-S for the modified enzyme. Both native and conjugated enzymes were slightly activated by CaCl2 and MgCl2. However, the inhibitory effects of EDTA, HgCl2 and ZnSO4 were more pronounced with the native enzyme. PMID:26743746

  16. Acetoin Catabolism and Acetylbutanediol Formation by Bacillus pumilus in a Chemically Defined Medium

    PubMed Central

    Xiao, Zijun; Ma, Cuiqing; Xu, Ping; Lu, Jian R.

    2009-01-01

    Background Most low molecular diols are highly water-soluble, hygroscopic, and reactive with many organic compounds. In the past decades, microbial research to produce diols, e.g. 1,3-propanediol and 2,3-butanediol, were considerably expanded due to their versatile usages especially in polymer synthesis and as possible alternatives to fossil based feedstocks from the bioconversion of renewable natural resources. This study aimed to provide a new way for bacterial production of an acetylated diol, i.e. acetylbutanediol (ABD, 3,4-dihydroxy-3-methylpentan-2-one), by acetoin metabolism. Methodology/Principal Findings When Bacillus pumilus ATCC 14884 was aerobically cultured in a chemically defined medium with acetoin as the sole carbon and energy source, ABD was produced and identified by gas chromatography – chemical ionization mass spectrometry and NMR spectroscopy. Conclusions/Significance Although the key enzyme leading to ABD from acetoin has not been identified yet at this stage, this study proposed a new metabolic pathawy to produce ABD in vivo from using renewable resources – in this case acetoin, which could be reproduced from glucose in this study – making it the first facility in the world to prepare this new bio-based diol product. PMID:19461961

  17. Antibiofilm activity of Bacillus pumilus SW9 against initial biofouling on microfiltration membranes.

    PubMed

    Zhang, Ying; Yu, Xin; Gong, Song; Ye, Chengsong; Fan, Zihong; Lin, Huirong

    2014-02-01

    Membrane biofouling, resulting from biofilm formation on the membrane, has become the main obstacle hindering wider application of membrane technology. Initial biofouling proves to be crucial which involves early stages of microbial adhesion and biofilm formation. Biological control of microbial attachment seems to be a promising strategy due to its high efficiency and eco-friendliness. The present study investigated the effects of a bacterium Bacillus pumilus SW9 on controlling the initial fouling formed by four target bacterial strains which were pioneer species responsible for biofouling in membrane bioreactors, using microfiltration membranes as the abiotic surfaces. The results suggested that strain SW9 exhibited excellent antibiofilm activity by decreasing the attached biomass of target strains. The production of extracellular polysaccharides and proteins by four target strains was also reduced. A distinct improvement of permeate flux in dead-end filtration systems was achieved when introducing strain SW9 to microfiltration experiments. Scanning electron microscopy and confocal laser scanning microscopy were performed to further ascertain significant changes of the biofouling layers. A link between biofilm inhibition and initial biofouling mitigation was thus provided, suggesting an alternatively potential way to control membrane biofouling through bacterial interactions. PMID:23715854

  18. Isolation and characterization of a cyanide dihydratase from Bacillus pumilus C1.

    PubMed Central

    Meyers, P R; Rawlings, D E; Woods, D R; Lindsey, G G

    1993-01-01

    A cyanide-degrading enzyme from Bacillus pumilus C1 has been purified and characterized. This enzyme consisted of three polypeptides of 45.6, 44.6, and 41.2 kDa; the molecular mass by gel filtration was 417 kDa. Electron microscopy revealed a multimeric, rod-shaped protein approximately 9 by 50 nm. Cyanide was rapidly degraded to formate and ammonia. Enzyme activity was optimal at 37 degrees C and pH 7.8 to 8.0. Activity was enhanced by Sc3+, Cr3+, Fe3+, and Tb3+; enhancement was independent of metal ion concentration at concentrations above 5 microM. Reversible enhancement of enzymatic activity by azide was maximal at 4.5 mM azide and increased with time. No activity was recorded with the cyanide substrate analogs CNO-, SCN-, CH3CN, and N3- and the possible degradation intermediate HCONH2. Kinetic studies indicated a Km of 2.56 +/- 0.48 mM for cyanide and a Vmax of 88.03 +/- 4.67 mmol of cyanide per min/mg/liter. The Km increased approximately twofold in the presence of 10 microM Cr3+ to 5.28 +/- 0.38 mM for cyanide, and the Vmax increased to 197.11 +/- 8.51 mmol of cyanide per min/mg/liter. We propose naming this enzyme cyanide dihydratase. Images PMID:8407782

  19. Probing an Interfacial Surface in the Cyanide Dihydratase from Bacillus pumilus, A Spiral Forming Nitrilase

    PubMed Central

    Park, Jason M.; Mulelu, Andani; Sewell, B. Trevor; Benedik, Michael J.

    2016-01-01

    Nitrilases are of significant interest both due to their potential for industrial production of valuable products as well as degradation of hazardous nitrile-containing wastes. All known functional members of the nitrilase superfamily have an underlying dimer structure. The true nitrilases expand upon this basic dimer and form large spiral or helical homo-oligomers. The formation of this larger structure is linked to both the activity and substrate specificity of these nitrilases. The sequences of the spiral nitrilases differ from the non-spiral forming homologs by the presence of two insertion regions. Homology modeling suggests that these regions are responsible for associating the nitrilase dimers into the oligomer. Here we used cysteine scanning across these two regions, in the spiral forming nitrilase cyanide dihydratase from Bacillus pumilus (CynD), to identify residues altering the oligomeric state or activity of the nitrilase. Several mutations were found to cause changes to the size of the oligomer as well as reduction in activity. Additionally one mutation, R67C, caused a partial defect in oligomerization with the accumulation of smaller oligomer variants. These results support the hypothesis that these insertion regions contribute to the unique quaternary structure of the spiral microbial nitrilases. PMID:26779137

  20. Improving the thermoactivity and thermostability of pectate lyase from Bacillus pumilus for ramie degumming.

    PubMed

    Liang, Chaoning; Gui, Xiwu; Zhou, Cheng; Xue, Yanfen; Ma, Yanhe; Tang, Shuang-Yan

    2015-03-01

    Thermostable alkaline pectate lyases can be potentially used for enzymatically degumming ramie in an environmentally sustainable manner and as an alternative to the currently used chemical-based ramie degumming processes. To assess its potential applications, pectate lyase from Bacillus pumilus (ATCC 7061) was cloned and expressed in Escherichia coli. Evolutionary strategies were applied to generate efficient ramie degumming enzymes. Obtained from site-saturation mutagenesis and random mutagenesis, the best performing mutant enzyme M3 exhibited a 3.4-fold higher specific activity on substrate polygalacturonic acid, compared with the wild-type enzyme. Furthermore, the half-life of inactivation at 50 °C for M3 mutant extended to over 13 h. In contrast, the wild-type enzyme was completely inactivated in less than 10 min under the same conditions. An upward shift in the optimal reaction temperature of M3 mutant, to 75 °C, was observed, which was 10 °C higher than that of the wild-type enzyme. Kinetic parameter data revealed that the catalysis efficiency of M3 mutant was higher than that of the wild-type enzyme. Ramie degumming with M3 mutant was also demonstrated to be more efficient than that with the wild-type enzyme. Collectively, our results suggest that the M3 mutant, with remarkable improvements in thermoactivity and thermostability, has potential applications for ramie degumming in the textile industry. PMID:25287558

  1. Purification and characterization of a soybean-milk-coagulating enzyme from Bacillus pumilus TYO-67.

    PubMed

    Yasuda, M; Aoyama, M; Sakaguchi, M; Nakachi, K; Kobamoto, N

    1999-04-01

    Bacillus pumilus TYO-67 was isolated from tofu (soybean curd) as the best producer of a soybean-milk-coagulating enzyme, induced by the addition of soybean protein to the growth medium. The enzyme was purified approximately 30-fold with an 11% yield. The homogeneous preparation of the enzyme showed that it is a monomer with a molecular mass of about 30 kDa and has an isoelectric point at pH 9.75. The results of amino acid composition analyses showed that the enzyme is rich in alanine, aspartic acid, glycine, serine and valine. Although the amino-terminal amino acid (alanine) was identical with that of subtilisins, the amino-terminal sequence was different from those of subtilisins. The alpha-helix content of the enzyme was calculated to be 28.2%. The optimum pH and temperature were observed at 6.0-6.1 and 65 degrees C respectively. The enzyme was significantly activated by the addition of 1 mM Mn2+, Ca2+, Mg2+, and Sr2+ ions in the reaction mixture, and its thermal stability was significantly increased by Ca2+ ion. PMID:10341429

  2. Site-directed mutagenesis at aspartate and glutamate residues of xylanase from Bacillus pumilus.

    PubMed

    Ko, E P; Akatsuka, H; Moriyama, H; Shinmyo, A; Hata, Y; Katsube, Y; Urabe, I; Okada, H

    1992-11-15

    To elucidate the reaction mechanism of xylanase, the identification of amino acids essential for its catalysis is of importance. Studies have indicated the possibility that the reaction mechanism of xylanase is similar to that of hen's egg lysozyme, which involves acidic amino acid residues. On the basis of this assumption, together with the three-dimensional structure of Bacillus pumilus xylanase and its amino acid sequence similarity to other xylanases of different origins, three acidic amino acids, namely Asp-21, Glu-93 and Glu-182, were selected for site-directed mutagenesis. The Asp residue was altered to either Ser or Glu, and the Glu residues to Ser or Asp. The purified mutant xylanases D21E, D21S, E93D, E93S, E182D and E182S showed single protein bands of about 26 kDa on SDS/PAGE. C.d. spectra of these mutant enzymes show no effect on the secondary structure of xylanase, except that of D21E, which shows a little variation. Furthermore, mutations of Glu-93 and Glu-182 resulted in a drastic decrease in the specific activity of xylanase as compared with mutation of Asp-21. On the basis of these results we propose that Glu-93 and Glu-182 are the best candidates for the essential catalytic residues of xylanase. PMID:1359880

  3. Substrate specificity and thermostability of the dehairing alkaline protease from Bacillus pumilus.

    PubMed

    Wan, Min-Yuan; Wang, Hai-Yan; Zhang, Yi-Zheng; Feng, Hong

    2009-11-01

    An alkaline protease (DHAP) from Bacillus pumilus has shown great potential in hide dehairing. To get better insights on its catalytic properties for application, the substrate specificity and thermostability were investigated using five natural proteins and nine synthetic peptides. The results showed that DHAP could hydrolyze five proteins tested here in different specificity. Collagen, a component of animal skin, was more resistant to hydrolysis than casein, fibrin, and gelatin. Among the synthetic peptides, the enzyme showed activity mainly with tetrapeptide substrates with the catalytic efficiency in order of Phe>Leu>Ala at P1 site, although k(m) value for AAVA-pN is much lower than that for AAPL-pN and AAPF-pN. With tripeptide substrates, smaller side-chain group (Gly) at P1 site was not hydrolyzed by DHAP. The enzyme showed good thermostability below 60 degrees C, and lost activity so quickly above 70 degrees C. The thermostability was largely dependent on metal ion, especially Ca(2+), although other ions, like Mg(2+), Mn(2+), and Co(2+), could sustain stability at certain extent within limited time. Cu(2+), Fe(2+), as well as Al(3+), did not support the enzyme to retain activity at 60 degrees C even in 5 min. In addition, the selected metal ions could coordinate calcium in improvement or destruction of thermostability for DHAP. PMID:19132554

  4. Structural analysis of Bacillus pumilus phenolic acid decarboxylase, a lipocalin-fold enzyme

    SciTech Connect

    Matte, Allan; Grosse, Stephan; Bergeron, Hélène; Abokitse, Kofi; Lau, Peter C.K.

    2012-04-30

    The decarboxylation of phenolic acids, including ferulic and p-coumaric acids, to their corresponding vinyl derivatives is of importance in the flavoring and polymer industries. Here, the crystal structure of phenolic acid decarboxylase (PAD) from Bacillus pumilus strain UI-670 is reported. The enzyme is a 161-residue polypeptide that forms dimers both in the crystal and in solution. The structure of PAD as determined by X-ray crystallography revealed a -barrel structure and two -helices, with a cleft formed at one edge of the barrel. The PAD structure resembles those of the lipocalin-fold proteins, which often bind hydrophobic ligands. Superposition of structurally related proteins bound to their cognate ligands shows that they and PAD bind their ligands in a conserved location within the -barrel. Analysis of the residue-conservation pattern for PAD-related sequences mapped onto the PAD structure reveals that the conservation mainly includes residues found within the hydrophobic core of the protein, defining a common lipocalin-like fold for this enzyme family. A narrow cleft containing several conserved amino acids was observed as a structural feature and a potential ligand-binding site.

  5. Ethanol/Water Pulps From Sugar Cane Straw and Their Biobleaching With Xylanase from Bacillus pumilus

    NASA Astrophysics Data System (ADS)

    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.

  6. Probing an Interfacial Surface in the Cyanide Dihydratase from Bacillus pumilus, A Spiral Forming Nitrilase.

    PubMed

    Park, Jason M; Mulelu, Andani; Sewell, B Trevor; Benedik, Michael J

    2015-01-01

    Nitrilases are of significant interest both due to their potential for industrial production of valuable products as well as degradation of hazardous nitrile-containing wastes. All known functional members of the nitrilase superfamily have an underlying dimer structure. The true nitrilases expand upon this basic dimer and form large spiral or helical homo-oligomers. The formation of this larger structure is linked to both the activity and substrate specificity of these nitrilases. The sequences of the spiral nitrilases differ from the non-spiral forming homologs by the presence of two insertion regions. Homology modeling suggests that these regions are responsible for associating the nitrilase dimers into the oligomer. Here we used cysteine scanning across these two regions, in the spiral forming nitrilase cyanide dihydratase from Bacillus pumilus (CynD), to identify residues altering the oligomeric state or activity of the nitrilase. Several mutations were found to cause changes to the size of the oligomer as well as reduction in activity. Additionally one mutation, R67C, caused a partial defect in oligomerization with the accumulation of smaller oligomer variants. These results support the hypothesis that these insertion regions contribute to the unique quaternary structure of the spiral microbial nitrilases. PMID:26779137

  7. Purification and Characterization of a Fibrinolytic Enzyme from Bacillus pumilus 2.g Isolated from Gembus, an Indonesian Fermented Food

    PubMed Central

    Afifah, Diana Nur; Sulchan, Muhammad; Syah, Dahrul; Yanti; Suhartono, Maggy Thenawidjaja; Kim, Jeong Hwan

    2014-01-01

    Bacillus pumilus 2.g isolated from gembus, an Indonesian fermented soybean cake, secretes several proteases that have strong fibrinolytic activities. A fibrinolytic enzyme with an apparent molecular weight of 20 kDa was purified from the culture supernatant of B. pumilus 2.g by sequential application of ammonium sulfate precipitation, ion-exchange chromatography, and hydrophobic chromatography. The partially purified enzyme was stable between pH 5 and pH 9 and temperature of less than 60°C. Fibrinolytic activity was increased by 5 mM MgCl2 and 5 mM CaCl2 but inhibited by 1 mM phenylmethylsulfonyl fluoride (PMSF), 1 mM sodium dodecyl sulfate (SDS), and 1 mM ethylenediaminetetraacetic acid (EDTA). The partially purified enzyme quickly degraded the ? and ? chains of fibrinogen but was unable to degrade the ? chain. PMID:25320719

  8. Purification and Characterization of a Fibrinolytic Enzyme from Bacillus pumilus 2.g Isolated from Gembus, an Indonesian Fermented Food.

    PubMed

    Afifah, Diana Nur; Sulchan, Muhammad; Syah, Dahrul; Yanti; Suhartono, Maggy Thenawidjaja; Kim, Jeong Hwan

    2014-09-01

    Bacillus pumilus 2.g isolated from gembus, an Indonesian fermented soybean cake, secretes several proteases that have strong fibrinolytic activities. A fibrinolytic enzyme with an apparent molecular weight of 20 kDa was purified from the culture supernatant of B. pumilus 2.g by sequential application of ammonium sulfate precipitation, ion-exchange chromatography, and hydrophobic chromatography. The partially purified enzyme was stable between pH 5 and pH 9 and temperature of less than 60°C. Fibrinolytic activity was increased by 5 mM MgCl2 and 5 mM CaCl2 but inhibited by 1 mM phenylmethylsulfonyl fluoride (PMSF), 1 mM sodium dodecyl sulfate (SDS), and 1 mM ethylenediaminetetraacetic acid (EDTA). The partially purified enzyme quickly degraded the ? and ? chains of fibrinogen but was unable to degrade the ? chain. PMID:25320719

  9. Genotyping of starter cultures of Bacillus subtilis and Bacillus pumilus for fermentation of African locust bean (Parkia biglobosa) to produce Soumbala.

    PubMed

    Ouoba, Labia Irène Ivette; Diawara, Bréhima; Amoa-Awua, Wisdom kofi; Traoré, Alfred Sababénedyo; Møller, Peter Lange

    2004-01-15

    Bacillus spp. are the predominant microorganisms in fermented African locust bean called Soumbala in Burkina Faso. Ten strains selected as potential starter cultures were characterised by PCR amplification of the16S-23S rDNA intergenic transcribed spacer (ITS-PCR), restriction fragment length polymorphism of the ITS-PCR (ITS-PCR RFLP), pulsed field gel electrophoresis (PFGE) and sequencing of the 968-1401 region of the 16S rDNA. In previous studies, the isolates were identified by phenotyping as Bacillus subtilis and Bacillus pumilus. The phenotyping was repeated as a reference in the present study. The ITS-PCR and ITS-PCR RLFP allowed a typing at species level. The PFGE was more discriminative and allowed a typing at strain level. Full agreement with the phenotyping was observed in all cases. The sequencing of the 16S rDNA allowed the identification at species level with an identity from 97% to 100% comparing the sequences to those from the GenBank databases. The desired cultures of B. subtilis and B. pumilus from African locust bean fermentation were distinguished by ITS-PCR and ITS-PCR RLFP from Bacillus cereus and Bacillus sphaericus which sometimes occur in the beginning of the fermentation. PMID:14698101

  10. Spore UV and acceleration resistance of endolithic Bacillus pumilus and Bacillus subtilis isolates obtained from Sonoran desert basalt: implications for lithopanspermia.

    PubMed

    Benardini, James N; Sawyer, John; Venkateswaran, Kasthuri; Nicholson, Wayne L

    2003-01-01

    Bacterial spores have been used as model systems for studying the theory of interplanetary transport of life by natural processes such as asteroidal or cometary impacts (i.e., lithopanspermia). Because current spallation theory predicts that near-surface rocks are ideal candidates for planetary ejection and surface basalts are widely distributed throughout the rocky planets, we isolated spore-forming bacteria from the interior of near-subsurface basalt rocks collected in the Sonoran desert near Tucson, Arizona. Spores were found to inhabit basalt at very low concentrations (Bacillus pumilus and one Bacillus subtilis isolate were recovered from near-subsurface basalt samples. Populations of purified spores prepared from the isolated strains were subjected to 254-nm UV and ballistics tests in order to assess their resistance to UV radiation and to extreme acceleration shock, two proposed lethal factors for spores during interplanetary transfer. Specific natural isolates of B. pumilus were found to be substantially more resistant to UV and extreme acceleration than were reference laboratory strains of B. subtilis, the benchmark organism, suggesting that spores of environmental B. pumilus isolates may be more likely to survive the rigors of interplanetary transfer. PMID:14987476

  11. Spore UV and Acceleration Resistance of Endolithic Bacillus pumilus and Bacillus subtilis Isolates Obtained from Sonoran Desert Basalt: Implications for Lithopanspermia

    NASA Astrophysics Data System (ADS)

    Benardini, James N.; Sawyer, John; Venkateswaran, Kasthuri; Nicholson, Wayne L.

    2003-12-01

    Bacterial spores have been used as model systems for studying the theory of interplanetary transport of life by natural processes such as asteroidal or cometary impacts (i.e., lithopanspermia). Because current spallation theory predicts that near-surface rocks are ideal candidates for planetary ejection and surface basalts are widely distributed throughout the rocky planets, we isolated spore-forming bacteria from the interior of near-subsurface basalt rocks collected in the Sonoran desert near Tucson, Arizona. Spores were found to inhabit basalt at very low concentrations (<=28 colony-forming units/g) in these samples. Six isolates identified as being most closely related to Bacillus pumilus and one Bacillus subtilis isolate were recovered from near-subsurface basalt samples. Populations of purified spores prepared from the isolated strains were subjected to 254-nm UV and ballistics tests in order to assess their resistance to UV radiation and to extreme acceleration shock, two proposed lethal factors for spores during interplanetary transfer. Specific natural isolates of B. pumilus were found to be substantially more resistant to UV and extreme acceleration than were reference laboratory strains of B. subtilis, the benchmark organism, suggesting that spores of environmental B. pumilus isolates may be more likely to survive the rigors of interplanetary transfer.

  12. Phenotypic subtyping and colonization studies of the maize endophyte Bacillus mojavensis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Bacillus mojavensis was discovered in corn kernels and later determined to possess endophytic characteristics. The bacterium was also determined to have biocontrol potential due to its growth inhibition of the maize mycotoxic fungus Fusarium verticillioides. Maize plants with endophytic bacterium al...

  13. Thermostable Xanthine Oxidase Activity from Bacillus pumilus RL-2d Isolated from Manikaran Thermal Spring: Production and Characterization.

    PubMed

    Sharma, Nirmal Kant; Thakur, Shikha; Thakur, Neerja; Savitri; Bhalla, Tek Chand

    2016-03-01

    Xanthine oxidase is an important enzyme of purine metabolism that catalyzes the hydroxylation of hypoxanthine to xanthine and then xanthine to uric acid. A thermostable xanthine oxidase is being reported from a thermophilic organism RL-2d isolated from the Manikaran (Kullu) hot spring of Himachal Pradesh (India). Based on the morphology, physiological tests, and 16S rDNA gene sequence, RL-2d was identified as Bacillus pumilus. Optimization of physiochemical parameters resulted into 4.1-fold increase in the xanthine oxidase activity from 0.051 U/mg dcw (dry cell weight) to 0.209 U/mg dcw. The xanthine oxidase of B. pumilus RL-2d has exhibited very good thermostability and its t1/2 at 70 and 80 °C were 5 and 1 h, respectively. Activity of this enzyme was strongly inhibited by Hg(2+), Ag(+) and allopurinol. The investigation showed that B. pumilus RL-2d exhibited highest xanthine oxidase activity and remarkable thermostability among the other xanthine oxidases reported so far. PMID:26843701

  14. Biochemical and molecular characterization of a detergent-stable serine alkaline protease from Bacillus pumilus CBS with high catalytic efficiency.

    PubMed

    Jaouadi, Bassem; Ellouz-Chaabouni, Semia; Rhimi, Moez; Bejar, Samir

    2008-09-01

    We have described previously the potential use of an alkaline protease from Bacillus pumilus CBS as an effective additive in laundry detergent formulations [B. Jaouadi, S. Ellouz-Chaabouni, M. Ben Ali, E. Ben Messaoud, B. Naili, A. Dhouib, S. Bejar, A novel alkaline protease from Bacillus pumilus CBS having a high compatibility with laundry detergent and a high feather-degrading activity, Process Biochem, submitted for publication]. Here, we purified this enzyme (named SAPB) and we cloned, sequenced and over-expressed the corresponding gene. The enzyme was purified to homogeneity using salt precipitation and gel filtration HPLC. The pure protease was found to be monomeric protein with a molecular mass of 34598.19Da as determined by MALDI-TOF mass spectrometry. The NH2-terminal sequence of first 21 amino acids (aa) of the purified SAPB was AQTVPYGIPQIKAPAVHAQGY and was completely identical to proteases from other Bacillus pumilus species. This protease is strongly inhibited by PMSF and DFP, showing that it belongs to the serine proteases superfamily. Interestingly, the optimum pH is 10.6 while the optimum temperature was determined to be 65 degrees C. The enzyme was completely stable within a wide range of pH (7.0-10.6) and temperature (30-55 degrees C). One of the distinguishing properties is its catalytic efficiency (kcat/Km) calculated to be 45,265min(-1)mM(-1) and 147,000min(-1)mM(-1) using casein and AAPF as substrates, respectively, which is higher than that of Subtilisin Carlsberg, Subtilisin BPN' and Subtilisin 309 determined under the same conditions. In addition, SAPB showed remarkable stability, for 24h at 40 degrees C, in the presence of 5% Tween-80, 1% SDS, 15% urea and 10% H2O2, which comprise the common bleach-based detergent formulation. The sapB gene encoding SAPB was cloned, sequenced and over-expressed in Escherichia coli. The purified recombinant enzyme (rSAPB) has the same physicochemical and kinetic properties as the native one. SapB gene had an ORF of 1149bp encoding a protein of 383 aa organized into a signal peptide (29 aa), a pro-protein (79 aa) and a mature enzyme (275 aa). The deduced amino acid sequence inspection displays an important homology with other bacterial proteases. The highest homology of 98.1% was found with BPP-A protease from Bacillus pumilus MS-1, with only 8 aa of difference. PMID:18397761

  15. Purification and properties of an polygalacturonic acid trans-eliminase produced by Bacillus pumilus.

    PubMed

    Davé, B A; Vaughn, R H

    1971-10-01

    A strain of Bacillus pumilus produced an extracellular pectic enzyme with polygalacturonic acid as the substrate. This enzyme, with optimal activity at pH 8.0 to 8.5, produced reaction products that strongly absorbed light at 232 nm, indicating the presence of a pectic acid trans-eliminase (PATE). Neither pectin esterase nor polygalacturonase was detected in the cell-free culture fluid. Chromatographic examination of the end products revealed the presence of large quantities of unsaturated oligouronides unlike those found with B. polymyxa. It was found that the PATE was produced extracellularly during the negative logarithmic death phase of the organism. The filtrate from sonically treated cells did not show any activity for PATE or hydrolases for lower oligogalacturonides at any time during the growth cycle. The enzyme was inducible. Pectin, National Formulary (NF) was the best inducer, followed by polygalacturonic acid and galacturonic acid. Enzyme activity was markedly stimulated by calcium and other divalent ions. Copper and cobalt ions were inhibitory. The partially purified enzyme showed no significant activity on pectin containing a high methoxyl content (96% esterified). However, pectin NF with a lower methoxyl content (68% esterified) was attacked to a degree by the partially purified and crude enzyme preparations. The initial rate of PATE activity increased up to 60 C, about 16-fold higher than that observed at room temperature. The activation energy was calculated as 12,183 cal/mole. A protective action of calcium chloride against heat inactivation of the PATE was observed. Degradation of polygalacturonic acid by this enzyme produced several unsaturated oligouronides soon after its addition to the substrate. The major endproduct was thought to be different from that of other known PATE enzymes. Paper chromatographic studies and viscosity measurements disclosed the random cleaving nature of the enzyme an endo-PATE. PMID:5122802

  16. CotA, a Multicopper Oxidase from Bacillus pumilus WH4, Exhibits Manganese-Oxidase Activity

    PubMed Central

    Su, Jianmei; Bao, Peng; Bai, Tenglong; Deng, Lin; Wu, Hui; Liu, Fan; He, Jin

    2013-01-01

    Multicopper oxidases (MCOs) are a family of enzymes that use copper ions as cofactors to oxidize various substrates. Previous research has demonstrated that several MCOs such as MnxG, MofA and MoxA can act as putative Mn(II) oxidases. Meanwhile, the endospore coat protein CotA from Bacillus species has been confirmed as a typical MCO. To study the relationship between CotA and the Mn(II) oxidation, the cotA gene from a highly active Mn(II)-oxidizing strain Bacillus pumilus WH4 was cloned and overexpressed in Escherichia coli strain M15. The purified CotA contained approximately four copper atoms per molecule and showed spectroscopic properties typical of blue copper oxidases. Importantly, apart from the laccase activities, the CotA also displayed substantial Mn(II)-oxidase activities both in liquid culture system and native polyacrylamide gel electrophoresis. The optimum Mn(II) oxidase activity was obtained at 53°C in HEPES buffer (pH 8.0) supplemented with 0.8 mM CuCl2. Besides, the addition of o-phenanthroline and EDTA both led to a complete suppression of Mn(II)-oxidizing activity. The specific activity of purified CotA towards Mn(II) was 0.27 U/mg. The Km, Vmax and kcat values towards Mn(II) were 14.85±1.17 mM, 3.01×10?6±0.21 M·min?1 and 0.32±0.02 s?1, respectively. Moreover, the Mn(II)-oxidizing activity of the recombinant E. coli strain M15-pQE-cotA was significantly increased when cultured both in Mn-containing K liquid medium and on agar plates. After 7-day liquid cultivation, M15-pQE-cotA resulted in 18.2% removal of Mn(II) from the medium. Furthermore, the biogenic Mn oxides were clearly observed on the cell surfaces of M15-pQE-cotA by scanning electron microscopy. To our knowledge, this is the first report that provides the direct observation of Mn(II) oxidation with the heterologously expressed protein CotA, Therefore, this novel finding not only establishes the foundation for in-depth study of Mn(II) oxidation mechanisms, but also offers a potential biocatalyst for Mn(II) removal. PMID:23577125

  17. Candidate Genes That May Be Responsible for the Unusual Resistances Exhibited by Bacillus pumilus SAFR-032 Spores

    PubMed Central

    Tirumalai, Madhan R.; Rastogi, Rajat; Zamani, Nader; O’Bryant Williams, Elisha; Allen, Shamail; Diouf, Fatma; Kwende, Sharon; Weinstock, George M.; Venkateswaran, Kasthuri J.; Fox, George E.

    2013-01-01

    The spores of several Bacillus species, including Bacillus pumilus SAFR-032 and B. safensis FO-36b, which were isolated from the spacecraft assembly facility at NASA's Jet Propulsion Laboratory, are unusually resistant to UV radiation and hydrogen peroxide. In order to identify candidate genes that might be associated with these resistances, the whole genome of B. pumilus SAFR-032, and the draft genome of B. safensis FO-36b were compared in detail with the very closely related type strain B. pumilus ATCC7061T. 170 genes are considered characteristic of SAFR-032, because they are absent from both FO-36b and ATCC7061T. Forty of these SAFR-032 characteristic genes are entirely unique open reading frames. In addition, four genes are unique to the genomes of the resistant SAFR-032 and FO-36b. Fifty three genes involved in spore coat formation, regulation and germination, DNA repair, and peroxide resistance, are missing from all three genomes. The vast majority of these are cleanly deleted from their usual genomic context without any obvious replacement. Several DNA repair and peroxide resistance genes earlier reported to be unique to SAFR-032 are in fact shared with ATCC7061T and no longer considered to be promising candidates for association with the elevated resistances. Instead, several SAFR-032 characteristic genes were identified, which along with one or more of the unique SAFR-032 genes may be responsible for the elevated resistances. These new candidates include five genes associated with DNA repair, namely, BPUM_0608 a helicase, BPUM_0652 an ATP binding protein, BPUM_0653 an endonuclease, BPUM_0656 a DNA cytosine-5- methyltransferase, and BPUM_3674 a DNA helicase. Three of these candidate genes are in immediate proximity of two conserved hypothetical proteins, BPUM_0654 and BPUM_0655 that are also absent from both FO-36b and ATCC7061T. This cluster of five genes is considered to be an especially promising target for future experimental work. PMID:23799069

  18. Applicability of UV resistant Bacillus pumilus endospores as a human adenovirus surrogate for evaluating the effectiveness of virus inactivation in low-pressure UV treatment systems

    EPA Science Inventory

    Recent studies have demonstrated the potential to use Bacillus pumilus endospores as a surrogate of human adenovirus (HAdV) in UV disinfection studies. The use of endospores has been limited by observations of batch-to-batch variation in UV sensitivity. This study reports on a pr...

  19. Bacillus pumilus of Palk Bay origin inhibits quorum-sensing-mediated virulence factors in Gram-negative bacteria.

    PubMed

    Nithya, Chari; Aravindraja, Chairman; Pandian, Shunmugiah Karutha

    2010-05-01

    The aim of the current study was to inhibit quoring-sensing(QS)-mediated virulence factors of representative Gram-negative bacteria by marine bacterial isolates. Bacteria isolated from Palk Bay sediments were screened for anti-QS activity. Eleven strains inhibited QS signals in Chromobacterium violaceum (ATCC 12472) and C. violaceum CV026. The marine bacterial strain S8-07 reduced the accumulation of N-acyl homoserine lactone (AHLs) and showed significant inhibition of LasA protease(76%), LasB elastase(84%), caseinase(70%), pyocyanin (84%), pyoverdin and biofilm formation(87%) in Pseudomonas aeruginosa PAO1. Strain S8-07 also showed highly significant reduction (90%) in prodigiosin, secreted casienase (92%), hemolytic activity (73%) and biofilm formation (61%) in Serratia marcescens. Strain S8-07, identified as Bacillus pumilus (accession number FJ584416), showed distinct profiles of inhibition against the virulence factors of both P. aeruginosa PAO1 (las, rhl) and S. marcescens (shl). Polar extraction and proteinase K treatment of the culture supernatant confirmed that the anti-QS activity of S8-07 was indeed due to a protein molecule. Acidification assay and HPLC analysis revealed that the degradation of AHL was not due to lactonase activity, but rather, was due to acylase activity of S8-07. Thus, novel anti-QS acylase activity is reported for the first time from a B. pumilus strain of marine origin. PMID:20381609

  20. Phylogenetic and clonality analysis of Bacillus pumilus isolates uncovered a highly heterogeneous population of different closely related species and clones.

    PubMed

    Branquinho, Raquel; Meirinhos-Soares, Luís; Carriço, João A; Pintado, Manuela; Peixe, Luísa V

    2014-12-01

    Bacillus pumilus is a Gram-positive bacterium with a wide range of attributed applications, namely as a plant growth promoting rhizobacteria (PGPR), animal, and human probiotic. However, a rare putative role in human diseases has been reported, namely in food poisoning or as anthrax-like cutaneous infectious agent. This species is difficult to distinguish from its closely related species on the basis of phenotypic or biochemical characteristics and 16S rRNA gene sequences. In this study, the phylogenetic analysis of gyrB and rpoB gene sequences of a collection of isolates previously identified as B. pumilus, assigned most of them (93%, 38 of 41 isolates) to B. safensis or to the new recently described B. invictae. Moreover, we extended the previously reported recognized habitats of these species and unveiled a human health or biotechnological relevance (e.g. as implicated in food poisoning or PGPR) for them. Additionally, we demonstrated that both B. safensis and B. invictae species encompass a clonally diverse population, which can justify their great adaptation ability to different niches, with evidence of clonal-host specificity. PMID:25230950

  1. Production of an alkaline protease using Bacillus pumilus D3 without inactivation by SDS, its characterization and purification.

    PubMed

    Özçelik, Burçin; Aytar, P?nar; Gedikli, Serap; Yard?mc?, Ezgi; Çal??kan, Figen; Çabuk, Ahmet

    2014-06-01

    Abstract In this study, protease-producing capacity of Bacillus pumilus D3, isolated from hydrocarbon contaminated soil, was evaluated and optimized. Optimum growing conditions for B. pumilus D3 in terms of protease production were determined as 1% optimum inoculum size, 35?°C temperature, 11 pH and 48?h incubation time, respectively. Stability studies indicated that the mentioned protease was stable within the pH range of 7-10.5 and between 30?°C and 40?°C temperatures. Surprisingly, the activity of the enzyme increased in the presence of SDS with concentration up to 5?mM. The protease was concentrated 1.6-fold with ammonium sulfate precipitation and dialysis. At least six protein bands were obtained from dialysate by electrophoresis. Four clear protein bands with caseinolytic activity were detected by zymography. Dialysate was further purified by anion-exchange chromatography and the caseinolytic active fraction showed a single band between 29 and 36?kDa of reducing conditions. PMID:23638694

  2. An ICEBs1-like element may be associated with the extreme radiation and desiccation resistance of Bacillus pumilus SAFR-032 spores.

    PubMed

    Tirumalai, Madhan R; Fox, George E

    2013-09-01

    Comparisons of the genomes of Bacillus pumilus SAFR-032 and the closely related type strain, B. pumilus ATCC7061(T), exposed an extended region of non-homologous genes. A detailed examination of this region revealed the presence of an ICEBs1-like integrative conjugative element in SAFR-032. A similar element was subsequently located elsewhere in the ATCC7061(T) genome. A detailed comparison of these elements and the ICEBs1 of B. subtilis revealed extremely rapid flux in gene content, genome organization and sequence similarity. It is not clear if the B. pumilus elements as they are currently structured are functional. However, it is clear that the past involvement of these elements has brought multiple genes of unknown function to the SAFR-032 genome and these genes may be responsible for the rapid evolution that led to the extreme radiation and desiccation resistance of this organism's spores. PMID:23812891

  3. Enhanced phytotransformation of Navy Blue RX dye by Petunia grandiflora Juss. with augmentation of rhizospheric Bacillus pumilus strain PgJ and subsequent toxicity analysis.

    PubMed

    Watharkar, Anuprita D; Rane, Niraj R; Patil, Swapnil M; Khandare, Rahul V; Jadhav, Jyoti P

    2013-08-01

    This study reveals the beneficial synergistic phytoremediation potential of Petunia grandiflora Juss. with its rhizospheric bacterial isolate Bacillus pumilus strain PgJ to decolorize reactive Navy Blue RX (NBRX) dye by their active enzymatic machinery. In vitro cultures of P. grandiflora and B. pumilus gave 80.01% and 76.80% while their consortium decolorized NBRX up to 96.86% within 36 h. Significant induction in the enzyme activities of lignin peroxidase (207%), tyrosinase (133%), laccase (161%), riboflavin reductase (78%) were seen in the roots of tissue cultured plants while enzymes tyrosinase (660%), laccase (689%), riboflavin reductase (528%) were induced significantly in the B. pumilus cells. Metabolites of treated NBRX were analyzed using UV-vis spectroscopy, gas chromatography and biotransformation was visualized using high performance thin layer chromatography profile. Metabolites of the dye exhibited reduced phytotoxicity Sorghum vulgare and Phaeseolus mungo and significant reduction in cytogenotoxicity on Allium cepa roots when compared to NBRX. PMID:23743429

  4. Characterization of industrially-valuable xylano-pectinolytic enzymes produced concurrently by a novel isolate of Bacillus pumilus.

    PubMed

    Kaur, Amanjot; Singh, Avtar; Mahajan, Ritu

    2014-11-01

    Xylanase and polygalacturonase were concurrently produced by a novel alkalo-thermotolerant Bacillus pumilus AJK. They were purified and characterized to evaluate their potential for various industrial applications. Xylanase was purified to 19-fold with 67 % recovery and polygalacturonase up to 23-fold with 75 % recovery. Existence of multiple forms of xylanase was indicated by its elution-profile through Sephadex G-100 as two peaks, xylanase-I and xylanase-II, with molecular weights of ~24.5 and ~13 kDa, respectively, and by the presence of two pH optima, one at pH 6.0 and other at pH 8.5. The molecular weight of polygalacturonase was ~40 kDa by gel-filtration chromatography. Zymographic studies confirmed the presence of seven isozymic forms of xylanase. Xylanase and polygalacturonase are stable over a broad range of pH and temperature. PMID:25048224

  5. An alkali-thermostable xylanase from Bacillus pumilus functionally expressed in Kluyveromyces lactis and evaluation of its deinking efficiency.

    PubMed

    Thomas, Leya; Ushasree, Mrudula V; Pandey, Ashok

    2014-08-01

    This work aimed at studying the recombinant expression of an alkali- and thermo-stable xylanase from Bacillus pumilus in Kluyveromyces lactis and its use in deinking of civic paper waste. Efficient expression with a 3-fold increase in the activity than the native organism was achieved. An inducer concentration of 2.5% and medium pH of 9.0 was the best for enzyme expression. Purified enzyme showed an optimum activity at temperatures 50 and 60°C and pH 9.0 and 10.0, respectively. At pH 12.0, enzyme retained 74% and 26% activity after 2 and 3h of incubation, respectively. After incubation at 50 and 60°C for 1h, the enzyme showed 100% retention of activity, and remained active for 4h at 60°C retaining 23% residual activity. Partially purified recombinant enzyme showed higher deinking efficiency (273%) of laser print waste paper than crude xylanase from Bacillus and commercial acidic enzyme. This xylanase with superior stability characteristics could be a suitable candidate in paper and pulp industries. PMID:24709528

  6. Immobilization of xylanase purified from Bacillus pumilus VLK-1 and its application in enrichment of orange and grape juices.

    PubMed

    Kumar, Lalit; Nagar, Sushil; Mittal, Anuradha; Garg, Neelam; Gupta, Vijay Kumar

    2014-09-01

    This study was conducted to evaluate the efficacy of purified free and immobilized xylanase in enrichment of fruit juices. Extracellular xylanase produced from Bacillus pumilus VLK-1 was purified to apparent homogeneity by 15.4-fold with 88.3 % recovery in a single step using CM-Sephadex C-50. Purified xylanase showed a single band on SDS-polyacrylamide gel with a molecular mass of 22.0 kDa. The purified enzyme was immobilized on glutaraldehyde-activated aluminum oxide pellets and the immobilization process parameters were optimized statistically through response surface methodology. The bound enzyme displayed an increase in optimum temperature from 60 to 65?ºC and pH from 8.0 to 9.0. The pH and temperature stability of the enzyme was also enhanced after immobilization. It could be reused for 10 consecutive cycles with 58 % residual enzyme activity. The potential of purified xylanase (free and immobilized) in juice enrichment from grape (Vitis amurensis) and orange (Citrus sinensis) pulps has been investigated. The optimization of this process using free xylanase revealed maximum juice yield, clarity and reducing sugar on treatment with 20 IU/g fruit pulp for 30 min at 50?ºC. Treatment of both the fruit pulps with xylanase under optimized conditions resulted in an increase in juice yield, clarity, reducing sugars, titratable acidity, and filterability but a decline in turbidity and viscosity. Immobilized enzyme was more effective in improving juice quality as compared to its soluble counterpart. The results showed B. pumilus VLK-1 xylanase, in both free and immobilized form, as a potential candidate for use in fruit juice enrichment. PMID:25190829

  7. TECHNIQUES FOR MANIPULATING THE BACTERIAL ENDOPHYTE BACILLUS MOJAVENSIS

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The importance of Fusarium species as symptomless endophytes in plant roots is reviewed. Fusarium is a very important genus from the aspects of food production and food safety and its species exist as intercellular root endophytes in both cultivated and wild plants and the role during the symptomle...

  8. High-Quality Draft Genome Sequence of Bacillus amyloliquefaciens Strain 629, an Endophyte from Theobroma cacao

    PubMed Central

    SantAnna, Brena M. M.; Marbach, Phellippe P. A.; Rojas-Herrera, Marcelo; De Souza, Jorge T.; Roque, Milton R. A.

    2015-01-01

    Bacillus amyloliquefaciens strain 629 is an endophyte isolated from Theobroma cacao L. Here, we report the draft genome sequence (3.9 Mb) of B. amyloliquefaciens strain 629 containing 16 contigs (3,903,367 bp), 3,912 coding sequences, and an average 46.5% G+C content. PMID:26586881

  9. High-Quality Draft Genome Sequence of Bacillus amyloliquefaciens Strain 629, an Endophyte from Theobroma cacao.

    PubMed

    SantAnna, Brena M M; Marbach, Phellippe P A; Rojas-Herrera, Marcelo; De Souza, Jorge T; Roque, Milton R A; Queiroz, Artur T L

    2015-01-01

    Bacillus amyloliquefaciens strain 629 is an endophyte isolated from Theobroma cacao L. Here, we report the draft genome sequence (3.9 Mb) of B. amyloliquefaciens strain 629 containing 16 contigs (3,903,367 bp), 3,912 coding sequences, and an average 46.5% G+C content. PMID:26586881

  10. Applicability of UV resistant Bacillus pumilus endospores as a human adenovirus surrogate for evaluating the effectiveness of virus inactivation in low-pressure UV treatment systems.

    PubMed

    Boczek, Laura A; Rhodes, Eric R; Cashdollar, Jennifer L; Ryu, Jongseong; Popovici, Jonathan; Hoelle, Jill M; Sivaganesan, Mano; Hayes, Samuel L; Rodgers, Mark R; Ryu, Hodon

    2016-03-01

    Recent studies have demonstrated the potential to use Bacillus pumilus endospores as a surrogate for human adenovirus (HAdV) in UV disinfection studies. The use of endospores has been limited by observations of batch-to-batch variation in UV sensitivity. This study reports on a propagation method that utilizes a commercially available medium to produce UV tolerant B. pumilus endospores with a consistent UV sensitivity. It is further demonstrated that the endospores of B. pumilus strain (ATCC 27142), produced using this protocol (half strength Columbia broth, 5days incubation, with 0.1mM MnSO4), display a UV dose-response that is similar to that of HAdV. Endospore stocks could be stored in ethanol for up to two months at 4°C without a significant change in UV sensitivity. Synergistic endospore damage was observed by pre-heat treatment of water samples followed by UV irradiation. UV tolerant B. pumilus endospores are a potential surrogate of HAdV for UV treatment performance tests in water utilities which do not have in-house research virology laboratories. PMID:26825005

  11. Structural Characterization and Antimicrobial Activity of a Biosurfactant Obtained From Bacillus pumilus DSVP18 Grown on Potato Peels

    PubMed Central

    Sharma, Deepak; Ansari, Mohammad Javed; Gupta, Sonam; Al Ghamdi, Ahmad; Pruthi, Parul; Pruthi, Vikas

    2015-01-01

    Background: Biosurfactants constitute a structurally diverse group of surface-active compounds derived from microorganisms. They are widely used industrially in various industrial applications such as pharmaceutical and environmental sectors. Major limiting factor in biosurfactant production is their production cost. Objectives: The aim of this study was to investigate biosurfactant production under laboratory conditions with potato peels as the sole source of carbon source. Materials and Methods: A biosurfactant-producing bacterial strain (Bacillus pumilus DSVP18, NCBI GenBank accession no. GQ865643) was isolated from motor oil contaminated soil samples. Biochemical characteristics of the purified biosurfactant were determined and its chemical structure was analyzed. Stability studies were performed and biological activity of the biosurfactant was also evaluated. Results: The strain, when grown on modified minimal salt media supplemented with 2% potato peels as the sole carbon source, showed the ability to reduce Surface Tension (ST) value of the medium from 72 to 28.7 mN/m. The isolated biosurfactant (3.2 ± 0.32 g/L) was stable over a wide range of temperatures (20 - 120 ºC), pH (2-12) and salt concentrations (2 - 12%). When characterized using high-performance liquid chromatography (HPLC) and Fourier transform infrared spectroscopy, it was found to be a lipopeptide in nature, which was further confirmed by Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (mass peak 1044.60) and nuclear magnetic resonance (NMR) studies. Data showed that the isolated biosurfactant at the concentration range of 30 - 35 µg/ml had strong antimicrobial activity when tested against standard strains of Bacillus cereus, Escherichia coli, Salmonella enteritidis, Staphylococcus aureus and Paenibacillus larvae. Conclusions: Potato peels were proved to be potentially useful substrates for biosurfactant production by B. pumilus DSVP18. The strain possessed a unique property to reduce surface tension of the media from 72 to 28.7 mN/m. In addition, it showed a stable surface activity over a wide range of temperatures, pH, and saline conditions and had strong antimicrobial activity. This potential of the identified biosurfactant can be exploited by pharmaceutical industries for its commercial usage. PMID:26495106

  12. Plant genotype-specific archaeal and bacterial endophytes but similar Bacillus antagonists colonize Mediterranean olive trees.

    PubMed

    Müller, Henry; Berg, Christian; Landa, Blanca B; Auerbach, Anna; Moissl-Eichinger, Christine; Berg, Gabriele

    2015-01-01

    Endophytes have an intimate and often symbiotic interaction with their hosts. Less is known about the composition and function of endophytes in trees. In order to evaluate our hypothesis that plant genotype and origin have a strong impact on both, endophytes of leaves from 10 Olea europaea L. cultivars from the Mediterranean basin growing at a single agricultural site in Spain and from nine wild olive trees located in natural habitats in Greece, Cyprus, and on Madeira Island were studied. The composition of the bacterial endophytic communities as revealed by 16S rRNA gene amplicon sequencing and the subsequent PCoA analysis showed a strong correlation to the plant genotypes. The bacterial distribution patterns were congruent with the plant origins in "Eastern" and "Western" areas of the Mediterranean basin. Subsequently, the endophytic microbiome of wild olives was shown to be closely related to those of cultivated olives of the corresponding geographic origins. The olive leaf endosphere harbored mostly Proteobacteria, followed by Firmicutes, Actinobacteria, and Bacteroidetes. The detection of a high portion of archaeal taxa belonging to the phyla Thaumarchaeota, Crenarchaeota, and Euryarchaeota in the amplicon libraries was an unexpected discovery, which was confirmed by quantitative real-time PCR revealing an archaeal portion of up to 35.8%. Although the function of these Archaea for their host plant remains speculative, this finding suggests a significant relevance of archaeal endophytes for plant-microbe interactions. In addition, the antagonistic potential of culturable endophytes was determined; all isolates with antagonistic activity against the olive-pathogenic fungus Verticillium dahliae Kleb. belong to Bacillus amyloliquefaciens. In contrast to the specific global structural diversity, BOX-fingerprints of the antagonistic Bacillus isolates were highly similar and independent of the olive genotype from which they were isolated. PMID:25784898

  13. Plant genotype-specific archaeal and bacterial endophytes but similar Bacillus antagonists colonize Mediterranean olive trees

    PubMed Central

    Müller, Henry; Berg, Christian; Landa, Blanca B.; Auerbach, Anna; Moissl-Eichinger, Christine; Berg, Gabriele

    2015-01-01

    Endophytes have an intimate and often symbiotic interaction with their hosts. Less is known about the composition and function of endophytes in trees. In order to evaluate our hypothesis that plant genotype and origin have a strong impact on both, endophytes of leaves from 10 Olea europaea L. cultivars from the Mediterranean basin growing at a single agricultural site in Spain and from nine wild olive trees located in natural habitats in Greece, Cyprus, and on Madeira Island were studied. The composition of the bacterial endophytic communities as revealed by 16S rRNA gene amplicon sequencing and the subsequent PCoA analysis showed a strong correlation to the plant genotypes. The bacterial distribution patterns were congruent with the plant origins in “Eastern” and “Western” areas of the Mediterranean basin. Subsequently, the endophytic microbiome of wild olives was shown to be closely related to those of cultivated olives of the corresponding geographic origins. The olive leaf endosphere harbored mostly Proteobacteria, followed by Firmicutes, Actinobacteria, and Bacteroidetes. The detection of a high portion of archaeal taxa belonging to the phyla Thaumarchaeota, Crenarchaeota, and Euryarchaeota in the amplicon libraries was an unexpected discovery, which was confirmed by quantitative real-time PCR revealing an archaeal portion of up to 35.8%. Although the function of these Archaea for their host plant remains speculative, this finding suggests a significant relevance of archaeal endophytes for plant–microbe interactions. In addition, the antagonistic potential of culturable endophytes was determined; all isolates with antagonistic activity against the olive-pathogenic fungus Verticillium dahliae Kleb. belong to Bacillus amyloliquefaciens. In contrast to the specific global structural diversity, BOX-fingerprints of the antagonistic Bacillus isolates were highly similar and independent of the olive genotype from which they were isolated. PMID:25784898

  14. Exo-pectinase production by Bacillus pumilus using different agricultural wastes and optimizing of medium components using response surface methodology.

    PubMed

    Tepe, Ozlem; Dursun, Arzu Y

    2014-01-01

    In this research, the production of exo-pectinase by Bacillus pumilus using different agricultural wastes was studied. Agricultural wastes containing pectin such as wheat bran, sugar beet pulp, sunflower plate, orange peel, banana peel, apple pomace and grape pomace were tested as substrates, and activity of exo-pectinase was determined only in the mediums containing sugar beet pulp and wheat bran. Then, effects of parameters such as concentrations of solid substrate (wheat bran and sugar beet pulp) (A), ammonium sulphate (B) and yeast extract (C) on the production of exo-pectinase were investigated by response surface methodology. First, wheat bran was used as solid substrate, and it was determined that exo-pectinase activity increased when relatively low concentrations of ammonium sulphate (0.12-0.21%?w/v) and yeast extract (0.12-0.3%?w/v) and relatively high wheat bran (~5-6%?w/v) were used. Then, exo-pectinase production was optimized by response surface methodology using sugar beet pulp as a solid substrate. In comparison to P values of the coefficients, values of not greater than 0.05 of A and B (2) showed that the effect of these process variables in exo-pectinase production was important and that changes done in these variables will alter the enzyme activity. PMID:24819433

  15. Kinetic studies of Gly28:Ser mutant form of Bacillus pumilus lipase: changes in k(cat) and thermal dependence.

    PubMed

    Bustos-Jaimes, Ismael; Mora-Lugo, Rodrigo; Calcagno, Mario L; Farrés, Amelia

    2010-12-01

    Lipases are useful catalysts for a wide variety of industrial purposes. Herein we report the stability and thermal dependence of the activity of wild-type Bacillus pumilus lipase (BplA) and four site-directed mutants designed to improve its thermal stability. The Gly28:Ser mutation produces a dramatic four-fold increase in its k(cat) and a remarkable increase in its stability. While the increase in k(cat) is temperature-independent, the increase in stability shows that the resultant interactions of this mutation have a strong enthalpic component. Thermal dependence of stability, k(cat), K(M) and k(cat)/K(M) were analysed to gain insight on the structural effects of mutations on BplA. Our results are consistent with a gain in enzyme mobility for those mutants displaying enhanced catalytic properties; the analysis of thermal dependence of kinetic parameters indicates that the mutations did not change either the catalytic mechanism or the rate-limiting step of catalysis. PMID:20831908

  16. Analysis of a conserved hydrophobic pocket important for the thermostability of Bacillus pumilus chloramphenicol acetyltransferase (CAT-86).

    PubMed

    Chirakkal, H; Ford, G C; Moir, A

    2001-03-01

    Site-directed mutagenesis was carried out on Bacillus pumilus chloramphenicol acetyltransferase (CAT-86) to determine the effects of substitution at a conserved hydrophobic pocket identified earlier as important for thermostability. Mutations were introduced that would substitute residues at consensus positions 33, 191 and 203 in the enzyme, both individually and in combination. Two mutants, SDM1 (CAT-86 Y33F, A203V) and SDM5 (CAT-86 A203I), were more thermostable than wild-type and two mutants, SDM4 (CAT-86 I191V) and SDM7 (CAT-86 A203G), were less stable. Reconstruction of the residues of this hydrophobic pocket to that of a more thermostable CAT-R387 enzyme pocket (as a Y33F, I191V, A203V triple mutant) increased the thermostability of the enzyme above the wild-type, but its stability was less than that of SDM1 and SDM5. The K(m) values of the mutant enzymes for chloramphenicol and acetyl-CoA were essentially unaltered (in the ranges 15-30 and 26-35 microM respectively) and the specific activity of purified enzyme was in the range 270-710 units/mg protein. The possible effects of the amino acid substitutions on the CAT-86 structure were determined by homology modelling. A reduction in conformational strain and optimized hydrophobic interactions are predicted to be responsible for the increased thermostability of the SDM1 and SDM5 mutants. PMID:11342712

  17. Identification, sequencing and comparative analysis of pBp15.S plasmid from the newly described entomopathogen Bacillus pumilus 15.1.

    PubMed

    Garcia-Ramon, Diana C; Luque-Navas, Maria Jose; Molina, C Alfonso; Del Val, Coral; Osuna, Antonio; Vilchez, Susana

    2015-11-01

    The Bacillus pumilus 15.1 strain, a recently described entomopathogenic strain active against Ceratitis capitata, contains at least two extrachromosomal elements, pBp15.1S and pBp15.1B. Given that B. pumilus is not a typical entomopathogenic bacterium, the acquisition of this extrachromosomal DNA may explain why B. pumilus 15.1 is toxic to an insect. One of the plasmids present in the strain, the pBp15.1S plasmid, was sub-cloned, sequenced and analyzed using bioinformatics to identify any potential virulence factor. The pBp15.1S plasmid was found to be 7785 bp in size with a GC content of 35.7% and 11 putative ORFs. A replication module typical of a small rolling circle plasmid and a sensing and regulatory system specific for plasmids was found in pBp15.1S. Additionally, we demonstrated the existence of ssDNA in plasmid preparations suggesting that pBp15.1S replicates by the small rolling circle mechanism. A gene cluster present in plasmid pPZZ84 from a distantly isolated B. pumilus strain was also present in pBp15.1S. The plasmid copy number of pBp15.1S in exponentially growing B. pumilus cells was determined to be 33 copies per chromosome. After an extensive plasmid characterization, no known virulence factor was found so a search in the other extrachromosomal elements of the bacteria is needed. PMID:26416357

  18. Endophytic Bacillus spp. produce antifungal lipopeptides and induce host defence gene expression in maize.

    PubMed

    Gond, Surendra K; Bergen, Marshall S; Torres, Mónica S; White, James F

    2015-03-01

    Endophytes are mutualistic symbionts within healthy plant tissues. In this study we isolated Bacillus spp. from seeds of several varieties of maize. Bacillus amyloliquifaciens or Bacillus subtilis were found to be present in all maize varieties examined in this study. To determine whether bacteria may produce antifungal compounds, generally lipopeptides in Bacillus spp., bacterial cultures were screened for production of lipopeptides. Lipopeptides were extracted by acid precipitation from liquid cultures of Bacillus spp. Lipopeptide extracts from Bacillus spp. isolated from Indian popcorn and yellow dent corn showed inhibitory activity against Fusarium moniliforme at 500μg per disk. Using MALDI-TOF mass spectrometry we detected the presence of antifungal iturin A, fengycin and bacillomycin in these isolates. PCR amplification also showed the presence of genes for iturin A and fengycin. B. subtilis (SG_JW.03) isolated from Indian popcorn showed strong inhibition of Arabidopsis seed mycoflora and enhanced seedling growth. We tested for the induction of defence gene expression in the host plant after treatment of plants with B. subtilis (SG_JW.03) and its lipopeptide extract using RT-qPCR. Roots of Indian popcorn seedlings treated with a suspension of B. subtilis (SG_JW.03) showed the induction of pathogenesis-related genes, including PR-1 and PR-4, which relate to plant defence against fungal pathogens. The lipopeptide extract alone did not increase the expression of these pathogenesis-related genes. Based on our study of maize endophytes, we hypothesize that, bacterial endophytes that naturally occur in many maize varieties may function to protect hosts by secreting antifungal lipopeptides that inhibit pathogens as well as inducing the up-regulation of pathogenesis-related genes of host plants (systemic acquired resistance). PMID:25497916

  19. Occurrence of Bacillus amyloliquefaciens as a systemic endophyte of vanilla orchids.

    PubMed

    White, James F; Torres, Mónica S; Sullivan, Raymond F; Jabbour, Rabih E; Chen, Qiang; Tadych, Mariusz; Irizarry, Ivelisse; Bergen, Marshall S; Havkin-Frenkel, Daphna; Belanger, Faith C

    2014-11-01

    We report the occurrence of Bacillus amyloliquefaciens in vanilla orchids (Vanilla phaeantha) and cultivated hybrid vanilla (V. planifolia × V. pompona) as a systemic bacterial endophyte. We determined with light microscopy and isolations that tissues of V. phaeantha and the cultivated hybrid were infected by a bacterial endophyte and that shoot meristems and stomatal areas of stems and leaves were densely colonized. We identified the endophyte as B. amyloliquefaciens using DNA sequence data. Since additional endophyte-free plants and seed of this orchid were not available, additional studies were performed on surrogate hosts Amaranthus caudatus, Ipomoea tricolor, and I. purpurea. Plants of A. caudatus inoculated with B. amyloliquefaciens demonstrated intracellular colonization of guard cells and other epidermal cells, confirming the pattern observed in the orchids. Isolations and histological studies suggest that the bacterium may penetrate deeply into developing plant tissues in shoot meristems, forming endospores in maturing tissues. B. amyloliquefaciens produced fungal inhibitors in culture. In controlled experiments using morning glory seedlings we showed that the bacterium promoted seedling growth and reduced seedling necrosis due to pathogens. We detected the gene for phosphopantetheinyl transferase (sfp), an enzyme in the pathway for production of antifungal lipopeptides, and purified the lipopeptide "surfactin" from cultures of the bacterium. We hypothesize that B. amyloliquefaciens is a robust endophyte and defensive mutualist of vanilla orchids. Whether the symbiosis between this bacterium and its hosts can be managed to protect vanilla crops from diseases is a question that should be evaluated in future research. PMID:25060609

  20. A novel serine metallokeratinase from a newly isolated Bacillus pumilus A1 grown on chicken feather meal: biochemical and molecular characterization.

    PubMed

    Fakhfakh-Zouari, Nahed; Hmidet, Noomen; Haddar, Anissa; Kanoun, Safia; Nasri, Moncef

    2010-09-01

    A keratinolytic enzyme (KerA1) secreted by a newly isolated Bacillus pumilus strain A1 cultivated in medium containing chicken feather meal was purified and characterized, and the gene was isolated and sequenced. The molecular mass of the purified enzyme was estimated to be 34,000 Da by sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis and gel filtration. The optimum pH and temperature for the purified keratinase were 9.0 and 60 degrees C, respectively, using keratin as a substrate. KerA1 showed a high stability towards nonionic surfactants. It was found to be relatively stable toward the strong anionic surfactant (SDS). The deduced amino acid sequence of the keratinase KerA1 differs from both the organic solvent tolerant protease of B. pumilus 115b and the dehairing protease of B. pumilus UN-31-C-42 by one and nine amino acids, respectively. These results suggest that this keratinase may be a useful alternative and ecofriendly route for handling the abundant amount of waste feathers and for applications in detergent formulations. PMID:20012915

  1. Production of a cellulase-free alkaline xylanase from Bacillus pumilus MTCC 5015 by submerged fermentation and its application in biobleaching.

    PubMed

    Thomas, Leya; Sindhu, Raveendran; Binod, Parameswaran; Pandey, Ashok

    2015-06-01

    Here, we described the production of a cellulase-free alkaline xylanase from Bacillus pumilus MTCC 5015 by submerged fermentation and its application in biobleaching. Various process parameters affecting xylanase production by B. pumilus were optimized by adopting a Plackett-Burman design (PBD) as well as Response surface methodology (RSM). These statistical methods aid in improving the enzyme yield by analysing the individual crucial components of the medium. Maximum production was obtained with 4% yeast extract, 0.08% magnesium sulphate, 30 h of inoculum age, incubation temperature of 33.5 degrees C and pH 9.0. Under optimized conditions, the xylanase activity was 372 IU/ml. Media engineering improved a 5-fold increase in the enzyme production. Scanning electron microscopy (SEM) showed significant changes on the surface of xylanase treated pulps as a result of xylan hydrolysis. Increased roughness of paper carton fibres was apparent in scanning electron micrograph due to opening of the micro fibrils present on the surface by xylanase action. The untreated pulp did not show any such change. These results demonstrated that the B. pumilus MTCC 5015 xylanase was effective in bio-bleaching of paper carton. PMID:26155675

  2. Characterization of thermostable serine alkaline protease from an alkaliphilic strain Bacillus pumilus MCAS8 and its applications.

    PubMed

    Jayakumar, Renganathan; Jayashree, Shanmugam; Annapurna, Balumuri; Seshadri, Sundaram

    2012-12-01

    This study describes the characterization and optimization of medium components for an extracellular detergent, surfactant, organic solvent and thermostable serine alkaline protease produced by alkaliphilic Bacillus pumilus MCAS8 strain isolated from Pulicat lake sediments, Tamil Nadu, India. The strain yielded maximum protease (2,214 U/ml) under optimized conditions: carbon source, citric acid-1.5 % (w/w); inducer, soyabean meal-2 % (w/w); pH 11.0; shaking condition 37 °C for 48 h. The enzyme had pH and temperature optima of 9.0 and 60 °C, respectively. The enzyme displayed the molecular mass of 36 kDa in sodium dodecyl sulphate-polyacrylamide gel electrophoresis study and exhibited activity at a wide range of pH (6.0-11.0) and thermostability (20-70 °C). More than 70 % residual activity was observed when the enzyme was incubated with dithiothreitol, ethylenediaminetetraacetic acid, ethylene glycol tetraacetic acid and H(2)O(2) for 30 min. The protease activity was also enhanced by divalent cations such as Ba(2+), Ca(2+) and Mg(2+) and was strongly inhibited by Fe(2+), Zn(2+), Sr(2+), Hg(2+) and urea. The enzyme retained more than 50 % of its initial activity after pre-incubation for 1 h in the presence of 5 % (v/v) organic solvents such as dimethyl sulphoxide and acetone. The protease could hydrolyse various native proteinaceous substrates (1 % w/v) such as bovine serum albumin, casein, skim milk, gelatine, azocasein and haemoglobin. Wash performance analysis of enzyme revealed that it could effectively remove blood stains from the cotton fabric, thus making it suitable to use as an effective detergent additive. The protease enzyme also exhibited promising result in the dehairing of goat skin. The potency of the eco-friendly enzyme without using any chemicals against washing and dehairing showed that the enzyme could be used for various industrial applications. PMID:23076563

  3. Effects of Actin-Like Proteins Encoded by Two Bacillus pumilus Phages on Unstable Lysogeny, Revealed by Genomic Analysis

    PubMed Central

    Yuan, Yihui; Peng, Qin; Wu, Dandan; Kou, Zheng; Wu, Yan; Liu, Pengming

    2014-01-01

    We characterized two newly isolated myoviruses, Bp8p-C and Bp8p-T, infecting the ginger rhizome rot disease pathogen Bacillus pumilus GR8. The plaque of Bp8p-T exhibited a clear center with a turbid rim, suggesting that Bp8p-T could transform into latent phage. Lysogeny assays showed that both the two phages could form latent states, while Bp8p-T could form latent phage at a higher frequency and stability than Bp8p-C. The genomes of Bp8p-C and Bp8p-T were 151,417 and 151,419 bp, respectively; both encoded 212 putative proteins, and only differed by three nucleotides. Moreover, owing to this difference, Bp8p-C encoded a truncated, putative actin-like plasmid segregation protein Gp27-C. Functional analysis of protein Gp27 showed that Gp27-T encoded by Bp8p-T exhibited higher ATPase activity and assembly ability than Gp27-C. The results indicate that the difference in Gp27 affected the phage lysogenic ability. Structural proteome analysis of Bp8p-C virion resulted in the identification of 14 structural proteins, among which a pectin lyase-like protein, a putative poly-gamma-glutamate hydrolase, and three proteins with unknown function, were firstly identified as components of the phage virion. Both phages exhibited specific lytic ability to the host strain GR8. Bp8p-C showed better control effect on the pathogen in ginger rhizome slices than Bp8p-T, suggesting that Bp8p-C has a potential application in bio-control of ginger rhizome rot disease. PMID:25344242

  4. Cloning of the Bacillus pumilus beta-xylosidase gene (xynB) and its expression in Saccharomyces cerevisiae.

    PubMed

    La Grange, D C; Pretorius, I S; van Zyl, W H

    1997-03-01

    A genomic DNA library of the bacterium Bacillus pumilus PLS was constructed and the beta-xylosidase gene (xynB) was amplified from a 3-kb genomic DNA fragment with the aid of the polymerase chain reaction technique. The amplified xynB gene was inserted between the yeast alcohol dehydrogenase II gene promoter (ADH2P) and terminator (ADH2T) sequences on a multicopy episomal plasmid (pDLG11). The xynB gene was also fused in-frame to the secretion signal sequence of the yeast mating pheromone alpha-factor (MF alpha 1S) before insertion between the ADH2P and ADH2T sequences on a similar multicopy episomal plasmid (pDLG12). The resulting construct ADH2P-MF alpha 1S-xynB-ADH2T was designated XLO1. Both plasmids pDLG11 and PDLG12 were introduced into Saccharomyces cerevisiae but only the expression of the XLO1 gene yielded biologically functional beta-xylosidase. The total beta-xylosidase activity remained cell-associated with a maximum activity of 0.09 nkat/ml obtained when the recombinant S. cerevisiae strain was grown for 143 h in synthetic medium. The temperature and pH optima of the recombinant Xlo1 enzyme were 45-50 degrees C and pH 6.6 respectively. The enzyme was thermostable at 45 degrees C; however, at 60 degrees C most of the Xlo1 was inactive after 5 min. PMID:9114518

  5. Immobilization of xylanase from Bacillus pumilus strain MK001 and its application in production of xylo-oligosaccharides.

    PubMed

    Kapoor, Mukesh; Kuhad, Ramesh Chander

    2007-08-01

    Xylanase from Bacillus pumilus strain MK001 was immobilized on different matrices following varied immobilization methods. Entrapment using gelatin (GE) (40.0%), physical adsorption on chitin (CH) (35.0%), ionic binding with Q-sepharose (Q-S) (45.0%), and covalent binding with HP-20 beads (42.0%) showed the maximum xylanase immobilization efficiency. The optimum pH of immobilized xylanase shifted up to 1.0 unit (pH 7.0) as compared to free enzyme (pH 6.0). The immobilized xylanase exhibited higher pH stability (up to 28.0%) in the alkaline pH range (7.0-10.0) as compared to free enzyme. Optimum temperature of immobilized xylanase was observed to be 8 degrees C higher (68.0 degrees C) than free enzyme (60.0 degrees C). The free xylanase retained 50.0% activity, whereas xylanase immobilized on HP-20, Q-S, CH, and GE retained 68.0, 64.0, 58.0, and 57.0% residual activity, respectively, after 3 h of incubation at 80.0 degrees C. The immobilized xylanase registered marginal increase and decrease in Km and Vmax values, respectively, as compared to free enzyme. The immobilized xylanase retained up to 70.0% of its initial hydrolysis activity after seven enzyme reaction cycles. The immobilized xylanase was found to produce higher levels of high-quality xylo-oligosaccharides from birchwood xylan, indicating its potential in the nutraceutical industry. PMID:18025574

  6. Purification and mode of action of an alkali-resistant endo-1, 4-beta-glucanase from Bacillus pumilus.

    PubMed

    Christakopoulos, P; Hatzinikolaou, D G; Fountoukidis, G; Kekos, D; Claeyssens, M; Macris, B J

    1999-04-01

    Alkaline endo-1,4-beta-d-glucanase was secreted by Bacillus pumilus grown in submerged culture on a combination of oat spelt xylan and corn starch as carbon sources. The enzyme was purified to homogeneity by Sephacryl S-200 and Q-Sepharose column chromatography. The protein corresponded to molecular mass and pI values of 67 kDa and 3.7, respectively. The enzyme was optimally active at pH 7.0-8.0 and 60 degrees C and retained 50% of its optimum activity at pH 12. The most notable characteristic of the endoglucanase was its high stability up to pH 12 for 20 h at 30 degrees C. The enzyme hydrolyzed carboxymethylcellulose (CMC) and cello-oligosaccharides but was inactive on cellobiose, cellotriose, Avicel, xylan, 4-nitrophenyl-beta-d-glucoside, 4-nitrophenyl-beta-d-cellobioside, and 4-nitrophenyl-beta-d-xyloside. Analysis of reaction mixtures by HPLC revealed that the enzyme produced almost exclusively cellotriose when acted on CMC and appeared to hydrolyze cello-oligosaccharides by successively releasing cellotriose. The use of 4-methylumbelliferyl cello-oligosaccharides and the determination of bond cleavage frequency revealed that the enzyme preferentially hydrolyzed the third glycosidic bond adjacent to the glycon. The enzyme mediated a decrease in the viscosity of CMC associated with a release of only small amounts of reducing sugar. The enzyme activity was not inhibited by metal ions, surfactants, and chelating agents used as components of laundry detergents. PMID:10087165

  7. Single amino acid mutation alters thermostability of the alkaline protease from Bacillus pumilus: thermodynamics and temperature dependence.

    PubMed

    Huang, Rong; Yang, Qingjun; Feng, Hong

    2015-02-01

    Dehairing alkaline protease (DHAP) from Bacillus pumilus BA06 has been demonstrated to have high catalytic efficiency and good thermostability, with potential application in leather processing. In order to get insights into its catalytic mechanism, two mutants with single amino acid substitution according to the homology modeling and multiple sequence alignment were characterized in thermodynamics of thermal denaturation and temperature dependence of substrate hydrolysis. The results showed that both mutants of V149I and R249E have a systematic increase in catalytic efficiency (kcat/Km) in a wide range of temperatures, mainly due to an increase of k1 (substrate diffusion) and k2 (acylation) for V149I and of k2 and k3 (deacylation) for R249E. In comparison with the wild-type DHAP, the thermostability is increased for V149I and decreased for R249E. Thermodynamic analysis indicated that the free energy (?Ga°) of activation for thermal denaturation may govern the thermostability. The value of ?Ga° is increased for V149I and decreased for R249E. Based on these data and the structural modeling, it is suggested that substitution of Val149 with Ile may disturb the local flexibility in the substrate-binding pocket, leading to enhancement of binding affinity for the substrate. In contrast, substitution of Arg249 with Glu leads to interruption of interaction with the C-terminal of enzyme, thus resulting in less thermostability. This study indicates that amino acid residues in the active center or in the substrate-binding pocket may disturb the catalytic process and can be selected as the target for protein engineering in the bacterial alkaline proteases. PMID:25534779

  8. Effects of actin-like proteins encoded by two Bacillus pumilus phages on unstable lysogeny, revealed by genomic analysis.

    PubMed

    Yuan, Yihui; Peng, Qin; Wu, Dandan; Kou, Zheng; Wu, Yan; Liu, Pengming; Gao, Meiying

    2015-01-01

    We characterized two newly isolated myoviruses, Bp8p-C and Bp8p-T, infecting the ginger rhizome rot disease pathogen Bacillus pumilus GR8. The plaque of Bp8p-T exhibited a clear center with a turbid rim, suggesting that Bp8p-T could transform into latent phage. Lysogeny assays showed that both the two phages could form latent states, while Bp8p-T could form latent phage at a higher frequency and stability than Bp8p-C. The genomes of Bp8p-C and Bp8p-T were 151,417 and 151,419 bp, respectively; both encoded 212 putative proteins, and only differed by three nucleotides. Moreover, owing to this difference, Bp8p-C encoded a truncated, putative actin-like plasmid segregation protein Gp27-C. Functional analysis of protein Gp27 showed that Gp27-T encoded by Bp8p-T exhibited higher ATPase activity and assembly ability than Gp27-C. The results indicate that the difference in Gp27 affected the phage lysogenic ability. Structural proteome analysis of Bp8p-C virion resulted in the identification of 14 structural proteins, among which a pectin lyase-like protein, a putative poly-gamma-glutamate hydrolase, and three proteins with unknown function, were firstly identified as components of the phage virion. Both phages exhibited specific lytic ability to the host strain GR8. Bp8p-C showed better control effect on the pathogen in ginger rhizome slices than Bp8p-T, suggesting that Bp8p-C has a potential application in bio-control of ginger rhizome rot disease. PMID:25344242

  9. Whole-Genome Shotgun Sequence of Bacillus mojavensis Strain RRC101, an Endophytic Bacterium Antagonistic to the Mycotoxigenic Endophytic Fungus Fusrium verticillioides

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Here we report the whole genome shotgun sequence of Bacillus mojavensis strain RRC101, isolated from a maize kernel. This strain is antagonistic to the mycotoxigenic plant pathogen Fusarium verticillioides, and grows within maize tissue, suggesting potential as an endophytic biocontrol agent....

  10. Whole-Genome Shotgun Sequence of Bacillus mojavensis Strain RRC101, an Endophytic Bacterium Antagonistic to the Mycotoxigenic Endophytic Fungus Fusarium verticillioides

    PubMed Central

    Blacutt, A. A.; Meinersmann, R. J.; Bacon, C. W.

    2014-01-01

    Here, we report the whole-genome shotgun sequence of Bacillus mojavensis strain RRC101, isolated from a maize kernel. This strain is antagonistic to the mycotoxigenic plant pathogen Fusarium verticillioides and grows within maize tissue, suggesting potential as an endophytic biocontrol agent. PMID:25359909

  11. The Discovery of phiAGATE, A Novel Phage Infecting Bacillus pumilus, Leads to New Insights into the Phylogeny of the Subfamily Spounavirinae

    PubMed Central

    Barylski, Jakub; Nowicki, Grzegorz; Goździcka-Józefiak, Anna

    2014-01-01

    The Bacillus phage phiAGATE is a novel myovirus isolated from the waters of Lake Góreckie (a eutrophic lake in western Poland). The bacteriophage infects Bacillus pumilus, a bacterium commonly observed in the mentioned reservoir. Analysis of the phiAGATE genome (149844 base pairs) resulted in 204 predicted protein-coding sequences (CDSs), of which 53 could be functionally annotated. Further investigation revealed that the bacteriophage is a member of a previously undescribed cluster of phages (for the purposes of this study we refer to it as “Bastille group”) within the Spounavirinae subfamily. Here we demonstrate that these viruses constitute a distinct branch of the Spounavirinae phylogenetic tree, with limited similarity to phages from the Twortlikevirus and Spounalikevirus genera. The classification of phages from the Bastille group into any currently accepted genus proved extremely difficult, prompting concerns about the validity of the present taxonomic arrangement of the subfamily. PMID:24466180

  12. The discovery of phiAGATE, a novel phage infecting Bacillus pumilus, leads to new insights into the phylogeny of the subfamily Spounavirinae.

    PubMed

    Barylski, Jakub; Nowicki, Grzegorz; Go?dzicka-Józefiak, Anna

    2014-01-01

    The Bacillus phage phiAGATE is a novel myovirus isolated from the waters of Lake Góreckie (a eutrophic lake in western Poland). The bacteriophage infects Bacillus pumilus, a bacterium commonly observed in the mentioned reservoir. Analysis of the phiAGATE genome (149844 base pairs) resulted in 204 predicted protein-coding sequences (CDSs), of which 53 could be functionally annotated. Further investigation revealed that the bacteriophage is a member of a previously undescribed cluster of phages (for the purposes of this study we refer to it as "Bastille group") within the Spounavirinae subfamily. Here we demonstrate that these viruses constitute a distinct branch of the Spounavirinae phylogenetic tree, with limited similarity to phages from the Twortlikevirus and Spounalikevirus genera. The classification of phages from the Bastille group into any currently accepted genus proved extremely difficult, prompting concerns about the validity of the present taxonomic arrangement of the subfamily. PMID:24466180

  13. [Effect of Bacillus pumilus ribonuclease on the paramagnetic centers of microbial cells].

    PubMed

    Zelenikhin, P V; Makeeva, A V; Lozhkin, A P; Rodionov, A A; Nguen, N; Il'inskaia, O N

    2014-01-01

    Potential clinical application of Bacillus pumulus cytotoxic ribonuclease (binase) selectively inducing the death of tumor cells makes it imperative to investigate its effect on the normal human microflora. Flow cytometry was used to determine that binase concentration causing the apoptosis of cancer cells had no effect of the viability of Escherichia coli K12. The changes in the paramagnetic centers of E. coli K12 cells in the presence of nontoxic binase concentrations revealed by EPR spectroscopy included higher EPR signals from iron-containing proteins (including those from the Fe-S clusters) and of the Mn(II) hyperfine structure. The TMTH spin probe (N-(1-hydroxy-2,2,6,6-tetramethylpiperidine-4-il)-2-methylpropanamide hydrochloride) was used to reveal a twofold increase in the levels of reactive oxygen species (ROS) in the cells, which induced oxidative stress in the enzyme-treated bacteria. Inductively coupled plasma mass spectrometry revealed elevated contents of alkaline (Li, Na, K), alkali earth (Mg, Ca), transition (Cr, Mn, Fe, Cu, Zn), and post-transition metals (Bi, Pb) in the cells. Elevated levels of Cu and Zn (which impair the activity of the respiratory chain enzymes) and of Mn, which is known as a superoxide dismutase cofactor, confirmed development of the oxidative stress in bacteria. PMID:25423735

  14. Antitumor activity of bacterial exopolysaccharides from the endophyte Bacillus amyloliquefaciens sp. isolated from Ophiopogon japonicus

    PubMed Central

    CHEN, YI-TAO; YUAN, QIANG; SHAN, LE-TIAN; LIN, MEI-AI; CHENG, DONG-QING; LI, CHANG-YU

    2013-01-01

    The endophytic bacterium, MD-b1, was isolated from the medicinal plant Ophiopogon japonicas and identified as the Bacillus amyloliquefaciens sp. with 99% similarity based on the partial sequence analysis of 16S rDNA. Exopolysaccharides were extracted from the endophyte for the evaluation of its antitumor activity against gastric carcinoma cell lines (MC-4 and SGC-7901). 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays and microscopy were performed to estimate the cell viability and morphological changes of the MC-4 and SGC-7901 cells following treatment with the exopolysaccharides at 14, 22 and 30 ?g/?l. The results revealed that the exopolysaccharides displayed concentration-dependent inhibitory effects against the MC-4 and SGC-7901 cells, with an IC50 of 19.7 and 26.8 ?g/?l, respectively. The exopolysaccharides also induced morphological abnormalities in the cells. These effects indicated the the exopolysaccharides had an antitumoral mechanism of action associated with the mitochondrial dysfunction of the treated cells. This is the first study to investigate the endophytic microorganism isolated from O. japonicas and also the first discovery of such antitumoral exopolysaccharides derived from the genus Bacillus. This provides a promising and reproducible natural product source with high therapeutic value for anticancer treatment, thereby facilitating the development of new anticancer agents. PMID:23833642

  15. Biogenesis of antibacterial silver nanoparticles using the endophytic bacterium Bacillus cereus isolated from Garcinia xanthochymus

    PubMed Central

    Sunkar, Swetha; Nachiyar, C Valli

    2012-01-01

    Objective To synthesize the ecofriendly nanoparticles, which is viewed as an alternative to the chemical method which initiated the use of microbes like bacteria and fungi in their synthesis. Methods The current study uses the endophytic bacterium Bacillus cereus isolated from the Garcinia xanthochymus to synthesize the silver nanoparticles (AgNPs). The AgNPs were synthesized by reduction of silver nitrate solution by the endophytic bacterium after incubation for 3-5 d at room temperature. The synthesis was initially observed by colour change from pale white to brown which was confirmed by UV-Vis spectroscopy. The AgNPs were further characterized using FTIR, SEM-EDX and TEM analyses. Results The synthesized nanoparticles were found to be spherical with the size in the range of 20-40 nm which showed a slight aggregation. The energy-dispersive spectra of the nanoparticle dispersion confirmed the presence of elemental silver. The AgNPs were found to have antibacterial activity against a few pathogenic bacteria like Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, Salmonella typhi and Klebsiella pneumoniae. Conclusions The endophytic bacteria identified as Bacillus cereus was able to synthesize silver nanoparticles with potential antibacterial activity. PMID:23593575

  16. Characterization of boron resistant and accumulating bacteria Lysinibacillus fusiformis M1, Bacillus cereus M2, Bacillus cereus M3, Bacillus pumilus M4 isolated from former mining site, Hokkaido, Japan.

    PubMed

    Raja, Chellaiah Edward; Omine, Kiyoshi

    2012-01-01

    Boron is known to be widespread environmental contaminant that is relatively mobile in soil when compared to other metal contaminants. The present study made an attempt to isolate and characterize the boron resistant and accumulating bacteria from former mining site at Hokkaido, Japan. Four potential strains M1, M2, M3 and M4 were selected based on high degree of boron and heavy metal resistances. The morphological, biochemical and 16S rDNA sequencing analysis of mining bacteria revealed that the isolates were highly homology to Lysinibacillus fusiformis M1 (99 %), Bacillus cereus M2 (99 %), Bacillus cereus M3 (99 %) and Bacillus pumilus M4 (99 %) respectively. The strains M1, M2, M3 and M4 showed resistance to several heavy metals such as As (III), As (V) and Cr (VI), Cu, Ni, Pb and Zn. The selected strains were found to be arsenic oxidizing bacteria confirmed by Silver nitrate test. The resting and growing cells of mining bacteria were used for boron accumulation analysis. Selected strains were found to be efficiently accumulating boron concentration ranging from 0.1-2.3 mg L (-1) and 1.5-4.7 mg L (-1) at 24 h and 168 h, respectively. The following results conclude that the mining bacteria act as potent bioaccumulator of boron and its resistant, removal characteristic can be valuable in boron bioremediation. PMID:22571522

  17. Sensitive change of iso-branched fatty acid (iso-15:0) in Bacillus pumilus PAMC 23174 in response to environmental changes.

    PubMed

    Yi, Da-Hye; Sathiyanarayanan, Ganesan; Seo, Hyung Min; Kim, Jung-Ho; Bhatia, Shashi Kant; Kim, Yun-Gon; Park, Sung-Hee; Jung, Ji-Young; Lee, Yoo Kyung; Yang, Yung-Hun

    2016-01-01

    In this study, the environmental adaptive metabolic processes were investigated using a psychrotrophic polar bacterium Bacillus pumilus PAMC 23174 in response to various temperatures and nutrients, especially in regard to the synthesis of fatty acids. Fatty acid methyl ester analysis was performed using gas chromatography-mass spectrometry and we found that a sensitive changes in iso-branched fatty acid (iso-15:0) synthesis occurred when adjusting the nutritional ratio of branched chain fatty acids (anteiso/iso) with different temperatures, resulting in a change in the balance of anteiso- and iso-form fatty acids. We also observed that this Arctic bacterium preferred amino acid leucine for the synthesis of fatty acids. The increased and decreased synthesis of iso-form fatty acids in response to different temperatures and leucine preference, changes the fatty acid ratio in bacteria, which further affects the membrane fluidity and it is also directly correlated with survival of bacteria in an extreme environment. Hence, this study suggests that B. pumilus PAMC 23174 is a potential model organism for the analysis of the unique ecological adaptations of polar bacteria in changing and the extreme environments. PMID:26566954

  18. Concomitant production, partial purification and characterization of a serine protease and a proteolysis-resistant metallolipase from Bacillus pumilus SG2.

    PubMed

    Sangeetha, Ragupathy; Geetha, Arumugam; Arulpandi, Irulandi

    2010-01-01

    Our objective was to investigate the concomitant production of protease and lipase by a bacterial strain. A promising bacterial strain was isolated from a food-processing industrial effluent, which can produce both protease and lipase. The isolate was characterized by sequencing the 16S rRNA gene. The PCR amplified gene was subjected to analysis by BLAST to ascertain the genetic relatedness of the isolate, Bacillus pumilus SG2. The enzymes were produced and subjected to purification by ammonium sulfate precipitation and dialysis followed by gel filtration chromatography; twelve-fold purity was obtained. The lipase produced was found to be proteolysis-resistant. The partially purified enzymes were characterized for their optimum pH value, temperature, response to inhibitors, surfactants and oxidants. The relative molecular weights of protease and lipase were determined as 28 kDa and 40 kDa, respectively, by zymogram studies. PMID:20355323

  19. Draft Genome Sequence of Bacillus amyloliquefaciens XK-4-1, a Plant Growth-Promoting Endophyte with Antifungal Activity.

    PubMed

    Sun, Zhengxiang; Hsiang, Tom; Zhou, Yi; Zhou, Jinglong

    2015-01-01

    Here, we report the draft genome sequence of a bacterial plant-growth-promoting endophyte, Bacillus amyloliquefaciens XK-4-1, which consists of one circular chromosome of 3,941,805 bp with 3,702 coding sequences (CDSs). The data presented highlight multiple sets of functional genes associated with its plant-beneficial characteristics. PMID:26564038

  20. Characterization of Bacillus subtilis HC8, a novel plant?beneficial endophytic strain from giant hogweed

    PubMed Central

    Malfanova, Natalia; Kamilova, Faina; Validov, Shamil; Shcherbakov, Andrey; Chebotar, Vladimir; Tikhonovich, Igor; Lugtenberg, Ben

    2011-01-01

    Summary Thirty endophytic bacteria were isolated from various plant species growing near Saint?Petersburg, Russia. Based on a screening for various traits, including plant?beneficial properties and DNA fragment patterns, potential siblings were removed. The remaining isolates were taxonomically identified using 16S rDNA sequences and potential human and plant pathogens were removed. The remaining strains were tested for their ability to promote radish root growth and to protect tomato plants against tomato foot and root rot. One strain, Bacillus subtilis HC8, isolated from the giant hogweed Heracleum sosnowskyi Manden, significantly promoted plant growth and protected tomato against tomato foot and root rot. Metabolites possibly responsible for these plant?beneficial properties were identified as the hormone gibberellin and (lipo)peptide antibiotics respectively. The antibiotic properties of strain HC8 are similar to those of the commercially available plant?beneficial strain Bacillus amyloliquefaciens FZB42. However, thin layer chromatography profiles of the two strains differ. It is speculated that endophytes such as B. subtilis HC8 contribute to the fast growth of giant hogweed. PMID:21366893

  1. Effects of direct-fed Bacillus pumilus 8G-134 on feed intake, milk yield, milk composition, feed conversion, and health condition of pre- and postpartum Holstein cows.

    PubMed

    Luan, S; Duersteler, M; Galbraith, E A; Cardoso, F C

    2015-09-01

    The usage of direct-fed microbials (DFM) has become common in the dairy industry, but questions regarding choice of strain, mode of action, and efficacy remain prevalent. The objective of this study was to evaluate the effects of a DFM (Bacillus pumilus 8G-134) on pre- and postpartum performance and incidence of subclinical ketosis in early lactation. Forty-three multiparous Holstein cows were assigned to 2 treatments in a randomized complete block design; cows in the direct-fed microbial treatment (DFMt, n=21) received 5.0×10(9) cfu/cow of B. pumilus in 28 g of a maltodextrin carrier, whereas cows in the control treatment (CON, n=22) received 28 g of maltodextrin carrier alone. Treatments were top-dressed on the total mixed ration daily. Treatments were applied from 21 d before expected calving date to 154 d after calving. Cows on treatment DFMt tended to have lower serum haptoglobin concentration than CON cows on d 14. Cows on treatment DFMt had higher IgA concentrations in milk than CON cows during the first week after calving. Cows fed DFMt had higher yields of milk, fat-corrected milk, energy-corrected milk, milk fat, and milk protein during the second week of lactation than CON; however, we found no differences between treatments on milk yield and milk components overall. Cows on DFMt tended to have higher feed conversion and to have lower prevalence of subclinical ketosis (beta-hydroxybutyrate >1.2 mmol/L) on d 5 than cows fed CON. Dry matter intake, body weight, and body condition score were not affected by DFMt supplementation. Milk production efficiencies (calculated based on fat-corrected milk and energy-corrected milk) were higher by 0.1 kg of milk per kilogram of dry matter intake in cows that received DFMt compared with cows that received CON. In conclusion, cows receiving DFMt tended to have lower incidence of subclinical ketosis than cows receiving CON. Cows fed DFMt tended to have higher feed conversion and evidence for greater immunity than CON. Supplementation with B pumilus 8G-134 may provide benefits for transition cow health and milk production efficiency. PMID:26162791

  2. Isolation, Identification and Characteristics of an Endophytic Quinclorac Degrading Bacterium Bacillus megaterium Q3

    PubMed Central

    Wang, Yunsheng; Zeng, Aiping; Zhou, Xiaomao; Luo, Feng; Bai, Lianyang

    2014-01-01

    In this study, we isolated an endophytic quinclorac-degrading bacterium strain Q3 from the root of tobacco grown in quinclorac contaminated soil. Based on morphological characteristics, Biolog identification, and 16S rDNA sequence analysis, we identified strain Q3 as Bacillus megaterium. We investigated the effects of temperature, pH, inoculation size, and initial quinclorac concentration on growth and degrading efficiency of Q3. Under the optimal degrading condition, Q3 could degrade 93% of quinclorac from the initial concentration of 20 mg/L in seven days. We analyzed the degradation products of quinclorac using liquid chromatography–tandem mass spectrometry (LC-MS/MS). The major degradation products by Q3 were different from those of previously identified quinclorac degrading strains, which suggests that Q3 may employ new pathways for quinclorac degradation. Our indoor pot experiments demonstrated that Q3 can effectively alleviate the quinclorac phytotoxicity in tobacco. As the first endophytic microbial that is capable of degrading quinclorac, Q3 can be a good bioremediation bacterium for quinclorac phytotoxicity. PMID:25244184

  3. Antioxidant and DNA Damage Protecting Activity of Exopolysaccharides from the Endophytic Bacterium Bacillus cereus SZ1.

    PubMed

    Zheng, Li Ping; Zou, Tin; Ma, Yan Jun; Wang, Jian Wen; Zhang, Yu Qing

    2016-01-01

    An endophytic bacterium was isolated from the Chinese medicinal plant Artemisia annua L. The phylogenetic and physiological characterization indicated that the isolate, strain SZ-1, was Bacillus cereus. The endophyte could produce an exopolysaccharide (EPS) at 46 mg/L. The 1,1-diphenyl-2-picrylhydracyl (DPPH) radical scavenging activity of the EPS reached more than 50% at 3-5 mg/mL. The EPS was also effective in scavenging superoxide radical in a concentration dependent fashion with an EC50 value of 2.6 mg/mL. The corresponding EC50 for scavenging hydroxyl radical was 3.1 mg/mL. Moreover, phenanthroline-copper complex-mediated chemiluminescent emission of DNA damage was both inhibited and delayed by EPS. The EPS at 0.7-1.7 mg/mL also protected supercoiled DNA strands in plasmid pBR322 against scission induced by Fenton-mediated hydroxyl radical. The preincubation of PC12 cells with the EPS prior to H₂O₂ exposure increased the cell survival and glutathione (GSH) level and catalase (CAT) activities, and decreased the level of malondialdehyde (MDA) and lactate dehydrogenase (LDH) activity in a dose-dependent manner, suggesting a pronounced protective effect against H₂O₂-induced cytotoxicity. Our study indicated that the EPS could be useful for preventing oxidative DNA damage and cellular oxidation in pharmaceutical and food industries. PMID:26861269

  4. Isolation, identification and characteristics of an endophytic quinclorac degrading bacterium Bacillus megaterium Q3.

    PubMed

    Liu, Min; Luo, Kun; Wang, Yunsheng; Zeng, Aiping; Zhou, Xiaomao; Luo, Feng; Bai, Lianyang

    2014-01-01

    In this study, we isolated an endophytic quinclorac-degrading bacterium strain Q3 from the root of tobacco grown in quinclorac contaminated soil. Based on morphological characteristics, Biolog identification, and 16S rDNA sequence analysis, we identified strain Q3 as Bacillus megaterium. We investigated the effects of temperature, pH, inoculation size, and initial quinclorac concentration on growth and degrading efficiency of Q3. Under the optimal degrading condition, Q3 could degrade 93% of quinclorac from the initial concentration of 20 mg/L in seven days. We analyzed the degradation products of quinclorac using liquid chromatography-tandem mass spectrometry (LC-MS/MS). The major degradation products by Q3 were different from those of previously identified quinclorac degrading strains, which suggests that Q3 may employ new pathways for quinclorac degradation. Our indoor pot experiments demonstrated that Q3 can effectively alleviate the quinclorac phytotoxicity in tobacco. As the first endophytic microbial that is capable of degrading quinclorac, Q3 can be a good bioremediation bacterium for quinclorac phytotoxicity. PMID:25244184

  5. Development of a high-efficient transformation system of Bacillus pumilus strain DX01 to facilitate gene isolation via gfp-tagged insertional mutagenesis and visualize bacterial colonization of rice roots.

    PubMed

    Shen, Xinqian; Chen, Yunpeng; Liu, Tong; Hu, Xiaolu; Gu, Zhenfang

    2013-09-01

    A Tn5 transposition vector, pMOD-tet-egfp, was constructed and used for the random insertional mutagenesis of Bacillus pumilus. Various parameters were investigated to increase the transformation efficiency B. pumilus DX01 via Tn5 transposition complexes (transposome): bacterial growth phase, type of electroporation buffer, electric field strength, and recovery medium. Transformation efficiency was up to 3 × 10(4)?transformants/?g of DNA under the optimized electroporation conditions, and a total of 1,467 gfp-tagged transformants were obtained. Fluorescence-activated cell sorting analysis showed that all gfp-tagged bacterial cells expressed GFP, indicating that foreign DNA has been successfully integrated into the genome of B. pumilus and expressed. Finally, flanking DNA sequences were isolated from several transformants and colonization of rice roots by B. pumilus DX01 was also studied. The method developed here will be useful for creating an insertion mutant library of gram-positive bacteria, thus facilitating their molecular genetic and cytological studies. PMID:23334949

  6. The overexpression of the SAPB of Bacillus pumilus CBS and mutated sapB-L31I/T33S/N99Y alkaline proteases in Bacillus subtilis DB430: new attractive properties for the mutant enzyme.

    PubMed

    Jaouadi, Nadia Zaraî; Jaouadi, Bassem; Aghajari, Nushin; Bejar, Samir

    2012-02-01

    The sapB gene encoding for Bacillus pumilus CBS protease (SAPB) and the triple mutated sapB-L31I/T33S/N99Y gene were cloned and overexpressed in the protease-deficient Bacillus subtilis DB430 using an Escherichia coli-Bacillus shuttle vector pBSMuL2. The 34,625.13 and 34,675.11-Da enzymes were purified from the culture supernatant of B. subtilis expressing the wild-type and mutated genes, respectively. The purified proteases showed the same N-terminal sequences and biochemical properties of those expressed in E. coli. Further investigations demonstrated that, compared to wild-type and other proteases, SAPB-L31I/T33S/N99Y had the highest catalytic efficiency and the best degree of hydrolysis. The mutant enzyme was also noted to exhibit a number of newly explored properties that are highly valued in the marketplace, namely considerable stability to detergents, higher resistance towards organic solvents, and potent dehairing ability. Overall, the findings indicated that SAPB-L31I/T33S/N99Y is a promising candidate for future use in a wide range of industrial and commercial applications. PMID:22178490

  7. Aqueous extracts of a Mars analogue regolith that mimics the Phoenix landing site do not inhibit spore germination or growth of model spacecraft contaminants Bacillus subtilis 168 and Bacillus pumilus SAFR-032

    NASA Astrophysics Data System (ADS)

    Nicholson, Wayne L.; McCoy, Lashelle E.; Kerney, Krystal R.; Ming, Douglas W.; Golden, D. C.; Schuerger, Andrew C.

    2012-08-01

    Because Mars is a primary target for life detection and habitability assessment missions, its exploration is also by necessity a Planetary Protection issue. The recent finding of significant levels of perchlorate (ClO4-) in regolith sampled from the Phoenix landing site raises the question of its potential biotoxicity to putative indigenous martian life, microbial forward contaminants from Earth, or future human visitors. To address this issue, an analogue regolith was constructed based on regolith chemistry data from the Phoenix landing site. A Mars Aqueous Regolith Extract (MARE) was prepared from the Phoenix analogue regolith and analyzed by ion chromatography. The MARE contained (mg/L) the cations Na+ (1411 ± 181), Mg2+ (1051 ± 160), Ca2+ (832 ± 125), and K+ (261 ± 29), and the anions SO42-(5911±993), ClO4-(5316±1767), Cl(171±25) and F- (2.0 ± 0.4). Nitrogen-containing species NO3-(773±113) and NO2-(6.9±2.3) were also present as a result of regolith preparation procedures, but their relevance to Mars is at present unknown. The MARE was tested for potential toxic effects on two model spacecraft contaminants, the spore-forming bacteria Bacillus subtilis strain 168 and Bacillus pumilus strain SAFR-032. In B. subtilis, spore germination and initial vegetative growth (up to ˜5 h) was not inhibited in a rich complex medium prepared with the MARE, but growth after 5 h was significantly suppressed in medium prepared using the MARE. Both B. subtilis and B. pumilus exhibited significantly higher rates of spore germination and growth in the MARE vs. DW with no additions (likely due to endogenous spore nutrients), but germination and growth was further stimulated by addition of glucose and a combination of buffered inorganic salts (K2HPO4, KH2PO4, (NH4)2SO4, and MgSO4). The data indicate that the aqueous environment in the regolith from the Phoenix landing site containing high levels of perchlorate does not pose a significant barrier to growth of putative forward contaminants such as B. subtilis and B. pumilus under Earth laboratory conditions.

  8. [The role of the promoter structure in the efficiency of the expression of guanylspecific ribonucleases from Bacillus intermedius and Bacillus pumilus].

    PubMed

    Kharitonova, M A; Vershinina, V I; Morozova, O V; Znamenskaia, L V

    2006-01-01

    Guanylspecific ribonucleases from B. intermedius (binase) and B.pumilus (RNase Bpu) are structural and functional homologues, and their biosynthesis is subjected to the same laws. At the same time, there are essential differences in the expression efficiency of binase and RNase Bpu genes. This was first suggested to be due to differences in nucleotide sequences of promoters of the genes. Therefore, we constructed plasmids changing each different nucleotide in binase promoter for corresponding one from RNase Bpu and vise versa. It was found that the difference in RNase Bpu and binase expression was due to the only nucleotide in RNase Bpu promoter. PMID:17094653

  9. Lipase production from a novel thermo-tolerant and extreme acidophile Bacillus pumilus using palm oil as the substrate and treatment of palm oil-containing wastewater.

    PubMed

    Saranya, P; Sukanya Kumari, H; Prasad Rao, B; Sekaran, G

    2014-03-01

    The thermo-tolerant and extreme acidophilic microorganism Bacillus pumilus was isolated from the soil collected from a commercial edible-oil extraction industry. Optimisation of conditions for the lipase production was conducted using response surface methodology. The optimum conditions for obtaining the maximum activity (1,100 U/mL) of extremely acidic thermostable lipase were fermentation time, 96 h; pH, 1; temperature, 50 °C; and concentration of palm oil, 50 g/L. After purification, a 7.1-fold purity of lipase with specific activity of 5,173 U/mg protein was obtained. The molecular weight of the thermo-tolerant acidophilic lipase (TAL) was 55 kDa. The predominant amino acid in the TAL was glycine. The functional groups of lipase were determined by Fourier transform infrared spectroscopy. TAL exhibited enhanced activity (114 %) with dimethyl sulphoxide (20 %, v/v), and it showed a moderate activity with methanol, hexane and benzene. The optimum conditions for the treatment of palm oil in wastewater using the TAL were found to be time, 3 h; pH, 1; temperature, 50 °C with pseudo second-order kinetic constant of 1.88?×?10(-3) L mol(-1) min(-1). The Michaelis-Menten enzyme kinetic model and the nonlinear kinetic model were evaluated for the TAL. TAL established hydrolysis efficiency of 96 % for palm oil in wastewater at 50 °C. PMID:24293300

  10. Purification and characterization of [Formula: see text]-mannanase from Bacillus pumilus (M27) and its applications in some fruit juices.

    PubMed

    Adiguzel, Ahmet; Nadaroglu, Hayrunnisa; Adiguzel, Gulsah

    2015-08-01

    Thermo alkaline mannanase was purified from the bacteria of Bacillus pumilus (M27) using the techniques of ammonium sulphate precipitation, DEAE-Sephadex ion exchange chromatography and Sephacryl S200 gel filtration chromatography with 111-fold and 36 % yield. It was determined that the enzyme had 2 sub-units including 35 kDa and 55 kDa in gel filtration chromatography and SDS-PAGE electrophoresis systems. The optimum pH and temperature was determined as 8 and 60 °C, respectively. It was also noticed that the enzyme did not lose its activity at a wide interval such as pH 3-11 and at high temperatures such as 90 °C. Additionally, the effects of some metal ions on the mannanase enzyme activity. Moreover, the clarifying efficiency of purified mannanase enzyme with some fruit juices such as orange, apricot, grape and apple was also investigated. Enzymatic treatment was carried out with 1 mL L(-1) of purified mannanase for 1 h at 60 °C. It was determined that the highest pure enzyme was efficient upon clarifying the apple juice at 154 % rate. PMID:26243955

  11. Scale-up of an alkaline protease from Bacillus pumilus MTCC 7514 utilizing fish meal as a sole source of nutrients.

    PubMed

    Gupta, Rishikesh Kumar; Prasad, Dinesh; Sathesh, Jaykumar; Naidu, Ramachandra Boopathy; Kamini, Numbi Ramudu; Palanivel, Saravanan; Gowthaman, Marichetti Kuppuswami

    2012-09-01

    Fish meal grades SL1 and SL2 from Sardine (Sardinella longiceps) and NJ from Pink Perch (Nemipterus japonicas) were evaluated as a sole source of carbon and nitrogen in the medium for alkaline protease production by Bacillus pumilus MTCC 7514. The analysis of the fish meal suggests that the carbon and nitrogen contents in fish meal are sufficient to justify its choice as replacement for other nutrients. Protease production increased significantly (4,914 U/ml) in medium containing only fish meal, compared with the basal medium (2,646 U/ml). However, the elimination of inorganic salts from media reduced the protease productivity. In addition, all the three grades of fish meal yielded almost the same amounts of protease when employed as the sole source of carbon and nitrogen. Nevertheless, the best results were observed in fish meal SL1 medium. Furthermore, protease production was enhanced to 6,966 U/ml and 7,047 U/ml on scaling up from flask (4,914 U/ml) to 3.7 and 20 L fermenters, respectively, using fish meal (10 g/l). Similarly, the corresponding improvement in productivities over flask (102.38 U/ml/h) was 193.5 and 195.75 U/ml/h in 3.7 and 20 L fermenters, respectively. The crude protease was found to have dehairing ability in leather processing, which is bound to have great environmental benefits. PMID:22814497

  12. A novel thermostable GH5_7 ?-mannanase from Bacillus pumilus GBSW19 and its application in manno-oligosaccharides (MOS) production.

    PubMed

    Zang, Haoyu; Xie, Shanshan; Wu, Huijun; Wang, Weiduo; Shao, Xiankun; Wu, Liming; Rajer, Faheem Uddin; Gao, Xuewen

    2015-10-01

    A novel thermostable mannanase from a newly isolated Bacillus pumilus GBSW19 has been identified, expressed, purified and characterized. The enzyme shows a structure comprising a 28 amino acid signal peptide, a glycoside hydrolase family 5 (GH5) catalytic domain and no carbohydrate-binding module. The recombinant mannanase has molecular weight of 45 kDa with an optimal pH around 6.5 and is stable in the range from pH 5-11. Meanwhile, the optimal temperature is around 65 °C, and it retains 50% relative activity at 60 °C for 12h. In addition, the purified enzyme can be activated by several ions and organic solvents and is resistant to detergents. Bpman5 can efficiently convert locus bean gum to mainly M2, M3 and M5, and hydrolyze manno-oligosaccharides with a minimum DP of 3. Further exploration of the optimum condition using HPLC to prepare oligosaccharides from locust bean gum was obtained as 10mg/ml locust bean gum incubated with 10 U/mg enzyme at 50 °C for 24h. By using this enzyme, locust bean gum can be utilized to generate high value-added oligosaccharides with a DP of 2-6. PMID:26215338

  13. Sugarcane bagasse as support for immobilization of Bacillus pumilus HZ-2 and its use in bioremediation of mesotrione-contaminated soils.

    PubMed

    Liu, Jie; Chen, Shaohua; Ding, Jie; Xiao, Ying; Han, Haitao; Zhong, Guohua

    2015-12-01

    The degrading microorganisms isolated from environment usually fail to degrade pollutants when used for bioremediation of contaminated soils; thus, additional treatments are needed to enhance biodegradation. In the present study, the potential of sugarcane bagasse as bacteria-immobilizing support was investigated in mesotrione biodegradation. A novel isolate Bacillus pumilus HZ-2 was applied in bacterial immobilization, which was capable of degrading over 95 % of mesotrione at initial concentrations ranging from 25 to 200 mg L(-1) within 4 days in flask-shaking tests. Scanning electron microscope (SEM) images showed that the bacterial cells were strongly absorbed and fully dispersed on bagasse surface after immobilization. Specially, 86.5 and 82.9 % of mesotrione was eliminated by bacteria immobilized on bagasse of 100 and 60 mesh, respectively, which indicated that this immobilization was able to maintain a high degrading activity of the bacteria. Analysis of the degradation products determined 2-amino-4-methylsulfonylbenzoic acid (AMBA) and 4-methylsulfonyl-2-nitrobenzoic acid (MNBA) as the main metabolites in the biodegradation pathway of mesotrione. In the sterile soil, approximately 90 % of mesotrione was degraded after supplementing 5.0 % of molasses in bacteria-bagasse composite, which greatly enhanced microbial adaptability and growth in the soil environment. In the field tests, over 75 % of mesotrione in soil was degraded within 14 days. The immobilized preparation demonstrated that mesotrione could be degraded at a wide range of pH values (5.0-8.0) and temperatures (25-35 °C), especially at low concentrations of mesotrione (5 to 20 mg kg(-1)). These results showed that sugarcane bagasse might be a good candidate as bacteria-immobilizing support to enhance mesotrione degradation by Bacillus p. HZ-2 in contaminated soils. PMID:26337896

  14. Catalytic versatility of Bacillus pumilus beta-xylosidase: glycosyl transfer and hydrolysis promoted with alpha- and beta-D-xylosyl fluoride.

    PubMed

    Kasumi, T; Tsumuraya, Y; Brewer, C F; Kersters-Hilderson, H; Claeyssens, M; Hehre, E J

    1987-06-01

    Bacillus pumilus beta-xylosidase, an enzyme considered restricted to hydrolyzing a narrow range of beta-D-xylosidic substrates with inversion of configuration, was found to catalyze different stereochemical, essentially irreversible, glycosylation reactions with alpha- and beta-D-xylopyranosyl fluoride. The enzyme promoted the hydrolysis of beta-D-xylopyranosyl fluoride at a high rate, V = 6.25 mumol min-1 mg-1 at 0 degrees C, in a reaction that obeyed Michaelis-Menten kinetics. In contrast, its action upon alpha-D-xylopyranosyl fluoride was slow and characterized by an unusual relation between the rate of fluoride release and the substrate concentration, suggesting the possible need for two substrate molecules to be bound at the active center in order for reaction to occur. Moreover, 1H NMR spectra of a digest of alpha-D-xylosyl fluoride showed the substrate to be specifically converted to alpha-D-xylose by the enzyme. The observed retention of configuration is not consistent with direct hydrolysis by this "inverting" enzyme but is strongly indicative of the occurrence of two successive inverting reactions: xylosyl transfer from alpha-D-xylosyl fluoride to form a beta-D-xylosidic product, followed by hydrolysis of the latter to produce alpha-D-xylose. The transient intermediate product formed enzymically from alpha-D-xylosyl fluoride in the presence of [14C]xylose was isolated and shown by its specific radioactivity and 1H NMR spectrum as well as by methylation and enzymic analyses to be 4-O-beta-D-xylopyranosyl-D-xylopyranose containing one [14C]xylose residue.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:3111528

  15. Catalytic versatility of Bacillus pumilus. beta. -xylosidase: glycosyl transfer and hydrolysis promoted with. cap alpha. - and. beta. -D-xylosyl fluoride

    SciTech Connect

    Kasumi, T.; Tsumuraya, Y.; Brewer, C.F.; Kersters-Hilderson, H.; Claeyssens, M.; Hehre, E.J.

    1987-06-02

    Bacillus pumilus ..beta..-xylosidase, an enzyme considered restricted to hydrolyzing a narrow range of ..beta..-D-xylosidic substrates with inversion of configuration, was found to catalyze different stereochemical, essentially irreversible, glycosylation reactions with ..cap alpha..- and ..beta..-D-xylopyranosyl fluoride. The enzyme promoted the hydrolysis of ..beta..-D-xylopyranosyl fluoride at a high rate, V = 6.25 ..mu..mol min/sup -1/ mg/sup -1/ at 0/sup 0/C, in a reaction that obeyed Michaelis-Menten kinetics. In contrast, its action upon ..cap alpha..-D-xylopyranosyl fluoride was slow and characterized by an unusual relation between the rate of fluoride release and the substrate concentration, suggesting the possible need for two substrate molecules to be bound at the active center in order for reaction to occur. Moreover, /sup 1/H NMR spectra of a digest of ..cap alpha..-D-xylosyl fluoride showed the substrate to be specifically converted to ..cap alpha..-D-xylose by the enzyme. The observed retention of configuration is not consistent with direct hydrolysis by this inverting enzyme but is strongly indicative of the occurrence of two successive inverting reactions: xylosyl transfer from ..cap alpha..-D-xylosyl fluoride to form a ..beta..-D-xylosidic product, followed by hydrolysis of the latter to produce ..cap alpha..-D-xylose. The transient intermediate product formed enzymically from ..cap alpha..-D-xylosyl fluoride in the presence of (/sup 14/C)xylose was isolated and shown by its specific radioactivity and /sup 1/H NMR spectrum as well as by methylatino and enzymic analyses to be 4-O-..beta..-D-xylopyranosyl-D-xylopyranose containing one (/sup 14/C)xylose residue.

  16. Isolation and identification of bacterial endophytes from pharmaceutical agarwood-producing Aquilaria species

    PubMed Central

    Bhore, Subhash J.; Preveena, Jagadesan; Kandasamy, Kodi I.

    2013-01-01

    Background: Resins and gums are used in traditional medicine and do have potential applications in pharmacy and medicine. Agarwood is the fragrant resinous wood, which is an important commodity from Aquilaria species and has been used as a sedative, analgesic, and digestive in traditional medicine. Endophytic bacteria are potentially important in producing pharmaceutical compounds found in the plants. Hence, it was important to understand which types of endophytic bacteria are associated with pharmaceutical agarwood-producing Aquilaria species. Objective: This study was undertaken to isolate and identify endophytic bacteria associated with agarwood-producing seven (7) Aquilaria species from Malaysia. Materials and Methods: Botanical samples of seven Aquilaria species were collected, and endophytic bacteria were isolated from surface-sterilized-tissue samples. The 16S rRNA gene fragments were amplified using PCR method, and endophytic bacterial isolates (EBIs) were identified based on 16S rRNA gene sequence similarity based method. Results: Culturable, 77 EBIs were analyzed, and results of 16S rRNA gene sequences analysis suggest that 18 different types of endophytic bacteria are associated with (seven) Aquilaria species. From 77 EBIs, majority (36.4%) of the isolates were of Bacillus pumilus. Conclusion: These findings indicate that agarwood-producing Aquilaria species are harboring 18 different types of culturable endophytic bacteria. PMID:23798890

  17. The combined effect of pasteurization intensity, water activity, pH and incubation temperature on the survival and outgrowth of spores of Bacillus cereus and Bacillus pumilus in artificial media and food products.

    PubMed

    Samapundo, S; Heyndrickx, M; Xhaferi, R; de Baenst, I; Devlieghere, F

    2014-07-01

    The objective of the study was to evaluate the combined effects of pasteurization intensity (no heat treatment and 10 min at 70, 80 and 90 °C), water activity (aw) (0.960-0.990), pH (5.5-7.0) and storage temperature (7 and 10 °C) on the survival and outgrowth of psychrotolerant spores of Bacillus cereus FF119b and Bacillus pumilus FF128a. The experiments were performed in both artificial media and a validation was performed on real food products (cream, béchamel sauce and mixed vegetable soup). It was determined that in general, heat treatments of 10 min at 70 °C or 80 °C activated the spores of both B. cereus FF119b and B. pumilus FF128a, resulting in faster outgrowth compared to native (non-heat treated) spores. A pasteurization treatment of 10 min at 90 °C generally resulted in the longest lag periods before outgrowth of both isolates. Some of the spores were inactivated by this heat treatment, with more inactivation being observed the lower the pH value of the heating medium. Despite this, it was also observed that under some conditions the remaining (surviving) spores were actually activated as their outgrowth took place after a shorter period of time compared to native non-heated spores. While the response of B. cereus FF119b to the pasteurization intensity in cream and béchamel sauce was similar to the trends observed in the artificial media at 10 °C, in difference, outgrowth was only observed at 7 °C in both products when the spores had been heated for 10 min at 80 °C. Moreover, no inactivation was observed in cream or béchamel sauce when the spores were heated for 10 min at 90 °C in these two products. This was attributed to the protective effect of fat in the cream and the ingredients in the béchamel sauce. The study provides some insight into the potential microbial (stability and safety) consequences of the current trend towards milder heat treatments which is being pursued in the food industry. PMID:24801270

  18. Probing the crucial role of Leu31 and Thr33 of the Bacillus pumilus CBS alkaline protease in substrate recognition and enzymatic depilation of animal hide.

    PubMed

    Zaraî Jaouadi, Nadia; Jaouadi, Bassem; Ben Hlima, Hajer; Rekik, Hatem; Belhoul, Mouna; Hmidi, Maher; Ben Aicha, Houda Slimene; Hila, Chiraz Gorgi; Toumi, Abdessatar; Aghajari, Nushin; Bejar, Samir

    2014-01-01

    The sapB gene, encoding Bacillus pumilus CBS protease, and seven mutated genes (sapB-L31I, sapB-T33S, sapB-N99Y, sapB-L31I/T33S, sapB-L31I/N99Y, sapB-T33S/N99Y, and sapB-L31I/T33S/N99Y) were overexpressed in protease-deficient Bacillus subtilis DB430 and purified to homogeneity. SAPB-N99Y and rSAPB displayed the highest levels of keratinolytic activity, hydrolysis efficiency, and enzymatic depilation. Interestingly, and at the semi-industrial scale, rSAPB efficiently removed the hair of goat hides within a short time interval of 8 h, thus offering a promising opportunity for the attainment of a lime and sulphide-free depilation process. The efficacy of the process was supported by submitting depilated pelts and dyed crusts to scanning electron microscopic analysis, and the results showed well opened fibre bundles and no apparent damage to the collagen layer. The findings also revealed better physico-chemical properties and less effluent loads, which further confirmed the potential candidacy of the rSAPB enzyme for application in the leather industry to attain an ecofriendly process of animal hide depilation. More interestingly, the findings on the substrate specificity and kinetic properties of the enzyme using the synthetic peptide para-nitroanilide revealed strong preferences for an aliphatic amino-acid (valine) at position P1 for keratinases and an aromatic amino-acid (phenylalanine) at positions P1/P4 for subtilisins. Molecular modeling suggested the potential involvement of a Leu31 residue in a network of hydrophobic interactions, which could have shaped the S4 substrate binding site. The latter could be enlarged by mutating L31I, fitting more easily in position P4 than a phenylalanine residue. The molecular modeling of SAPB-T33S showed a potential S2 subside widening by a T33S mutation, thus suggesting its importance in substrate specificity. PMID:25264614

  19. Probing the Crucial Role of Leu31 and Thr33 of the Bacillus pumilus CBS Alkaline Protease in Substrate Recognition and Enzymatic Depilation of Animal Hide

    PubMed Central

    Zaraî Jaouadi, Nadia; Jaouadi, Bassem; Ben Hlima, Hajer; Rekik, Hatem; Belhoul, Mouna; Hmidi, Maher; Aicha, Houda Slimene Ben; Hila, Chiraz Gorgi; Toumi, Abdessatar; Aghajari, Nushin; Bejar, Samir

    2014-01-01

    The sapB gene, encoding Bacillus pumilus CBS protease, and seven mutated genes (sapB-L31I, sapB-T33S, sapB-N99Y, sapB-L31I/T33S, sapB-L31I/N99Y, sapB-T33S/N99Y, and sapB-L31I/T33S/N99Y) were overexpressed in protease-deficient Bacillus subtilis DB430 and purified to homogeneity. SAPB-N99Y and rSAPB displayed the highest levels of keratinolytic activity, hydrolysis efficiency, and enzymatic depilation. Interestingly, and at the semi-industrial scale, rSAPB efficiently removed the hair of goat hides within a short time interval of 8 h, thus offering a promising opportunity for the attainment of a lime and sulphide-free depilation process. The efficacy of the process was supported by submitting depilated pelts and dyed crusts to scanning electron microscopic analysis, and the results showed well opened fibre bundles and no apparent damage to the collagen layer. The findings also revealed better physico-chemical properties and less effluent loads, which further confirmed the potential candidacy of the rSAPB enzyme for application in the leather industry to attain an ecofriendly process of animal hide depilation. More interestingly, the findings on the substrate specificity and kinetic properties of the enzyme using the synthetic peptide para-nitroanilide revealed strong preferences for an aliphatic amino-acid (valine) at position P1 for keratinases and an aromatic amino-acid (phenylalanine) at positions P1/P4 for subtilisins. Molecular modeling suggested the potential involvement of a Leu31 residue in a network of hydrophobic interactions, which could have shaped the S4 substrate binding site. The latter could be enlarged by mutating L31I, fitting more easily in position P4 than a phenylalanine residue. The molecular modeling of SAPB-T33S showed a potential S2 subside widening by a T33S mutation, thus suggesting its importance in substrate specificity. PMID:25264614

  20. Isolation and characterization of Leu[7]-Surfactin from the endophytic bacterium Bacillus mojavensis RRC 101, a biocontrol agent for Fusarium verticillioides

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Bacillus mojavensis is an endophytic bacterium patented for control of fungal diseases in maize and other plants. Culture extracts and filtrates from this bacterium were antagonistic to the pathogenic and mycotoxic fungus Fusarium verticillioides. However, the identity of the inhibitory substance ...

  1. Potential for Control of Seedling Blight of Wheat Caused by Fusarium graminearum and Related Species Using the Bacterial Endophyte Bacillus mojavensis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Fusarium infected-wheat seed decreases germination, seedling emergence, and causes post emergence seedling death, and can contribute to wheat scab and ear rot of maize, with consequent production of mycotoxins such as deoxynivalenol and zearalenone. A patented endophytic bacterial strain, Bacillus ...

  2. Formulations of the endophytic bacterium Bacillus subtilis Tu-100 suppress Sclerotinia sclerotiorum on oilseed rape and improve plant vigor in field trials conducted at separate locations

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Sclerotinia sclerotiorum causes serious yield losses in crops in The People’s Republic of China. Two formulations of oilseed rape seed containing the endophytic bacterium Bacillus subtilis Tu-100 were evaluated for suppression of this pathogen in field trials conducted at two independent locations....

  3. The effect of different growth regimes on the endophytic bacterial communities of the fern, Dicksonia sellowiana hook (Dicksoniaceae)

    PubMed Central

    de Araújo Barros, Irene; Luiz Araújo, Welington; Lúcio Azevedo, João

    2010-01-01

    Endophytic bacteria associated with the fern Dicksonia sellowiana were investigated. The bacterial communities from the surface-sterilized pinnae and rachis segments of the plants from the Brazilian Atlantic Rainforest that grew in native field conditions were compared with the bacterial communities from plants grown in greenhouses and plants that were initially grown in greenhouses and then transferred to the forest. From 540 pinnae and 540 rachis segments, 163 (30.2%) and 346 (64.2%) were colonized by bacteria, respectively. The main bacterial genera and species that were isolated included Bacillus spp. ( B. cereus, B. megaterium, B. pumilus and B. subtilis ) , Paenibacillus sp. , Amphibacillus sp. , Gracilibacillus sp. , Micrococcus sp. and Stenotrophomonas spp. ( S. maltophilia and S. nitroreducens ). B. pumilus was the most frequently isolated bacterial species . Amphibacillus and Gracilibacillus were reported as endophytes for the first time. Other commonly found bacterial genera were not observed in D. sellowiana , which may reflect preferences of specific bacterial communities inside this fern or detection limitations due to the isolation procedures. Plants that were grown in greenhouses and plants that were reintroduced into the forest displayed more bacterial genera and species diversity than native field plants, suggesting that reintroduction shifts the bacterial diversity. Endophytic bacteria that displayed antagonistic properties against different microorganisms were detected, but no obvious correlation was found between their frequencies with plant tissues or with plants from different growth regimes. This paper reports the first isolation of endophytic bacteria from a fern. PMID:24031575

  4. The effect of different growth regimes on the endophytic bacterial communities of the fern, Dicksonia sellowiana hook (Dicksoniaceae).

    PubMed

    de Araújo Barros, Irene; Luiz Araújo, Welington; Lúcio Azevedo, João

    2010-10-01

    Endophytic bacteria associated with the fern Dicksonia sellowiana were investigated. The bacterial communities from the surface-sterilized pinnae and rachis segments of the plants from the Brazilian Atlantic Rainforest that grew in native field conditions were compared with the bacterial communities from plants grown in greenhouses and plants that were initially grown in greenhouses and then transferred to the forest. From 540 pinnae and 540 rachis segments, 163 (30.2%) and 346 (64.2%) were colonized by bacteria, respectively. The main bacterial genera and species that were isolated included Bacillus spp. ( B. cereus, B. megaterium, B. pumilus and B. subtilis ) , Paenibacillus sp. , Amphibacillus sp. , Gracilibacillus sp. , Micrococcus sp. and Stenotrophomonas spp. ( S. maltophilia and S. nitroreducens ). B. pumilus was the most frequently isolated bacterial species . Amphibacillus and Gracilibacillus were reported as endophytes for the first time. Other commonly found bacterial genera were not observed in D. sellowiana , which may reflect preferences of specific bacterial communities inside this fern or detection limitations due to the isolation procedures. Plants that were grown in greenhouses and plants that were reintroduced into the forest displayed more bacterial genera and species diversity than native field plants, suggesting that reintroduction shifts the bacterial diversity. Endophytic bacteria that displayed antagonistic properties against different microorganisms were detected, but no obvious correlation was found between their frequencies with plant tissues or with plants from different growth regimes. This paper reports the first isolation of endophytic bacteria from a fern. PMID:24031575

  5. Growth and (137)Cs uptake of four Brassica species influenced by inoculation with a plant growth-promoting rhizobacterium Bacillus pumilus in three contaminated farmlands in Fukushima prefecture, Japan.

    PubMed

    Aung, Han Phyo; Djedidi, Salem; Oo, Aung Zaw; Aye, Yi Swe; Yokoyama, Tadashi; Suzuki, Sohzoh; Sekimoto, Hitoshi; Bellingrath-Kimura, Sonoko Dorothea

    2015-07-15

    The effectiveness of the plant growth-promoting rhizobacterium Bacillus pumilus regarding growth promotion and radiocesium ((137)Cs) uptake was evaluated in four Brassica species grown on different (137)Cs contaminated farmlands at Fukushima prefecture in Japan from June to August 2012. B. pumilus inoculation did not enhance growth in any of the plants, although it resulted in a significant increase of (137)Cs concentration and higher (137)Cs transfer from the soil to plants. The Brassica species exhibited different (137)Cs uptake abilities in the order Komatsuna>turnip>mustard>radish. TF values of (137)Cs ranged from 0.018 to 0.069 for all vegetables. Komatsuna possessed the largest root surface area and root volume, and showed a higher (137)Cs concentration in plant tissue and higher (137)Cs TF values (0.060) than the other vegetables. Higher (137)Cs transfer to plants was prominent in soil with a high amount of organic matter and an Al-vermiculite clay mineral type. PMID:25847170

  6. Diversity of culturable bacterial endophytes of saffron in Kashmir, India.

    PubMed

    Sharma, Tanwi; Kaul, Sanjana; Dhar, Manoj K

    2015-01-01

    Saffron (Crocus sativus) is a medicinally important plant. The Kashmir valley (J&K, India) emblematizes one of the major and quality saffron producing areas in the world. Nonetheless, the area has been experiencing a declining trend in the production of saffron during the last decade. Poor disease management is one of the major reasons for declining saffron production in the area. Endophytes are known to offer control against many diseases of host plant. During the present study, culturable bacterial endophytes were isolated from saffron plant, identified and assessed for plant growth promoting activities. Molecular and phylogenetic analysis grouped the fifty-four bacterial isolates into eleven different taxa, viz. Bacillus licheniformis, B. subtilis, B. cereus, B. humi, B. pumilus, Paenibacillus elgii, B. safensis, Brevibacillus sp., Pseudomonas putida, Staphylococcus hominis and Enterobacter cloacae. The results were also supported with the identification based on BIOLOG system. B. licheniformis was the dominant endophyte in both leaves and corms of saffron. 81 % isolates showed lipase activity, 57 % cellulase, 48 % protease, 38 % amylase, 33 % chitinase and 29 % showed pectinase activity. 24 % of the isolates were phosphate solublizers, 86 % showed siderophore production and 80 % phytohormone production potential. The present repository of well characterized bacterial endophytes of saffron, have plant growth promoting potential which can be explored further for their respective roles in the biology of the saffron plant. PMID:26558164

  7. Control Efficacy of an Endophytic Bacillus amyloliquefaciens Strain BZ6-1 against Peanut Bacterial Wilt, Ralstonia solanacearum

    PubMed Central

    Liang, Guobin

    2014-01-01

    We aimed to isolate and identify endophytic bacteria that might have efficacy against peanut bacterial wilt (BW) caused by Ralstonia solanacearum. Thirty-seven endophytic strains were isolated from healthy peanut plants in R. solanacearum-infested fields and eight showed antagonistic effects against R. solanacearum. Strain BZ6-1 with the highest antimicrobial activity was identified as Bacillus amyloliquefaciens based on morphology, biochemistry, and 16S rRNA analysis. Culture conditions of BZ6-1 were optimized using orthogonal test method and inhibitory zone diameter in dual culture plate assay reached 34.2?mm. Furthermore, main antimicrobial substances of surfactin and fengycin A homologues produced by BZ6-1 were analyzed by high performance liquid chromatography electrospray ionization tandem mass spectrometry. Finally, pot experiments were adopted to test the control efficiency of BZ6-1 against peanut BW. Disease incidence decreased significantly from 84.5% in the control to 12.1% with addition of 15?mL (108 cfu?mL?1) culture broth for each seedling, suggesting the feasibility of strain BZ6-1 in the biological control of peanut plants BW. PMID:24527448

  8. Control efficacy of an endophytic Bacillus amyloliquefaciens strain BZ6-1 against peanut bacterial Wilt, Ralstonia solanacearum.

    PubMed

    Wang, Xiaobing; Liang, Guobin

    2014-01-01

    We aimed to isolate and identify endophytic bacteria that might have efficacy against peanut bacterial wilt (BW) caused by Ralstonia solanacearum. Thirty-seven endophytic strains were isolated from healthy peanut plants in R. solanacearum-infested fields and eight showed antagonistic effects against R. solanacearum. Strain BZ6-1 with the highest antimicrobial activity was identified as Bacillus amyloliquefaciens based on morphology, biochemistry, and 16S rRNA analysis. Culture conditions of BZ6-1 were optimized using orthogonal test method and inhibitory zone diameter in dual culture plate assay reached 34.2 mm. Furthermore, main antimicrobial substances of surfactin and fengycin A homologues produced by BZ6-1 were analyzed by high performance liquid chromatography electrospray ionization tandem mass spectrometry. Finally, pot experiments were adopted to test the control efficiency of BZ6-1 against peanut BW. Disease incidence decreased significantly from 84.5% in the control to 12.1% with addition of 15 mL (10(8) cfu mL(-1)) culture broth for each seedling, suggesting the feasibility of strain BZ6-1 in the biological control of peanut plants BW. PMID:24527448

  9. Characterization of endophytic strains of Bacillus mojavensis and their production of surfactin isomers

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Bacillus subtilis consists of a large collection of strains from which several cryptic species have been delineated, and most of these along with strains within the species are important biocontrol agents. Bacillus mojavensis, a species recently distinguished from this broad Bacillus subtilis grou...

  10. Endophyte-assisted promotion of biomass production and metal-uptake of energy crop sweet sorghum by plant-growth-promoting endophyte Bacillus sp. SLS18.

    PubMed

    Luo, Shenglian; Xu, Taoying; Chen, Liang; Chen, Jueliang; Rao, Chan; Xiao, Xiao; Wan, Yong; Zeng, Guangming; Long, Fei; Liu, Chengbin; Liu, Yutang

    2012-02-01

    The effects of Bacillus sp. SLS18, a plant-growth-promoting endophyte, on the biomass production and Mn/Cd uptake of sweet sorghum (Sorghum bicolor L.), Phytolacca acinosa Roxb., and Solanum nigrum L. were investigated. SLS18 displayed multiple heavy metals and antibiotics resistances. The strain also exhibited the capacity of producing indole-3-acetic acid, siderophores, and 1-aminocyclopropane-1-carboxylic acid deaminase. In pot experiments, SLS18 could not only infect plants effectively but also significantly increase the biomass of the three tested plants in the presence of Mn/Cd. The promoting effect order of SLS18 on the biomass of the tested plants was sweet sorghum > P. acinosa > S. nigrum L. In the presence of Mn (2,000 mg kg(-1)) and Cd (50 mg kg(-1)) in vermiculite, the total Mn/Cd uptakes in the aerial parts of sweet sorghum, P. acinosa, and S. nigrum L. were increased by 65.2%/40.0%, 55.2%/31.1%, and 18.6%/25.6%, respectively, compared to the uninoculated controls. This demonstrates that the symbiont of SLS18 and sweet sorghum has the potential of improving sweet sorghum biomass production and its total metal uptake on heavy metal-polluted marginal land. It offers the potential that heavy metal-polluted marginal land could be utilized in planting sweet sorghum as biofuel feedstock for ethanol production, which not only gives a promising phytoremediation strategy but also eases the competition for limited fertile farmland between energy crops and food crops. PMID:21792590

  11. Characterization of endophytic Bacillus strains from tomato plants (Lycopersicon esculentum) displaying antifungal activity against Botrytis cinerea Pers.

    PubMed

    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. PMID:26347324

  12. Endophytic Bacillus subtilis Strain E1R-J Is a Promising Biocontrol Agent for Wheat Powdery Mildew

    PubMed Central

    Gong, Yufei; Huo, Yunxia; Han, Qingmei; Kang, Zhensheng; Huang, Lili

    2015-01-01

    In this study, the biocontrol efficacies of 14 endophytic bacterial strains were tested against Blumeria graminis f. sp. tritici (Bgt) in pot experiments under greenhouse conditions. Bacillus subtilis strain E1R-j significantly reduced disease index and exhibited the best control (90.97%). When different formulations of E1R-j were sprayed 24?h before Bgt inoculation, fermentation liquid without bacterial cell and crude protein suspension displayed the similar effects; and they reduced disease index more than bacterial cell suspension (109?cfu?mL?1) and fermentation liquid without protein. The control effects were not significantly different between 1011 and 109?cfu?mL?1 of bacterial cell suspension but were higher than 107?cfu?mL?1. Further observations showed that conidial germination and appressorial formation of Bgt were retarded by spraying E1R-j 24?h before Bgt inoculation. Compared with the water check, conidial germination and appressorial formation were decreased by 43.3% and 42.7%, respectively. In the treatment with E1R-j, the number of houstoria significantly reduced and the speed of mycelial extension was slowed down in the wheat leaves. Scanning electron microscopy observation revealed that E1R-j significantly suppressed the conidial germination and caused rupture and deformation of germ tubes. On the surface of wheat leaves, mycelia and conidiophores became shrinking. PMID:25759819

  13. Lipopeptides from the Banyan Endophyte, Bacillus subtilis K1: Mass Spectrometric Characterization of a Library of Fengycins

    NASA Astrophysics Data System (ADS)

    Pathak, Khyati V.; Keharia, Haresh; Gupta, Kallol; Thakur, Suman S.; Balaram, Padmanabhan

    2012-10-01

    Mass spectrometric analysis of a banyan endophyte, Bacillus subtilis K1, extract showing broad spectrum antifungal activity revealed a complex mixture of lipopeptides, iturins, surfactins, and fengycins. Fractionation by reversed-phase high performance liquid chromatography (HPLC) facilitated a detailed analysis of fengycin microheterogeneity. Matrix assisted laser desorption ionization (MALDI) and electrospray ionization (ESI) mass spectrometric studies permitted the identification of several new fengycin variants. Four major sites of heterogeneity are identified: (1) N-terminus β-hydroxy fatty acid moiety, where chain length variation and the presence of unsaturation occur, (2) position 6 (Ala/Val/Ile/Leu), (3) position 10 (Val/Ile) within the macrocyclic ring, and (4) Gln to Glu replacement at position 8, resulting in fengycin variants that differ in mass by 1 Da. Diagnostic fragment ions provide a quick method for localizing the sites of variation in the macrocycle or the linear segment. Subsequent establishment of the sequences is achieved by MS/MS analysis of linear fengycin species produced by hydrolysis of the macrocyclic lactone. Unsaturation in the fatty acid chain and the presence of linear precursors in the B. subtilis K1 extract are also established by mass spectrometry. The anomalous distribution of intensities within isotopic multiplets is a diagnostic for Gln/Glu replacements. High resolution mass spectrometry facilitates the identification of fengycin species differing by 1 Da by localizing the variable position (Gln8/Glu8) in the fengycin variants.

  14. Biotechnological Potential of Agro Residues for Economical Production of Thermoalkali-Stable Pectinase by Bacillus pumilus dcsr1 by Solid-State Fermentation and Its Efficacy in the Treatment of Ramie Fibres.

    PubMed

    Sharma, Deepak Chand; Satyanarayana, T

    2012-01-01

    The production of a thermostable and highly alkaline pectinase by Bacillus pumilus dcsr1 was optimized in solid-state fermentation (SSF) and the impact of various treatments (chemical, enzymatic, and in combination) on the quality of ramie fibres was investigated. Maximum enzyme titer (348.0 ± 11.8 Ug(-1) DBB) in SSF was attained, when a mixture of agro-residues (sesame oilseed cake, wheat bran, and citrus pectin, 1 : 1 : 0.01) was moistened with mineral salt solution (a(w) 0.92, pH 9.0) at a substrate-to-moistening agent ratio of 1 : 2.5 and inoculated with 25% of 24 h old inoculum, in 144 h at 40°C. Parametric optimization in SSF resulted in 1.7-fold enhancement in the enzyme production as compared to that recorded in unoptimized conditions. A 14.2-fold higher enzyme production was attained in SSF as compared to that in submerged fermentation (SmF). The treatment with the enzyme significantly improved tensile strength and Young's modulus, reduction in brittleness, redness and yellowness, and increase in the strength and brightness of ramie fibres. PMID:22928091

  15. Biotechnological Potential of Agro Residues for Economical Production of Thermoalkali-Stable Pectinase by Bacillus pumilus dcsr1 by Solid-State Fermentation and Its Efficacy in the Treatment of Ramie Fibres

    PubMed Central

    Sharma, Deepak Chand; Satyanarayana, T.

    2012-01-01

    The production of a thermostable and highly alkaline pectinase by Bacillus pumilus dcsr1 was optimized in solid-state fermentation (SSF) and the impact of various treatments (chemical, enzymatic, and in combination) on the quality of ramie fibres was investigated. Maximum enzyme titer (348.0 ± 11.8 Ug−1 DBB) in SSF was attained, when a mixture of agro-residues (sesame oilseed cake, wheat bran, and citrus pectin, 1 : 1 : 0.01) was moistened with mineral salt solution (aw 0.92, pH 9.0) at a substrate-to-moistening agent ratio of 1 : 2.5 and inoculated with 25% of 24 h old inoculum, in 144 h at 40°C. Parametric optimization in SSF resulted in 1.7-fold enhancement in the enzyme production as compared to that recorded in unoptimized conditions. A 14.2-fold higher enzyme production was attained in SSF as compared to that in submerged fermentation (SmF). The treatment with the enzyme significantly improved tensile strength and Young's modulus, reduction in brittleness, redness and yellowness, and increase in the strength and brightness of ramie fibres. PMID:22928091

  16. Bacillus oryzicola sp. nov., an Endophytic Bacterium Isolated from the Roots of Rice with Antimicrobial, Plant Growth Promoting, and Systemic Resistance Inducing Activities in Rice.

    PubMed

    Chung, Eu Jin; Hossain, Mohammad Tofajjal; Khan, Ajmal; Kim, Kyung Hyun; Jeon, Che Ok; Chung, Young Ryun

    2015-06-01

    Biological control of major rice diseases has been attempted in several rice-growing countries in Asia during the last few decades and its application using antagonistic bacteria has proved to be somewhat successful for controlling various fungal diseases in field trials. Two novel endophytic Bacillus species, designated strains YC7007 and YC7010(T), with anti-microbial, plant growth-promoting, and systemic resistance-inducing activities were isolated from the roots of rice in paddy fields at Jinju, Korea, and their multifunctional activities were analyzed. Strain YC7007 inhibited mycelial growth of major rice fungal pathogens strongly in vitro. Bacterial blight and panicle blight caused by Xanthomonas oryzae pv. oryzae (KACC 10208) and Burkholderia glumae (KACC 44022), respectively, were also suppressed effectively by drenching a bacterial suspension (10(7) cfu/ml) of strain YC7007 on the rhizosphere of rice. Additionally, strain YC7007 promoted the growth of rice seedlings with higher germination rates and more tillers than the untreated control. The taxonomic position of the strains was also investigated. Phylogenetic analyses based on 16S rRNA gene sequences indicated that both strains belong to the genus Bacillus, with high similarity to the closely related strains, Bacillus siamensis KACC 15859(T) (99.67%), Bacillus methylotrophicus KACC 13105(T) (99.65%), Bacillus amyloliquefaciens subsp. plantarum KACC 17177(T) (99.60%), and Bacillus tequilensis KACC 15944(T) (99.45%). The DNA-DNA relatedness value between strain YC7010(T) and the most closely related strain, B. siamensis KACC 15859(T) was 50.4±3.5%, but it was 91.5±11.0% between two strains YC7007 and YC7010(T), indicating the same species. The major fatty acids of two strains were anteiso-C15:0 and iso C15:0. Both strains contained MK-7 as a major respiratory quinone system. The G+C contents of the genomic DNA of two strains were 50.5 mol% and 51.2 mol%, respectively. Based on these polyphasic studies, the two strains YC7007 and YC7010(T) represent novel species of the genus Bacillus, for which the name Bacillus oryzicola sp. nov. is proposed. The type strain is YC7010(T) (= KACC 18228(T)). Taken together, our findings suggest that novel endophytic Bacillus strains can be used for the biological control of rice diseases. PMID:26060434

  17. Bacillus oryzicola sp. nov., an Endophytic Bacterium Isolated from the Roots of Rice with Antimicrobial, Plant Growth Promoting, and Systemic Resistance Inducing Activities in Rice

    PubMed Central

    Chung, Eu Jin; Hossain, Mohammad Tofajjal; Khan, Ajmal; Kim, Kyung Hyun; Jeon, Che Ok; Chung, Young Ryun

    2015-01-01

    Biological control of major rice diseases has been attempted in several rice-growing countries in Asia during the last few decades and its application using antagonistic bacteria has proved to be somewhat successful for controlling various fungal diseases in field trials. Two novel endophytic Bacillus species, designated strains YC7007 and YC7010T, with anti-microbial, plant growth-promoting, and systemic resistance-inducing activities were isolated from the roots of rice in paddy fields at Jinju, Korea, and their multifunctional activities were analyzed. Strain YC7007 inhibited mycelial growth of major rice fungal pathogens strongly in vitro. Bacterial blight and panicle blight caused by Xanthomonas oryzae pv. oryzae (KACC 10208) and Burkholderia glumae (KACC 44022), respectively, were also suppressed effectively by drenching a bacterial suspension (107 cfu/ml) of strain YC7007 on the rhizosphere of rice. Additionally, strain YC7007 promoted the growth of rice seedlings with higher germination rates and more tillers than the untreated control. The taxonomic position of the strains was also investigated. Phylogenetic analyses based on 16S rRNA gene sequences indicated that both strains belong to the genus Bacillus, with high similarity to the closely related strains, Bacillus siamensis KACC 15859T (99.67%), Bacillus methylotrophicus KACC 13105T (99.65%), Bacillus amyloliquefaciens subsp. plantarum KACC 17177T (99.60%), and Bacillus tequilensis KACC 15944T (99.45%). The DNA-DNA relatedness value between strain YC7010T and the most closely related strain, B. siamensis KACC 15859T was 50.4±3.5%, but it was 91.5±11.0% between two strains YC7007 and YC7010T, indicating the same species. The major fatty acids of two strains were anteiso-C15:0 and iso C15:0. Both strains contained MK-7 as a major respiratory quinone system. The G+C contents of the genomic DNA of two strains were 50.5 mol% and 51.2 mol%, respectively. Based on these polyphasic studies, the two strains YC7007 and YC7010T represent novel species of the genus Bacillus, for which the name Bacillus oryzicola sp. nov. is proposed. The type strain is YC7010T (= KACC 18228T). Taken together, our findings suggest that novel endophytic Bacillus strains can be used for the biological control of rice diseases. PMID:26060434

  18. Screening and characterization of endophytic Bacillus and Paenibacillus strains from medicinal plant Lonicera japonica for use as potential plant growth promoters.

    PubMed

    Zhao, Longfei; Xu, Yajun; Lai, Xin-He; Shan, Changjuan; Deng, Zhenshan; Ji, Yuliang

    2015-12-01

    A total of 48 endophytic bacteria were isolated from surface-sterilized tissues of the medicinal plant Lonicera japonica, which is grown in eastern China; six strains were selected for further study based on their potential ability to promote plant growth in vitro (siderophore and indoleacetic acid production). The bacteria were characterized by phylogenetically analyzing their 16S rRNA gene similarity, by examining their effect on the mycelial development of pathogenic fungi, by testing their potential plant growth-promoting characteristics, and by measuring wheat growth parameters after inoculation. Results showed that the number of endophytic bacteria in L. japonica varied among different tissues, but it remained relatively stable in the same tissues from four different plantation locations. Among the three endophytic strains, strains 122 and 124 both had high siderophore production, with the latter showing the highest phosphate solubilization activity (45.6 mg/L) and aminocyclopropane-1-carboxylic acid deaminase activity (47.3 nmol/mg/h). Strain 170 had the highest indoleacetic acid (IAA) production (49.2 mg/L) and cellulase and pectinase activities. After inoculation, most of the six selected isolates showed a strong capacity to promote wheat growth. Compared with the controls, the increase in the shoot length, root length, fresh weight, dry weight, and chlorophyll content was most remarkable in wheat seedlings inoculated with strain 130. The positive correlation between enzyme (cellulose and pectinase) activity and inhibition rate on Fusarium oxysporum, the IAA production, and the root length of wheat seedlings inoculated with each tested endophytic strain was significant in regression analysis. Deformity of pathogenic fungal mycelia was observed under a microscope after the interaction with the endophytic isolates. Such deformity may be directly related to the production of hydrolytic bacterial enzymes (cellulose and pectinase). The six endophytic bacterial strains were identified to be Paenibacillus and Bacillus strains based on the results of 16S rRNA gene sequencing analysis and their physiological and biochemical characteristics. Results indicate the promising application of endophytic bacteria to the biological control of pathogenic fungi and the improvement of wheat crop growth. PMID:26691455

  19. Screening and characterization of endophytic Bacillus and Paenibacillus strains from medicinal plant Lonicera japonica for use as potential plant growth promoters

    PubMed Central

    Zhao, Longfei; Xu, Yajun; Lai, Xin-He; Shan, Changjuan; Deng, Zhenshan; Ji, Yuliang

    2015-01-01

    Abstract A total of 48 endophytic bacteria were isolated from surface-sterilized tissues of the medicinal plant Lonicera japonica, which is grown in eastern China; six strains were selected for further study based on their potential ability to promote plant growth in vitro (siderophore and indoleacetic acid production). The bacteria were characterized by phylogenetically analyzing their 16S rRNA gene similarity, by examining their effect on the mycelial development of pathogenic fungi, by testing their potential plant growth-promoting characteristics, and by measuring wheat growth parameters after inoculation. Results showed that the number of endophytic bacteria in L. japonica varied among different tissues, but it remained relatively stable in the same tissues from four different plantation locations. Among the three endophytic strains, strains 122 and 124 both had high siderophore production, with the latter showing the highest phosphate solubilization activity (45.6 mg/L) and aminocyclopropane-1-carboxylic acid deaminase activity (47.3 nmol/mg/h). Strain 170 had the highest indoleacetic acid (IAA) production (49.2 mg/L) and cellulase and pectinase activities. After inoculation, most of the six selected isolates showed a strong capacity to promote wheat growth. Compared with the controls, the increase in the shoot length, root length, fresh weight, dry weight, and chlorophyll content was most remarkable in wheat seedlings inoculated with strain 130. The positive correlation between enzyme (cellulose and pectinase) activity and inhibition rate on Fusarium oxysporum, the IAA production, and the root length of wheat seedlings inoculated with each tested endophytic strain was significant in regression analysis. Deformity of pathogenic fungal mycelia was observed under a microscope after the interaction with the endophytic isolates. Such deformity may be directly related to the production of hydrolytic bacterial enzymes (cellulose and pectinase). The six endophytic bacterial strains were identified to be Paenibacillus and Bacillus strains based on the results of 16S rRNA gene sequencing analysis and their physiological and biochemical characteristics. Results indicate the promising application of endophytic bacteria to the biological control of pathogenic fungi and the improvement of wheat crop growth. PMID:26691455

  20. Bacillus endoradicis sp. nov., an endophytic bacterium isolated from soybean root.

    PubMed

    Zhang, Yun Zeng; Chen, Wen Feng; Li, Mao; Sui, Xin Hua; Liu, Hong-Can; Zhang, Xiao Xia; Chen, Wen Xin

    2012-02-01

    A gram-positive, aerobic, motile rod, designated strain CCBAU 05776(T), was isolated from the inner tissues of a healthy soybean (Glycine max L.) root collected from an agricultural field in the countryside of Shijiazhuang city, Hebei Province, China. Phylogenetic analysis of the 16S rRNA gene indicated that this strain was most closely related to Bacillus muralis LMG 20238(T) and Bacillus simplex NBRC 15720(T) with similarity of 96.5?% and 96.3?%, respectively, lower than the suggested threshold (97.0?%) for separating bacterial species. In phenotypic characterization, the novel strain differed from the two most related species in that it did not hydrolyse casein or starch but could grow on MacConkey agar. It grew between 15 and 45 °C and tolerated up to 7?% NaCl (w/v). Strain CCBAU 05776(T) grew in media with pH 5.5 to 10 (optimal growth at pH 7.0-8.0). The predominant cellular fatty acids were iso-C(15?:?0) (40.81?%) and C(16?:?1)?7c alcohol (10.61?%). The predominant isoprenoid quinone was menaquinone 7 (MK-7). The cell-wall peptidoglycan contained meso-diaminopimelic acid. The major polar lipids were diphosphatidylglycerol and phosphatidylglycerol. The DNA G+C was 40.8 mol% (T(m)). DNA-DNA relatedness of the novel isolate with B. muralis and B. simplex was 42.4?% and 32.7?%, respectively. Based upon the consensus of phylogenetic and phenotypic analyses, strain CCBAU 05776(T) represents a novel species within the genus Bacillus, for which the name Bacillus endoradicis sp. nov. is proposed. The type strain is CCBAU 05776(T) (?=?LMG 25492(T) ?=?HAMBI 3097(T)). PMID:21441377

  1. The hyperaccumulator Sedum plumbizincicola harbors metal-resistant endophytic bacteria that improve its phytoextraction capacity in multi-metal contaminated soil.

    PubMed

    Ma, Ying; Oliveira, Rui S; Nai, Fengjiao; Rajkumar, Mani; Luo, Yongming; Rocha, Inês; Freitas, Helena

    2015-06-01

    Endophyte-assisted phytoremediation has recently been suggested as a successful approach for ecological restoration of metal contaminated soils, however little information is available on the influence of endophytic bacteria on the phytoextraction capacity of metal hyperaccumulating plants in multi-metal polluted soils. The aims of our study were to isolate and characterize metal-resistant and 1-aminocyclopropane-1-carboxylate (ACC) utilizing endophytic bacteria from tissues of the newly discovered Zn/Cd hyperaccumulator Sedum plumbizincicola and to examine if these endophytic bacterial strains could improve the efficiency of phytoextraction of multi-metal contaminated soils. Among a collection of 42 metal resistant bacterial strains isolated from the tissues of S. plumbizincicola grown on Pb/Zn mine tailings, five plant growth promoting endophytic bacterial strains (PGPE) were selected due to their ability to promote plant growth and to utilize ACC as the sole nitrogen source. The five isolates were identified as Bacillus pumilus E2S2, Bacillus sp. E1S2, Bacillus sp. E4S1, Achromobacter sp. E4L5 and Stenotrophomonas sp. E1L and subsequent testing revealed that they all exhibited traits associated with plant growth promotion, such as production of indole-3-acetic acid and siderophores and solubilization of phosphorus. These five strains showed high resistance to heavy metals (Cd, Zn and Pb) and various antibiotics. Further, inoculation of these ACC utilizing strains significantly increased the concentrations of water extractable Cd and Zn in soil. Moreover, a pot experiment was conducted to elucidate the effects of inoculating metal-resistant ACC utilizing strains on the growth of S. plumbizincicola and its uptake of Cd, Zn and Pb in multi-metal contaminated soils. Out of the five strains, B. pumilus E2S2 significantly increased root (146%) and shoot (17%) length, fresh (37%) and dry biomass (32%) of S. plumbizincicola as well as plant Cd uptake (43%), whereas Bacillus sp. E1S2 significantly enhanced the accumulation of Zn (18%) in plants compared with non-inoculated controls. The inoculated strains also showed high levels of colonization in rhizosphere and plant tissues. Results demonstrate the potential to improve phytoextraction of soils contaminated with multiple heavy metals by inoculating metal hyperaccumulating plants with their own selected functional endophytic bacterial strains. PMID:25796039

  2. Unprecedented access of phenolic substrates to the heme active site of a catalase: substrate binding and peroxidase-like reactivity of Bacillus pumilus catalase monitored by X-ray crystallography and EPR spectroscopy.

    PubMed

    Loewen, Peter C; Villanueva, Jacylyn; Switala, Jacek; Donald, Lynda J; Ivancich, Anabella

    2015-05-01

    Heme-containing catalases and catalase-peroxidases catalyze the dismutation of hydrogen peroxide as their predominant catalytic activity, but in addition, individual enzymes support low levels of peroxidase and oxidase activities, produce superoxide, and activate isoniazid as an antitubercular drug. The recent report of a heme enzyme with catalase, peroxidase and penicillin oxidase activities in Bacillus pumilus and its categorization as an unusual catalase-peroxidase led us to investigate the enzyme for comparison with other catalase-peroxidases, catalases, and peroxidases. Characterization revealed a typical homotetrameric catalase with one pentacoordinated heme b per subunit (Tyr340 being the axial ligand), albeit in two orientations, and a very fast catalatic turnover rate (kcat ?=?339,000 s(-1) ). In addition, the enzyme supported a much slower (kcat ?=?20 s(-1) ) peroxidatic activity utilizing substrates as diverse as ABTS and polyphenols, but no oxidase activity. Two binding sites, one in the main access channel and the other on the protein surface, accommodating pyrogallol, catechol, resorcinol, guaiacol, hydroquinone, and 2-chlorophenol were identified in crystal structures at 1.65-1.95 Å. A third site, in the heme distal side, accommodating only pyrogallol and catechol, interacting with the heme iron and the catalytic His and Arg residues, was also identified. This site was confirmed in solution by EPR spectroscopy characterization, which also showed that the phenolic oxygen was not directly coordinated to the heme iron (no low-spin conversion of the Fe(III) high-spin EPR signal upon substrate binding). This is the first demonstration of phenolic substrates directly accessing the heme distal side of a catalase. PMID:25663126

  3. Bacillus mojavensis: its endophytic nature, the surfactins and their role in the plant response to infection by Fusarium verticillioides

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Bacterial endophytes are fundamentally important as natural components of most plants, wild and cultivated with strong ecological merits. These ancient associations are recently at the forefront of biological control strategies designed to circumvent the problems associated with pesticide uses, par...

  4. Enhanced degradation of textile effluent in constructed wetland system using Typha domingensis and textile effluent-degrading endophytic bacteria.

    PubMed

    Shehzadi, Maryam; Afzal, Muhammad; Khan, Muhammad Umar; Islam, Ejazul; Mobin, Amina; Anwar, Samina; Khan, Qaiser Mahmood

    2014-07-01

    Textile effluent is one of the main contributors of water pollution and it adversely affects fauna and flora. Constructed wetland is a promising approach to remediate the industrial effluent. The detoxification of industrial effluent in a constructed wetland system may be enhanced by applying beneficial bacteria that are able to degrade contaminants present in industrial effluent. The aim of this study was to evaluate the influence of inoculation of textile effluent-degrading endophytic bacteria on the detoxification of textile effluent in a vertical flow constructed wetland reactor. A wetland plant, Typha domingensis, was vegetated in reactor and inoculated with two endophytic bacterial strains, Microbacterium arborescens TYSI04 and Bacillus pumilus PIRI30. These strains possessed textile effluent-degrading and plant growth-promoting activities. Results indicated that bacterial inoculation improved plant growth, textile effluent degradation and mutagenicity reduction and were correlated with the population of textile effluent-degrading bacteria in the rhizosphere and endosphere of T. domingensis. Bacterial inoculation enhanced textile effluent-degrading bacterial population in rhizosphere, root and shoot of T. domingensis. Significant reductions in COD (79%), BOD (77%) TDS (59%) and TSS (27%) were observed by the combined use of plants and bacteria within 72 h. The resultant effluent meets the wastewater discharge standards of Pakistan and can be discharged into the environment without any risks. This study revealed that the combined use of plant and endophytic bacteria is one of the approaches to enhance textile effluent degradation in a constructed wetland system. PMID:24755300

  5. Purification, characterization, and heterologous expression of an antifungal protein from the endophytic Bacillus subtilis strain Em7 and its activity against Sclerotinia sclerotiorum.

    PubMed

    Wang, N N; Gao, X N; Yan, X; Li, Z P; Kang, Z S; Huang, L L; Han, Q M

    2015-01-01

    An antifungal protein exhibiting a high activity against Sclero-tinia sclerotiorum in vivo was purified by ammonium sulfate precipitation, hydrophobic chromatography, and gel filtration chromatography from the culture filtrate of the endophytic Bacillus subtilis strain Em7. The protein was characterized as a β-1,3-1,4-glucanase according to amino acid analysis, and showed excellent properties in thermal stability and acid resistance. At the same time, the antifungal protein was cloned and het-erologously expressed in Escherichia coli BL21. The recombinant protein was purified and showed similar enzymatic properties to the native protein, exhibiting strong inhibitory activity against S. sclerotiorum. This shows that the β-1,3-1,4-glucanase may play a very important role in B. subtilis Em7 biocontrol function. In addition, many physiochemical properties of the na-tive and purified recombinant protein were compared, including the effect of pH, temperature, metal cations, substrate specificity, and kinetic param-eters. All parameters were similar between the native and recombinant pu-rified protein, indicating that the purified recombinant protein has potential for industrial applications. PMID:26634515

  6. Induction of Defense-Related Ultrastructural Modifications in Pea Root Tissues Inoculated with Endophytic Bacteria.

    PubMed Central

    Benhamou, N.; Kloepper, J. W.; Quadt-Hallman, A.; Tuzun, S.

    1996-01-01

    The stimulation exerted by the endophytic bacterium Bacillus pumilus strain SE34 in plant defense reactions was investigated at the ultrastructural level using an in vitro system in which root-inducing T-DNA pea (Pisum sativum L.) roots were infected with the pea root-rotting fungus Fusarium oxysporum f. sp. pisi. In nonbacterized roots, the pathogen multiplied abundantly through much of the tissue including the vascular stele, whereas in prebacterized roots, pathogen growth was restricted to the epidermis and the outer cortex In these prebacterized roots, typical host reactions included strengthening the epidermal and cortical cell walls and deposition of newly formed barriers beyond the infection sites. Wall appositions were found to contain large amounts of callose in addition to being infiltrated with phenolic compounds. The labeling pattern obtained with the gold-complexed laccase showed that phenolics were widely distributed in Fusarium-challenged, bacterized roots. Such compounds accumulated in the host cell walls and the intercellular spaces as well as at the surface or even inside of the invading hyphae of the pathogen. The wall-bound chitin component in Fusarium hyphae colonizing bacterized roots was preserved even when hyphae had undergone substantial degradation. These observations confirm that endophytic bacteria may function as potential inducers of plant disease resistance. PMID:12226427

  7. Bacterial endophytes: Bacillus spp. from vegetable crops as potential biological control agents of black pod rot of cacao

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Diseases are the most important factors limiting the production of Theobroma cacao in South America. Because of high disease pressure and environmental concerns, biological control is a pertinent area of research for cacao disease management. In this work, we evaluated the ability of four Bacillus s...

  8. Purification and characterization of a highly active chromate reductase from endophytic Bacillus sp. DGV19 of Albizzia lebbeck (L.) Benth. actively involved in phytoremediation of tannery effluent-contaminated sites.

    PubMed

    Manikandan, Muthu; Gopal, Judy; Kumaran, Rangarajulu Senthil; Kannan, Vijayaraghavan; Chun, Sechul

    2016-02-17

    Phytoremediation using timber-yielding tree species is considered to be the most efficient method for chromium/tannery effluent-contaminated sites. In this study, we have chosen Albizzia lebbeck, a chromium hyperaccumulator plant, and studied one of its chromium detoxification processes operated by its endophytic bacterial assemblage. Out of the four different groups of endophytic bacteria comprising Pseudomonas, Rhizobium, Bacillus, and Salinicoccus identified from A. lebbeck employed in phytoremediation of tannery effluent-contaminated soil, Bacillus predominated with three species, which exhibited not only remarkable chromium accumulation ability but also high chromium reductase activity. A chromate reductase was purified to homogeneity from the most efficient chromium accumulator, Bacillus sp. DGV 019, and the purified 34.2-kD enzyme was observed to be stable at temperatures from 20°C to 60°C. The enzyme was active over a wide range of pH values (4.0-9.0). Furthermore, the enzyme activity was enhanced with the electron donors NADH, followed by NADPH, not affected by glutathione and ascorbic acid. Cu(2+) enhanced the activity of the purified enzyme but was inhibited by Zn(2+) and etheylenediamine tetraacetic acid (EDTA). In conclusion, due to its versatile adaptability the chromate reductase can be used for chromium remediation. PMID:26444299

  9. Inoculation with endophytic Bacillus megaterium 1Y31 increases Mn accumulation and induces the growth and energy metabolism-related differentially-expressed proteome in Mn hyperaccumulator hybrid pennisetum.

    PubMed

    Zhang, Wen-Hui; He, Lin-Yan; Wang, Qi; Sheng, Xia-Fang

    2015-12-30

    In this study, a hydroponic culture experiment was conducted in a greenhouse to investigate the molecular and microbial mechanisms involved in the endophytic Bacillus megaterium 1Y31-enhanced Mn tolerance and accumulation in Mn hyperaccumulator hybrid pennisetum. Strain 1Y31 significantly increased the dry weights (ranging from 28% to 94%) and total Mn uptake (ranging from 23% to 112%) of hybrid pennisetum treated with 0, 2, and 10mM Mn compared to the control. Total 98 leaf differentially expressed proteins were identified between the live and dead bacterial inoculated hybrid pennisetum. The major leaf differentially expressed proteins were involved in energy generation, photosynthesis, response to stimulus, metabolisms, and unknown function. Furthermore, most of the energy generation and photosynthesis-related proteins were up-regulated, whereas most of the response to stimulus and metabolism-related proteins were down-regulated under Mn stress. Notably, the proportion of indole-3-acetic acid (IAA)-producing endophytic bacteria was significantly higher in the bacterial inoculated plants under Mn stress. The results suggested that strain 1Y31 increased the growth and Mn uptake of hybrid pennisetum through increasing the efficiency of photosynthesis and energy metabolism as well as the proportion of plant growth-promoting endophytic bacteria in the plants. PMID:26241871

  10. COFFEE ENDOPHYTES

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A survey for fungal endophytes in various coffee (Coffea arabica L.) tissues was conducted in Colombia, Hawaii, Mexico, and Puerto Rico. More than 700 fungal endophyte isolates have been sequenced: 281 from Colombia, 240 from Hawaii, 119 from Mexico, and 68 from Puerto Rico; these comprise over 17...

  11. Inoculation of Phaseolus vulgaris with the nodule-endophyte Agrobacterium sp. 10C2 affects richness and structure of rhizosphere bacterial communities and enhances nodulation and growth.

    PubMed

    Chihaoui, Saif-Allah; Trabelsi, Darine; Jdey, Ahmed; Mhadhbi, Haythem; Mhamdi, Ridha

    2015-08-01

    Agrobacterium sp. 10C2 is a nonpathogenic and non-symbiotic nodule-endophyte strain isolated from root nodules of Phaseolus vulgaris. The effect of this strain on nodulation, plant growth and rhizosphere bacterial communities of P. vulgaris is investigated under seminatural conditions. Inoculation with strain 10C2 induced an increase in nodule number (+54 %) and plant biomass (+16 %). Grains also showed a significant increase in phosphorus (+53 %), polyphenols (+217 %), flavonoids (+62 %) and total antioxidant capacity (+82 %). The effect of strain 10C2 on bacterial communities was monitored using terminal restriction fragment length polymorphism of PCR-amplified 16S rRNA genes. When the initial soil was inoculated with strain 10C2 and left 15 days, the Agrobacterium strain did not affect TRF richness but changed structure. When common bean was sown in these soils and cultivated during 75 days, both TRF richness and structure were affected by strain 10C2. TRF richness increased in the rhizosphere soil, while it decreased in the bulk soil (root free). The taxonomic assignation of TRFs induced by strain 10C2 in the bean rhizosphere revealed the presence of four phyla (Firmicutes, Actinobacteria, Bacteroidetes and Proteobacteria) with a relative preponderance of Firmicutes, represented mainly by Bacillus species. Some of these taxa (i.e., Bacillus licheniformis, Bacillus pumilus, Bacillus senegalensis, Bacillus subtilis, Bacillus firmus and Paenibacillus koreensis) are particularly known for their plant growth-promoting potentialities. These results suggest that the beneficial effects of strain 10C2 observed on plant growth and grain quality are explained at least in part by the indirect effect through the promotion of beneficial microorganisms. PMID:25967041

  12. Microbial genotyping and differentiating between Bacillus mojavensis and Bacillus subtilis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Bacillus mojavensis, a specie recently distinguished from its previous Bacillus subtilis classification, was discovered in corn kernels and later determined to possess endophytic character. The bacterium was also determined to have biocontrol potential due to its growth inhibition of the maize mycot...

  13. The characterization and diversity of bacterial endophytes of grapevine.

    PubMed

    West, E R; Cother, E J; Steel, C C; Ash, G J

    2010-03-01

    The diversity of culturable and nonculturable bacterial endophytes of grapevine (Vitis vinifera L.) was examined using a combination of cultivation and molecular methods. Entire grapevines were sampled to characterize bacterial diversity from different locations throughout the vine. Gas chromatography of fatty acid methyl esters (FAMEs) was used to identify culturable isolates prior to subsequent further microbiological characterization, whilst denaturing gradient gel electrophoresis (DGGE) was used to profile the ribosomal DNA of the bacterial endophyte community extracted from grapevines. Gas chromatography of FAMEs identified 75% of culturable bacterial endophytes to genus level (similarity index >0.3). Many isolates were identified as Bacillus spp., Pseudomonas spp., and Curtobacterium spp. Additionally, actinomycetes are reported for the first time as endophytes of grapevines, with a number of isolates identified as Streptomyces spp. DGGE was successfully used to identify major bands present in samples and indicated a degree of homogeneity of bacterial endophyte community profiles within the grapevines sampled. The major bacterial bands were sequenced and used in identification. Comparison with bacterial markers produced from cultured bacterial endophytes suggested that bacteria in the DGGE profiles were not the species most commonly cultured. Additional research demonstrated similarities between epiphytic and endophytic populations and examined potential entry vectors. Endophyte entry was demonstrated in both field-grown and potted grapevines ('Chardonnay') using a rifampicin-resistant Bacillus cereus mutant. The possibility of grapevine epiphytes becoming endophytes, if the opportunity arises, was supported by comparison of gas chromatography of FAMEs from epiphytic and endophytic populations. This research adds grapevine bacterial endophyte communities to those that have been characterized by a multifaceted approach. PMID:20453907

  14. Endophytic bacteria improve seedling growth of sunflower under water stress, produce salicylic acid, and inhibit growth of pathogenic fungi.

    PubMed

    Forchetti, Gabriela; Masciarelli, Oscar; Izaguirre, María J; Alemano, Sergio; Alvarez, Daniel; Abdala, Guillermina

    2010-12-01

    Endophytic bacterial strains SF2 (99.9% homology with Achromobacter xylosoxidans), and SF3 and SF4 (99.9% homology with Bacillus pumilus) isolated from sunflower grown under irrigation or drought were selected on the basis of plant growth-promoting bacteria (PGPB) characteristics. Aims of the study were to examine effects of inoculation with SF2, SF3, and SF4 on sunflower cultivated under water stress, to evaluate salicylic acid (SA) production by these strains in control medium or at ?a = -2.03 MPa, and to analyze effects of exogenously applied SA, jasmonic acid (JA), bacterial pellets, and bacterial supernatants on growth of pathogenic fungi Alternaria sp., Sclerotinia sp., and Verticillum sp. Growth response to bacterial inoculation was studied in two inbred lines (water stress-sensitive B59 and water stress-tolerant B71) and commercial hybrid Paraiso 24. Under both water stress and normal conditions, plant growth following inoculation was more strongly enhanced for Paraiso 24 and B71 than for B59. All three strains produced SA in control medium; levels for SF3 and SF4 were higher than for SF2. SA production was dramatically higher at ?a = -2.03 MPa. Exogenously applied SA or JA caused a significant reduction of growth for Sclerotinia and a lesser reduction for Alternaria and Verticillum. Fungal growth was more strongly inhibited by bacterial pellets than by bacterial supernatants. Our findings indicate that these endophytic bacteria enhance growth of sunflower seedlings under water stress, produce SA, and inhibit growth of pathogenic fungi. These characteristics are useful for formulation of inoculants to improve growth and yield of sunflower crops. PMID:20383767

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

    PubMed Central

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

    2011-01-01

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

  16. Nitrogen acquisition in Agave tequilana from degradation of endophytic bacteria

    PubMed Central

    Beltran-Garcia, Miguel J.; White, Jr., James F.; Prado, Fernanda M.; Prieto, Katia R.; Yamaguchi, Lydia F.; Torres, Monica S.; Kato, Massuo J.; Medeiros, Marisa H. G.; Di Mascio, Paolo

    2014-01-01

    Plants form symbiotic associations with endophytic bacteria within tissues of leaves, stems, and roots. It is unclear whether or how plants obtain nitrogen from these endophytic bacteria. Here we present evidence showing nitrogen flow from endophytic bacteria to plants in a process that appears to involve oxidative degradation of bacteria. In our experiments we employed Agave tequilana and its seed-transmitted endophyte Bacillus tequilensis to elucidate organic nitrogen transfer from 15N-labeled bacteria to plants. Bacillus tequilensis cells grown in a minimal medium with 15NH4Cl as the nitrogen source were watered onto plants growing in sand. We traced incorporation of 15N into tryptophan, deoxynucleosides and pheophytin derived from chlorophyll a. Probes for hydrogen peroxide show its presence during degradation of bacteria in plant tissues, supporting involvement of reactive oxygen in the degradation process. In another experiment to assess nitrogen absorbed as a result of endophytic colonization of plants we demonstrated that endophytic bacteria potentially transfer more nitrogen to plants and stimulate greater biomass in plants than heat-killed bacteria that do not colonize plants but instead degrade in the soil. Findings presented here support the hypothesis that some plants under nutrient limitation may degrade and obtain nitrogen from endophytic microbes. PMID:25374146

  17. Distinct differentiation of closely related species of Bacillus subtilis group with industrial importance.

    PubMed

    Jeyaram, Kumaraswamy; Romi, Wahengbam; Singh, Thangjam Anand; Adewumi, Gbenga Adedeji; Basanti, Khundrakpam; Oguntoyinbo, Folarin Anthony

    2011-11-01

    PCR amplification of 16S rRNA gene by universal primers followed by restriction fragment length polymorphism analysis using RsaI, CfoI and HinfI endonucleases, distinctly differentiated closely related Bacillus amyloliquefaciens, Bacillus licheniformis and Bacillus pumilus from Bacillus subtilis sensu stricto. This simple, economical, rapid and reliable protocol could be an alternative to misleading phenotype-based grouping of these closely related species. PMID:21889958

  18. Isolation and characterization of bacterial endophytes of Chelidonium majus L.

    PubMed

    Goryluk, Agata; Rekosz-Burlaga, Hanna; Błaszczyk, Mieczysław

    2009-01-01

    The aim of this study was to isolate and identify endophytic bacteria from stems of Chelidonium majus L. (greater celandine) and to evaluate their antifungal properties. In total, 34 bacterial endophyte strains were isolated. The fungistatic effects of these bacteria on the growth of five moulds (Alternaria alternata, Chaetonium sp., Paecilomyces variotti, Byssochlamys fulva, Aureobasidium pullulans) and one species of black yeast (Exophiala mesophila) were tested. The majority of the bacterial isolates were found to inhibit the growth of fungi and those with the strongest antifungal properties were further characterized. Of the twelve isolates examined, 11 were species of Bacillus thuringiensis and one was Bacillus amyloliquefaciens. PMID:20380146

  19. Search for endophytic diazotrophs in barley seeds

    PubMed Central

    Zawoznik, Myriam S.; Vázquez, Susana C.; Díaz Herrera, Silvana M.; Groppa, María D.

    2014-01-01

    Eight endophytic isolates assigned to Pseudomonas, Azospirillum, and Bacillus genera according to pheno-genotypic features were retrieved from barley seeds under selective pressure for nitrogen-fixers. Genetic relationships among related isolates were investigated through RAPD. Six isolates displayed nitrogen-fixing ability, while all could biosynthesize indolacetic acid in vitro and showed no antibiosis effects against Azospirillum brasilense Az39, a recognized PGPR. PMID:25242949

  20. Electrotransformation of Bacillus mojavensis with fluorescent protein markers

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Gram-positive endophytic bacteria are difficult to transform. To study endophytic interactions between Bacillus mojavensis and maize, a method was developed to transform this species by electroporation with three fluorescent protein expressing integrative plasmids: pSG1154, pSG1192, and pSG1193. The...

  1. Diversity and potential application of endophytic bacteria in ginger.

    PubMed

    Chen, T; Chen, Z; Ma, G H; Du, B H; Shen, B; Ding, Y Q; Xu, K

    2014-01-01

    Here, 248 endophytic bacterial strains were isolated to assess the distribution and population diversity of endophytic bacteria in ginger plants. A total of 10.4 x 10(4) to 20.2 x 10(4) CFU/g fresh weight endophytic bacteria of different growth stages were isolated. Maximum bacterium numbers were obtained at the seedling stage. A total of 107 functional strains were screened, including 17 antibacterial strains and 90 indole acetic acid-producing strains. Based on 16S rDNA sequence restriction fragment length polymorphism and 16S rDNA sequences, these 107 strains were mapped and grouped into 16 genera. Bacillus and Pseudomonas were the dominant genera; however, the bacteria belonged to a tremendous range of genera, with the highest species richness being observed at the seedling stage. Sixteen strains exhibited antimicrobial activity against Pythium myriotylum Drechsler, while 7 strains exhibited antimicrobial activity against Phyllosticta zingiberi Hori. Bacillus was the dominant antibacterial strain. Pseudomonas fluorescens, B. megaterium, and Enterobacter ludwigii produced remarkably high levels of IAA. Only a few endophytic bacterial strains were inhibited in fresh ginger juice. Most of these strains were present during seedling stage, including Roseateles depolymerans, Chryseobacterium taiwanense, E. ludwigii, Agrobacterium larrymoorei, P. fluorescens, and Bacillus amyloliquefaciens. This study indicates that the community of endophytic bacteria in ginger changes with the synthesis of antibacterial substances. PMID:25062479

  2. Seasonal variation of bacterial endophytes in urban trees

    PubMed Central

    Shen, Shu Yi; Fulthorpe, Roberta

    2015-01-01

    Bacterial endophytes, non-pathogenic bacteria residing within plants, contribute to the growth and development of plants and their ability to adapt to adverse conditions. In order to fully exploit the capabilities of these bacteria, it is necessary to understand the extent to which endophytic communities vary between species and over time. The endophytes of Acer negundo, Ulmus pumila, and Ulmus parvifolia were sampled over three seasons and analyzed using culture dependent and independent methods (culture on two media, terminal restriction fragment length polymorphism, and tagged pyrosequencing of 16S ribosomal amplicons). The majority of culturable endophytes isolated were Actinobacteria, and all the samples harbored Bacillus, Curtobacterium, Frigoribacterium, Methylobacterium, Paenibacilllus, and Sphingomonas species. Regardless of culture medium used, only the culturable communities obtained in the winter for A. negundo could be distinguished from those of Ulmus spp. In contrast, the nonculturable communities were dominated by Proteobacteria and Actinobacteria, particularly Erwinia, Ralstonia, and Sanguibacter spp. The presence and abundance of various bacterial classes and phyla changed with the changing seasons. Multivariate analysis on the culture independent data revealed significant community differences between the endophytic communities of A. negundo and Ulmus spp., but overall season was the main determinant of endophytic community structure. This study suggests studies on endophytic populations of urban trees should expect to find significant seasonal and species-specific community differences and sampling should proceed accordingly. PMID:26042095

  3. In Vitro Assessment of Marine Bacillus for Use as Livestock Probiotics

    PubMed Central

    Prieto, Maria Luz; O’Sullivan, Laurie; Tan, Shiau Pin; McLoughlin, Peter; Hughes, Helen; Gutierrez, Montserrat; Lane, Jonathan A.; Hickey, Rita M.; Lawlor, Peadar G.; Gardiner, Gillian E.

    2014-01-01

    Six antimicrobial-producing seaweed-derived Bacillus strains were evaluated in vitro as animal probiotics, in comparison to two Bacillus from an EU-authorized animal probiotic product. Antimicrobial activity was demonstrated on solid media against porcine Salmonella and E. coli. The marine isolates were most active against the latter, had better activity than the commercial probiotics and Bacillus pumilus WIT 588 also reduced E. coli counts in broth. All of the marine Bacillus tolerated physiological concentrations of bile, with some as tolerant as one of the probiotics. Spore counts for all isolates remained almost constant during incubation in simulated gastric and ileum juices. All of the marine Bacillus grew anaerobically and the spores of all except one isolate germinated under anaerobic conditions. All were sensitive to a panel of antibiotics and none harbored Bacillus enterotoxin genes but all, except B. pumilus WIT 588, showed some degree of β-hemolysis. However, trypan blue dye exclusion and xCELLigence assays demonstrated a lack of toxicity in comparison to two pathogens; in fact, the commercial probiotics appeared more cytotoxic than the majority of the marine Bacillus. Overall, some of the marine-derived Bacillus, in particular B. pumilus WIT 588, demonstrate potential for use as livestock probiotics. PMID:24796302

  4. In vitro assessment of marine Bacillus for use as livestock probiotics.

    PubMed

    Prieto, Maria Luz; O'Sullivan, Laurie; Tan, Shiau Pin; McLoughlin, Peter; Hughes, Helen; Gutierrez, Montserrat; Lane, Jonathan A; Hickey, Rita M; Lawlor, Peadar G; Gardiner, Gillian E

    2014-05-01

    Six antimicrobial-producing seaweed-derived Bacillus strains were evaluated in vitro as animal probiotics, in comparison to two Bacillus from an EU-authorized animal probiotic product. Antimicrobial activity was demonstrated on solid media against porcine Salmonella and E. coli. The marine isolates were most active against the latter, had better activity than the commercial probiotics and Bacillus pumilus WIT 588 also reduced E. coli counts in broth. All of the marine Bacillus tolerated physiological concentrations of bile, with some as tolerant as one of the probiotics. Spore counts for all isolates remained almost constant during incubation in simulated gastric and ileum juices. All of the marine Bacillus grew anaerobically and the spores of all except one isolate germinated under anaerobic conditions. All were sensitive to a panel of antibiotics and none harbored Bacillus enterotoxin genes but all, except B. pumilus WIT 588, showed some degree of β-hemolysis. However, trypan blue dye exclusion and xCELLigence assays demonstrated a lack of toxicity in comparison to two pathogens; in fact, the commercial probiotics appeared more cytotoxic than the majority of the marine Bacillus. Overall, some of the marine-derived Bacillus, in particular B. pumilus WIT 588, demonstrate potential for use as livestock probiotics. PMID:24796302

  5. The status of the species Bacillus aerophilus and Bacillus stratosphericus. Request for an Opinion.

    PubMed

    Branquinho, Raquel; Klein, Günter; Kämpfer, Peter; Peixe, Luísa V

    2015-03-01

    During a study assessing the diversity of the Bacillus pumilus group it became apparent that the type strains of both Bacillus aerophilus and Bacillus stratosphericus were not available from any established culture collection, nor from the authors who originally described them. Therefore, type strains of these species cannot be included in any further scientific studies. It is therefore proposed that the Judicial Commission of the International Committee of Systematics of Prokaryotes place the names Bacillus aerophilus and Bacillus stratosphericus on the list of rejected names, if suitable replacements for the type strains are not found or if neotype strains are not proposed within two years following the publication of this Request for an Opinion. PMID:25479956

  6. Entomopathogenic fungal endophytes

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Fungal endophytes are quite common in nature and some of them have been shown to have adverse effects against insects, nematodes, and plant pathogens. An introduction to fungal endophytes will be presented, followed by a discussion of research aimed at introducing Beauveria bassiana as a fungal endo...

  7. TRICHODERMA ENDOPHYTES OF SAPWOOD

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Trichoderma endophytes occur in sapwood of trunks of Theobroma spp., Cola spp., Fagus sylvatica, Scalesia pedunculata, and in the woody liana Ancistroderma korupensis. Trichoderma is a genus of soil fungi; thus trunks of trees represent a new niche for soil fungi. Trichoderma endophytes are rare in ...

  8. Preliminary Screening of Endophytic Fungi from Medicinal Plants in Malaysia for Antimicrobial and Antitumor Activity

    PubMed Central

    Radu, Son; Kqueen, Cheah Yoke

    2002-01-01

    The screening of antimicrobial activity against Gram-positive bacteria, Gram-negative bacteria, yeast and fungi was carried out on isopropanol extracts prepared from 121 isolates of endophytic fungi isolated from medicinal plants in Malaysia. Sensitivity was found to vary among the microorganisms. Bacillus subtilis, Saccharomyces cerevisiae and Alternaria sp. were susceptible to extracts from three, two and two isolates of endophytic fungi, respectively. None were found effective against Salmonella typhimurium. Sixteen endophytic fungal isolates tested were also found to exhibit antitumor activity in the yeast cell-based assay. PMID:22844221

  9. Surfactin production by strains of Bacillus mojavensis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Bacillus mojavensis, RRC101 is an endophytic bacterium patented for control of fungal diseases in maize and other plants. DNA fingerprint analysis of the rep-PCR fragments of 35 B. mojavensis and 4 B. subtilis strains using the Diversilab genotyping system revealed genotypic distinctive strains alon...

  10. Bacillus DNA in fossil bees: an ancient symbiosis?

    PubMed

    Cano, R J; Borucki, M K; Higby-Schweitzer, M; Poinar, H N; Poinar, G O; Pollard, K J

    1994-06-01

    We report here the isolation of DNA from abdominal tissue of four extinct stingless bees (Proplebeia dominicana) in Dominican amber, PCR amplification of a 546-bp fragment of the 16S rRNA gene from Bacillus spp., and their corresponding nucleotide sequences. These sequences were used in basic local alignment search tool searches of nonredundant nucleic acid data bases, and the highest scores were obtained with 16S rRNA sequences from Bacillus spp. Phylogenetic inference analysis by the maximum-likelihood method revealed close phylogenetic relationships of the four presumed ancient Bacillus sequences with Bacillus pumilus, B. firmus, B. subtilis, and B. circulans. These four extant Bacillus spp. are commonly isolated from abdominal tissue of stingless bees. The close phylogenetic association of the extracted DNA sequences with these bee colonizers suggests that a similar bee-Bacillus association existed in the extinct species P. dominicana. PMID:8031102

  11. Bacillus DNA in fossil bees: an ancient symbiosis?

    PubMed Central

    Cano, R J; Borucki, M K; Higby-Schweitzer, M; Poinar, H N; Poinar, G O; Pollard, K J

    1994-01-01

    We report here the isolation of DNA from abdominal tissue of four extinct stingless bees (Proplebeia dominicana) in Dominican amber, PCR amplification of a 546-bp fragment of the 16S rRNA gene from Bacillus spp., and their corresponding nucleotide sequences. These sequences were used in basic local alignment search tool searches of nonredundant nucleic acid data bases, and the highest scores were obtained with 16S rRNA sequences from Bacillus spp. Phylogenetic inference analysis by the maximum-likelihood method revealed close phylogenetic relationships of the four presumed ancient Bacillus sequences with Bacillus pumilus, B. firmus, B. subtilis, and B. circulans. These four extant Bacillus spp. are commonly isolated from abdominal tissue of stingless bees. The close phylogenetic association of the extracted DNA sequences with these bee colonizers suggests that a similar bee-Bacillus association existed in the extinct species P. dominicana. PMID:8031102

  12. Bacillus species isolated from tungrymbai and bekang, naturally fermented soybean foods of India.

    PubMed

    Chettri, Rajen; Tamang, Jyoti Prakash

    2015-03-16

    Tungrymbai and bekang are naturally fermented soybean foods commonly consumed in Meghalaya and Mizoram states of India. A total of 39 samples of tungrymbai and 43 samples of bekang were collected from different villages and markets of Meghalaya and Mizoram, respectively and were analysed for microbial load. In both tungrymbai and bekang, the average population of Bacillus spp. was 8.2±0.1 log cfu/g. A total of 428 isolates of Bacillus were isolated from tungrymbai (211) and bekang (217) for detailed identification. On the basis of a combination of phenotypic and molecular characterisation using ARDRA, ITS-PCR and RAPD-PCR techniques, species of Bacillus isolated from tungrymbai were identified as Bacillus licheniformis (25.5%), Bacillus pumilus (19.5%) and Bacillus subtilis (55%), and species of Bacillus from bekang were Bacillus brevis (2%), Bacillus circulans (7.5%), Bacillus coagulans (6.5%), B. licheniformis (16.5%), B. pumilus (9.1%), Bacillus sphaericus (4.6%), B. subtilis (51.8%), and Lysinibacillus fusiformis (2%). The most dominant bacterium in both products was B. subtilis. PMID:25574846

  13. Diversity of endophytic bacteria in medicinally important Nepenthes species

    PubMed Central

    Bhore, Subhash J.; Komathi, Vijayan; Kandasamy, Kodi I.

    2013-01-01

    Background: Nepenthes species are used in traditional medicines to treat various health ailments. However, we do not know which types of endophytic bacteria (EB) are associated with Nepenthes spp. Objective: The objective of this study was to isolate and to identify EB associated with Nepenthes spp. Materials and Methods: Surface-sterilized leaf and stem tissues from nine Nepenthes spp. collected from Peninsular Malaysia were used to isolate EB. Isolates were identified using the polymerase chain reaction-amplified 16S ribosomal DNA (rDNA) sequence similarity based method. Results: Cultivable, 96 isolates were analyzed; and the 16S rDNA sequences analysis suggest that diverse bacterial species are associated with Nepenthes spp. Majority (55.2%) of the isolates were from Bacillus genus, and Bacillus cereus was the most dominant (14.6%) among isolates. Conclusion: Nepenthes spp. do harbor a wide array of cultivable endophytic bacteria. PMID:24082746

  14. Microbial Endophytes of Corn

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Increasing recognition is being given to the impact of microbial population dynamics on the general health of biological systems. Building on previous mycological and bacteriological research on agricultural commodities, more advanced technology is expanding our understanding of the “endophytic habi...

  15. Diversity, antimicrobial and antioxidant activities of culturable bacterial endophyte communities in Aloe vera.

    PubMed

    Akinsanya, Mushafau Adewale; Goh, Joo Kheng; Lim, Siew Ping; Ting, Adeline Su Yien

    2015-12-01

    Twenty-nine culturable bacterial endophytes were isolated from surface-sterilized tissues (root, stem and leaf) of Aloe vera and molecularly characterized to 13 genera: Pseudomonas, Bacillus, Enterobacter, Pantoea, Chryseobacterium, Sphingobacterium, Aeromonas, Providencia, Cedecea, Klebsiella, Cronobacter, Macrococcus and Shigella. The dominant genera include Bacillus (20.7%), Pseudomonas (20.7%) and Enterobacter (13.8%). The crude and ethyl acetate fractions of the metabolites of six isolates, species of Pseudomonas, Bacillus, Chryseobacterium and Shigella, have broad spectral antimicrobial activities against pathogenic Pseudomonas aeruginosa, Staphylococcus aureus, Bacillus cereus, Salmonella Typhimurium, Proteus vulgaris, Klebsiella pneumoniae, Escherichia coli, Streptococcus pyogenes and Candida albicans, with inhibition zones ranging from 6.0 ± 0.57 to 16.6 ± 0.57 mm. In addition, 80% of the bacterial endophytes produced 1,1-diphenyl-2-picrylhydrazyl (DPPH) with scavenging properties of over 75% when their crude metabolites were compared with ascorbic acid (92%). In conclusion, this study revealed for the first time the endophytic bacteria communities from A. vera (Pseudomonas hibiscicola, Macrococcus caseolyticus, Enterobacter ludwigii, Bacillus anthracis) that produce bioactive compounds with high DPPH scavenging properties (75-88%) and (Bacillus tequilensis, Pseudomonas entomophila, Chryseobacterium indologenes, Bacillus aerophilus) that produce bioactive compounds with antimicrobial activities against bacterial pathogens. Hence, we suggest further investigation and characterization of their bioactive compounds. PMID:26454221

  16. Establishing fungal entomopathogens as endophytes: towards endophytic biological control

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Beauveria basssiana is a fungal entomopathogen with the ability to colonize plants endophytically. As an endophyte, B. bassiana may play a role in protecting plants from herbivory and disease. This protocol demonstrates two inoculation methods to establish B. bassiana endophytically in the common be...

  17. Reclassification of Bacillus invictae as a later heterotypic synonym of Bacillus altitudinis.

    PubMed

    Liu, Yang; Lai, Qiliang; Du, Juan; Shao, Zongze

    2015-08-01

    The aim of this study was to reclarify the taxonomic status of strain Bacillus invictae Bi.FFUP1T by performing comparative analyses with the other four type strains within the Bacillus pumilus group. The digital DNA-DNA hybridization (dDDH) and average nucleotide identity (ANI) values between strains B. invictae Bi.FFUP1T (?=?DSMZ 26896T?=?MCCC 1A07089T), B. altitudinis 41KF2bT (?=?DSMZ 21631T?=?MCCC 1A06452T), B. safensis FO-36bT (?=?DSMZ 19292T?=?MCCC 1A6451T), B. pumilus ATCC 7061T (?=?DSMZ 27T?=?MCCC 1A06453T) and B. xiamenensis HYC-10T (?=?MCCC 1A00008T) were, respectively, 82.90??% and 98.10??%, which are greater than the thresholds for bacterial species delineation, suggesting that they should belong to the same species, while the dDDH and ANI values between strain B. invictae DSMZ 26896T and the other three type strains within the B. pumilus group were below the respective thresholds of 70??% and 95??%. Meanwhile, B. invictae DSMZ 26896T and B. altitudinis 41KF2bT shared 98.7??% gyrB gene sequence similarity based on resequencing, whereas strain B. invictae DSMZ 26896T shared low similarities (?Bacillus invictae Branquinho et al. 2014 should be regarded as a later heterotypic synonym of Bacillus altitudinisShivaji et al. 2006. PMID:25977285

  18. Cultivable endophytic bacteria from leaf bases of Agave tequilana and their role as plant growth promoters.

    PubMed

    Martínez-Rodríguez, Julia del C; De la Mora-Amutio, Marcela; Plascencia-Correa, Luis A; Audelo-Regalado, Esmeralda; Guardado, Francisco R; Hernández-Sánchez, Elías; Peña-Ramírez, Yuri J; Escalante, Adelfo; Beltrán-García, Miguel J; Ogura, Tetsuya

    2014-01-01

    Agave tequilana Weber var. 'Azul' is grown for the production of tequila, inulin and syrup. Diverse bacteria inhabit plant tissues and play a crucial role for plant health and growth. In this study culturable endophytic bacteria were extracted from leaf bases of 100 healthy Agave tequilana plants. In plant tissue bacteria occurred at mean population densities of 3 million CFU/g of fresh plant tissue. Three hundred endophytic strains were isolated and 16s rDNA sequences grouped the bacteria into eight different taxa that shared high homology with other known sequences. Bacterial endophytes were identified as Acinectobacter sp., A. baumanii, A. bereziniae, Cronobacter sakazakii, Enterobacter hormaechei, Bacillus sp. Klebsiella oxytoca, Pseudomonas sp., Enterococcus casseliflavus, Leuconostoc mesenteroides subsp. mesenteroides and Gluconobacter oxydans. Isolates were confirmed to be plant growth promoting bacteria (PGPB) by their capacities for nitrogen fixation, auxin production, phosphate solubilization, or antagonism against Fusarium oxysporum AC132. E. casseliflavus JM47 and K. oxytoca JM26 secreted the highest concentrations of IAA. The endophyte Acinectobacter sp. JM58 exhibited the maximum values for nitrogen fixation and phosphate solubilization index (PSI). Inhibition of fungi was found in Pseudomonas sp. JM9p and K. oxytoca JM26. Bacterial endophytes show promise for use as bio-inoculants for agave cultivation. Use of endophytes to enhance cultivation of agave may be particularly important for plants produced by micropropagation techniques, where native endophytes may have been lost. PMID:25763038

  19. Cultivable endophytic bacteria from leaf bases of Agave tequilana and their role as plant growth promoters

    PubMed Central

    Martínez-Rodríguez, Julia del C.; la Mora-Amutio, Marcela De; Plascencia-Correa, Luis A.; Audelo-Regalado, Esmeralda; Guardado, Francisco R.; Hernández-Sánchez, Elías; Peña-Ramírez, Yuri J.; Escalante, Adelfo; Beltrán-García, Miguel J.; Ogura, Tetsuya

    2014-01-01

    Agave tequilana Weber var. ‘Azul’ is grown for the production of tequila, inulin and syrup. Diverse bacteria inhabit plant tissues and play a crucial role for plant health and growth. In this study culturable endophytic bacteria were extracted from leaf bases of 100 healthy Agave tequilana plants. In plant tissue bacteria occurred at mean population densities of 3 million CFU/g of fresh plant tissue. Three hundred endophytic strains were isolated and 16s rDNA sequences grouped the bacteria into eight different taxa that shared high homology with other known sequences. Bacterial endophytes were identified as Acinectobacter sp., A. baumanii, A. bereziniae, Cronobacter sakazakii, Enterobacter hormaechei, Bacillus sp. Klebsiella oxytoca, Pseudomonas sp., Enterococcus casseliflavus, Leuconostoc mesenteroides subsp. mesenteroides and Gluconobacter oxydans. Isolates were confirmed to be plant growth promoting bacteria (PGPB) by their capacities for nitrogen fixation, auxin production, phosphate solubilization, or antagonism against Fusarium oxysporum AC132. E. casseliflavus JM47 and K. oxytoca JM26 secreted the highest concentrations of IAA. The endophyte Acinectobacter sp. JM58 exhibited the maximum values for nitrogen fixation and phosphate solubilization index (PSI). Inhibition of fungi was found in Pseudomonas sp. JM9p and K. oxytoca JM26. Bacterial endophytes show promise for use as bio-inoculants for agave cultivation. Use of endophytes to enhance cultivation of agave may be particularly important for plants produced by micropropagation techniques, where native endophytes may have been lost. PMID:25763038

  20. Microbial endophytes: future challenges

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Endophytes are represented by a diverse group of prokaryotic (bacteria or cyanobacteria) or eukaryotic (fungi or parasitic vascular plants) organisms that form life-long associations within tissues of plants. Ecologically, these associations are usually viewed as advantageous although in some insta...

  1. Isolation and enzyme bioprospection of endophytic bacteria associated with plants of Brazilian mangrove ecosystem.

    PubMed

    Castro, Renata A; Quecine, Maria Carolina; Lacava, Paulo T; Batista, Bruna D; Luvizotto, Danice M; Marcon, Joelma; Ferreira, Anderson; Melo, Itamar S; Azevedo, João L

    2014-01-01

    The mangrove ecosystem is a coastal tropical biome located in the transition zone between land and sea that is characterized by periodic flooding, which confers unique and specific environmental conditions on this biome. In these ecosystems, the vegetation is dominated by a particular group of plant species that provide a unique environment harboring diverse groups of microorganisms, including the endophytic microorganisms that are the focus of this study. Because of their intimate association with plants, endophytic microorganisms could be explored for biotechnologically significant products, such as enzymes, proteins, antibiotics and others. Here, we isolated endophytic microorganisms from two mangrove species, Rhizophora mangle and Avicennia nitida, that are found in streams in two mangrove systems in Bertioga and Cananéia, Brazil. Bacillus was the most frequently isolated genus, comprising 42% of the species isolated from Cananéia and 28% of the species from Bertioga. However, other common endophytic genera such as Pantoea, Curtobacterium and Enterobacter were also found. After identifying the isolates, the bacterial communities were evaluated for enzyme production. Protease activity was observed in 75% of the isolates, while endoglucanase activity occurred in 62% of the isolates. Bacillus showed the highest activity rates for amylase and esterase and endoglucanase. To our knowledge, this is the first reported diversity analysis performed on endophytic bacteria obtained from the branches of mangrove trees and the first overview of the specific enzymes produced by different bacterial genera. This work contributes to our knowledge of the microorganisms and enzymes present in mangrove ecosystems. PMID:25110630

  2. [The identification of Bacillus cereus, Bacillus lichenformis and Bacillus subtilis strains using the coagglutination reaction].

    PubMed

    Hellmann, E; Stanzel, C; Tonkaboni, T F

    1994-09-01

    Polyclonal rabbit antisera against 4 B. cereus strains were selected from a total of 9 B. cereus antisera and pooled. This serum agglutinated all available B. cereus strains (n = 63) at a titre > or = 1:64 when tested by a slide co-agglutination reaction. One hundred and thirty-six Bacillus strains belonging to 17 other species reacted with higher serum concentrations only (titres mainly < or = 1:16). A pooled antiserum comprised of two B. licheniformis antisera and two B. subtilis antisera agglutinated all 43 B. licheniformis strains and all 38 B. subtilis strains at a titre > or = 1:128, but one strain each of the species B. pumilus, B. macerans and B. fastidiosus was also agglutinated at a titre 1:128. PMID:7980381

  3. Endophytic bacteria from banana cultivars and their antifungal activity.

    PubMed

    Souza, A; Cruz, J C; Sousa, N R; Procópio, A R L; Silva, G F

    2014-01-01

    Endophytic microorganisms consist of fungi, bacteria, and actinomycetes that play important roles in the process of plant adaptation to the environment. Currently, the natural associations between microorganisms and plant species are being explored for a large number of biotechnological applications. In this study, 122 endophytic bacteria were isolated from 5 cultivars of Musa spp from the state of Amazonas (Brazil). Four strains were selected because they exhibited antagonistic activities against Fusarium oxysporum f. sp cubense and Colletotrichum guaranicola, with inhibitions ranging from 19 to 30% and 27 to 35%, respectively. Phylogenetic analysis of the 16S rDNA regions of these bacteria with antifungal activity showed that they are phylogenetically related to 3 different species of Bacillus - B. amyloliquefaciens, B. subtilis subsp subtilis, and B. thuringiensis. PMID:25366756

  4. Grass fungal endophytes and uses thereof

    SciTech Connect

    Craven, Kelly

    2015-03-10

    The invention provides isolated fungal endophytes and synthetic combinations thereof with host grass plants. Methods for inoculating grass plant with the endophytes, for propagating the grass-endophyte combinations, and for producing feeds and biofuels from grass-endophyte combinations are also provided.

  5. Antiproliferative, antifungal, and antibacterial activities of endophytic alternaria species from cupressaceae.

    PubMed

    Soltani, Jalal; Hosseyni Moghaddam, Mahdieh S

    2014-09-01

    Recent research has shown the bioprospecting of endophytic fungi from Cupressaceae. Here, we further uncover that the healthy cypress plants such as Cupressus arizonica, Cupressus sempervirens var. cereiformis, and Thuja orientalis host highly bioactive endophytic Alternaria fungal species. Indeed, endophytic Alternaria alternata, Alternaria pellucida, and Alternaria tangelonis were recovered from healthy Cupressaceous trees. Biodiversity and bioactivity of recovered endophytic Alternaria species were a matter of biogeography and host identity. We further extracted such Alternaria's metabolites and highlighted their significant antiproliferative, growth inhibitory, and antibacterial activities against the model target fungus Pyricularia oryzae and the model pathogenic bacteria Bacillus sp., Erwinia amylovora, and Pseudomonas syringae. In vitro assays also indicated that endophytic Alternaria species significantly inhibited the growth of cypress fungal phytopathogens Diplodia seriata, Phaeobotryon cupressi, and Spencermartinsia viticola. In conclusion, since the recovered Alternaria species were originally reported as pathogenic and allergenic fungi, our findings suggest a possible ecological niche for them inside the foliar tissues of Cupressaceous trees. Moreover, in this study, the significant bioactivities of endophytic Alternaria species in association with Cupressaceae plant family are reported. PMID:24801337

  6. Bacillus safensis sp. nov., isolated from spacecraft and assembly-facility surfaces.

    PubMed

    Satomi, Masataka; La Duc, Myron T; Venkateswaran, Kasthuri

    2006-08-01

    Thirteen strains of a novel spore-forming, Gram-positive, mesophilic heterotrophic bacterium were isolated from spacecraft surfaces (Mars Odyssey Orbiter) and assembly-facility surfaces at the Jet Propulsion Laboratory in California and the Kennedy Space Center in Florida. Phylogenetic analysis of 16S rRNA gene sequences has placed these novel isolates within the genus Bacillus, the greatest sequence similarity (99.9 %) being found with Bacillus pumilus. However, these isolates share a mere 91.2 % gyrB sequence similarity with Bacillus pumilus, rendering their 16S rRNA gene-derived relatedness suspect. Furthermore, DNA-DNA hybridization showed only 54-66 % DNA relatedness between the novel isolates and strains of B. pumilus. rep-PCR fingerprinting and previously reported matrix-assisted laser desorption/ionization time-of-flight mass spectrometry protein profiling clearly distinguished these isolates from B. pumilus. Phenotypic analyses also showed some differentiation between the two genotypic groups, although the fatty acid compositions were almost identical. The polyphasic taxonomic studies revealed distinct clustering of the tested strains into two distinct species. On the basis of phenotypic characteristics and the results of phylogenetic analyses of 16S rRNA and gyrB gene sequences, repetitive element primer-PCR fingerprinting and DNA-DNA hybridization, the 13 isolates represent a novel species of the genus Bacillus, for which the name Bacillus safensis sp. nov. is proposed. The type strain is FO-36b(T) (=ATCC BAA-1126(T)=NBRC 100820(T)). PMID:16902000

  7. Novel extremely acidic lipases produced from Bacillus species using oil substrates.

    PubMed

    Saranya, P; Kumari, H Sukanya; Jothieswari, M; Rao, B Prasad; Sekaran, G

    2014-01-01

    The extremely acidophilic microorganisms Bacillus pumilus and Bacillus subtilis were isolated from soil collected from the commercial edible oil and fish oil extraction industry. Optimization of conditions for acidic lipase production from B. pumilus and B. subtilis using palm oil and fish oil, respectively, was carried out using response surface methodology. The extremely acidic lipases, thermo-tolerant acidic lipase (TAL) and acidic lipase (AL), were produced by B. pumilus and B. subtilis, respectively. The optimum conditions for B. pumilus obtaining the maximum activity (1,100 U/mL) of TAL were fermentation time, 96 h; pH, 1; temperature, 50 °C; concentration of palm oil, 50 g/L. After purification, a 7.1-fold purity of lipase with specific activity of 5,173 U/mg protein was obtained. The molecular weight of the TAL was 55 kDa. The AL from B. subtilis activity was 214 U/mL at a fermentation time of 72 h; pH, 1; temperature, 35 °C; concentration of fish oil, 30 g/L; maltose concentration, 10 g/L. After purification, an 11.4-fold purity of lipase with specific activity of 2,189 U/mg protein was obtained. The molecular weight of the extremely acidic lipase was 22 kDa. The functional groups of lipases were determined by Fourier transform-infrared (FT-IR) spectroscopy. PMID:24185617

  8. Action of antimicrobial substances produced by different oil reservoir Bacillus strains against biofilm formation.

    PubMed

    Korenblum, E; Sebastián, G V; Paiva, M M; Coutinho, C M L M; Magalhães, F C M; Peyton, B M; Seldin, L

    2008-05-01

    Microbial colonization of petroleum industry systems takes place through the formation of biofilms, and can result in biodeterioration of the metal surfaces. In a previous study, two oil reservoir Bacillus strains (Bacillus licheniformis T6-5 and Bacillus firmus H(2)O-1) were shown to produce antimicrobial substances (AMS) active against different Bacillus strains and a consortium of sulfate-reducing bacteria (SRB) on solid medium. However, neither their ability to form biofilms nor the effect of the AMS on biofilm formation was adequately addressed. Therefore, here, we report that three Bacillus strains (Bacillus pumilus LF4 -- used as an indicator strain, B. licheniformis T6-5, and B. firmus H(2)O-1), and an oil reservoir SRB consortium (T6lab) were grown as biofilms on glass surfaces. The AMS produced by strains T6-5 and H(2)O-1 prevented the formation of B. pumilus LF4 biofilm and also eliminated pre-established LF4 biofilm. In addition, the presence of AMS produced by H(2)O-1 reduced the viability and attachment of the SRB consortium biofilm by an order of magnitude. Our results suggest that the AMS produced by Bacillus strains T6-5 and H(2)O-1 may have a potential for pipeline-cleaning technologies to inhibit biofilm formation and consequently reduce biocorrosion. PMID:18330565

  9. Genetic discrimination of foodborne pathogenic and spoilage Bacillus spp. based on three housekeeping genes.

    PubMed

    Caamaño-Antelo, S; Fernández-No, I C; Böhme, K; Ezzat-Alnakip, M; Quintela-Baluja, M; Barros-Velázquez, J; Calo-Mata, P

    2015-04-01

    Bacillus genus includes foodborne pathogenic and spoilage-associated species, such as Bacillus cereus, Bacillus licheniformis, Bacillus subtilis and Bacillus pumilus. Bacillus is also a heterogeneous genus that includes closely related species that are difficult to discriminate among, especially when well-conserved genes such as 16S rRNA and 23S rRNA are considered. The main goal of the present work was to study the usefulness of three housekeeping genes, the TU elongation factor (tuf), the DNA gyrase ? subunit (gyrB) and the RNA polymerase ? subunit (rpoB) genes, for use in differentiating among the most important foodborne Bacillus spp. sequences from 20 foodborne isolated Bacillus strains, and sequences belonging to different Bacillus spp. retrieved from the GenBank were analysed. In general terms, gyrB, rpoB and tuf gene regions for the strains considered in this study exhibited interspecific similarities of 57.8%, 67.23% and 77.66% respectively. Novel tufGPF and tufGPR universal primers targeted to the tuf gene were designed and proved to be useful for the amplification of all Bacillus spp considered. In conclusion, the tuf gene can be considered to be a good target for the differential characterisation of foodborne Bacillus species, especially for differentiating B. subtilis and B. cereus from other closely related species. PMID:25475298

  10. Rapid inactivation of seven Bacillus spp. under simulated Mars UV irradiation

    NASA Astrophysics Data System (ADS)

    Schuerger, Andrew C.; Richards, Jeff T.; Newcombe, David A.; Venkateswaran, Kasthuri

    2006-03-01

    Seven Bacillus spp. were exposed to simulations of Mars-normal UV fluence rates in order to study the effects of UV irradiation on microbial survival. A UV illumination system was calibrated to deliver 9.78 W m -2 (35.2 kJ m -2 h -1) of UVC + UVB irradiation (200-320 nm) to microbial samples, thus creating a clear-sky simulation (0.5 optical depth) of equatorial Mars. The Bacillus spp. studied were: B. licheniformis KL-196, B. megaterium KL-197, B. nealsonii FO-092, B. pumilus FO-36B, B. pumilus SAFR-032, B. subtilis 42HS1, and B. subtilis HA101. The bacteria were prepared as thin monolayers of endospores on aluminum coupons in order to simulate contaminated spacecraft surfaces. Bacterial monolayers were exposed to Mars UV irradiation for time-steps of 0, 0.25, 0.5, 1, 5, 15, 30, 60, 120, or 180 min. The surviving endospores were then assayed with a Most Probable Numbers (MPN) procedure and with a culture-based assay that utilized a bacillus spore germination medium. Results indicated that B. pumilus SAFR-032 was the most resistant, and B. subtilis 42HS-1 and B. megaterium were the most sensitive of the seven strains exposed to martian UV fluence rates. Bacillus subtilis 42HS1 and B. megaterium were inactivated after 30 min exposure to Mars UV, while B. pumilus SAFR-032 required 180 min for full inactivation in both assays. Spores of B. pumilus SAFR-032 exhibited significantly different inactivation kinetics suggesting that this wild type isolate also was more resistant than the standard dosimetric strain, B. subtilis HA101. Although the various Bacillus spp. exhibited diverse levels of UV resistance, none were immune to UV irradiation, and, thus, all species would be expected to be inactivated on Sun-exposed spacecraft surfaces within a few tens-of-minutes to a few hours on sol 1 under clear-sky conditions on equatorial Mars. The inactivation kinetics of all seven Bacillus spp. support the conclusion that significant levels of bioload reductions are possible on Sun-exposed spacecraft surfaces in very short time periods under clear-sky conditions on Mars. However, the presence of UV resistant microbes on spacecraft surfaces rapidly covered in dust during landing operations, and non-Sun-exposed surfaces of spacecraft remain concerns that must continue to be addressed through adequate spacecraft sanitizing procedures prior to launch.

  11. The contribution of endophytic bacteria to Albizia lebbeck-mediated phytoremediation of tannery effluent contaminated soil.

    PubMed

    Manikandan, Muthu; Kannan, Vijayaraghavan; Mendoza, Ordetta Hannah; Kanimozhi, Mahalingam; Chun, Sechul; Pašić, Lejla

    2016-01-01

    Toxicity of chromium often impairs the remediation capacity of plants used in phytoremediation of polluted soils. In this study, we have identified Albizia lebbeck as a prospective chromium hyperaccumulator and examined cultivable diversity of endophytes present in chromium-treated and control saplings. High numbers (22-100%) of endophytic bacteria, isolated from root, stem, and leaf tissues, could tolerate elevated (1-3 mM) concentrations of K2CrO7. 16S rRNA gene sequence-based phylogenetic analysis showed that the 118 isolates obtained comprised of 17 operational taxonomic units affiliated with the proteobacterial genera Rhizobium (18%), Marinomonas (1%), Pseudomonas (16%), and Xanthomonas (7%) but also with members of Firmicutes genera, such as Bacillus (35%) and Salinococcus (3%). The novel isolates belonging to Salinococcus and Bacillus could tolerate high K2CrO7 concentrations (3 mM) and also showed elevated activity of chromate reductase. In addition, majority (%) of the endophytic isolates also showed production of indole-3-acetic acid. Taken together, our results indicate that the innate endophytic bacterial community assists plants in reducing heavy metal toxicity. PMID:26147743

  12. Establishing Fungal Entomopathogens as Endophytes: Towards Endophytic Biological Control

    PubMed Central

    Parsa, Soroush; Ortiz, Viviana; Vega, Fernando E.

    2013-01-01

    Beauveria bassiana is a fungal entomopathogen with the ability to colonize plants endophytically. As an endophyte, B. bassiana may play a role in protecting plants from herbivory and disease. This protocol demonstrates two inoculation methods to establish B. bassiana endophytically in the common bean (Phaseolus vulgaris), in preparation for subsequent evaluations of endophytic biological control. Plants are grown from surface-sterilized seeds for two weeks before receiving a B. bassiana treatment of 108 conidia/ml (or water) applied either as a foliar spray or a soil drench. Two weeks later, the plants are harvested and their leaves, stems and roots are sampled to evaluate endophytic fungal colonization. For this, samples are individually surface sterilized, cut into multiple sections, and incubated in potato dextrose agar media for 20 days. The media is inspected every 2-3 days to observe fungal growth associated with plant sections and record the occurrence of B. bassiana to estimate the extent of its endophytic colonization. Analyses of inoculation success compare the occurrence of B. bassiana within a given plant part (i.e. leaves, stems or roots) across treatments and controls. In addition to the inoculation method, the specific outcome of the experiment may depend on the target crop species or variety, the fungal entomopathogen species strain or isolate used, and the plant's growing conditions. PMID:23603853

  13. Bacillus invictae sp. nov., isolated from a health product.

    PubMed

    Branquinho, Raquel; Sousa, Clara; Osório, Hugo; Meirinhos-Soares, Luís; Lopes, João; Carriço, João A; Busse, Hans-Jürgen; Abdulmawjood, Amir; Klein, Günter; Kämpfer, Peter; Pintado, Manuela E; Peixe, Luísa V

    2014-11-01

    A Gram-positive, rod-shaped, endospore-forming Bacillus isolate, Bi.(FFUP1) (T), recovered in Portugal from a health product was subjected to a polyphasic study and compared with the type strains of Bacillus pumilus, Bacillus safensis, Bacillus altitudinis and Bacillus xiamenensis, the phenotypically and genotypically most closely related species. Acid production from cellobiose, D-glucose and D-mannose and absence of acid production from D-arabinose, erythritol, inositol, maltose, mannitol, raffinose, rhamnose, sorbitol, starch and L-tryptophan discriminated this new isolate from the type strains of the most closely related species. Additionally, a significant different protein and carbohydrate signature was evidenced by spectroscopic techniques, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and Fourier transform IR spectroscopy with attenuated total reflectance. Using a chemometric approach, the score plot generated by principal component analysis clearly delineated the isolate as a separate cluster. The quinone system for strain Bi.(FFUP1) (T) comprised predominantly menaquinone MK-7 and major polar lipids were diphosphatidylglycerol, an unidentified phospholipid and an unidentified glycolipid. Strain Bi.(FFUP1) (T) showed ? 99% 16S rRNA gene sequence similarity to B. safensis FO-036b(T), B. pumilus (7061(T) and SAFR-032), B. altitudinis 41KF2b(T) and B. xiamenensis HYC-10(T). Differences in strain Bi.FFUP1 (T) gyrB and rpoB sequences in comparison with the most closely related species and DNA-DNA hybridization experiments with Bi.FFUP1 (T) and B. pumilus ATCC 7061(T), B. safensis FO-036b(T), B. altitudinis 41KF2b(T) and B. xiamenensis HYC-10(T) gave relatedness values of 39.6% (reciprocal 38.0%), 49.9% (reciprocal 42.9%), 61.9% (reciprocal 52.2%) and 61.7% (reciprocal 49.2%), respectively, supported the delineation of strain Bi.(FFUP1) (T) as a representative of a novel species of the genus Bacillus, for which the name Bacillus invictae sp. nov. is proposed, with strain Bi.(FFUP1) (T) (?=DSM 26896(T)?=CCUG 64113(T)) as the type strain. PMID:25171924

  14. Response of endophytic bacterial communities in banana tissue culture plantlets to Fusarium wilt pathogen infection.

    PubMed

    Lian, Jie; Wang, Zifeng; Zhou, Shining

    2008-04-01

    Endophytic bacteria reside within plant hosts without having pathogenic effects, and various endophytes have been found to functionally benefit plant disease suppressive ability. In this study, the influence of banana plant stress on the endophytic bacterial communities, which was achieved by infection with the wilt pathogen Fusarium oxysporum f. sp. cubense, was examined by cultivation-independent denaturing gradient gel electrophoresis analysis of 16S ribosomal DNA directly amplified from plant tissue DNA. Community analysis clearly demonstrated increased bacterial diversity in pathogen-infected plantlets compared to that in control plantlets. By sequencing, bands most similar to species of Bacillus and Pseudomonas showed high density in the pathogen-treated pattern. In vitro screening of the isolates for antagonistic activity against Fusarium wilt pathogen acquired three strains of endophytic bacteria which were found to match those species that obviously increased in the pathogen infection process; moreover, the most inhibitive strain could also interiorly colonize plantlets and perform antagonism. The evidence obtained from this work showed that antagonistic endophytic bacteria could be induced by the appearance of a host fungal pathogen and further be an ideal biological control agent to use in banana Fusarium wilt disease protection. PMID:18497482

  15. Endophytic bacterial diversity in banana 'Prata Anã' (Musa spp.) roots.

    PubMed

    Souza, Suzane A; Xavier, Adelica A; Costa, Márcia R; Cardoso, Acleide M S; Pereira, Marlon C T; Nietsche, Silvia

    2013-07-01

    The genetic diversity of endophytic bacteria in banana 'Prata Anã' roots was characterized. Two hundred and one endophytic bacteria were isolated, 151 of which were classified as Gram-positive and 50 as Gram-negative. No hypersensitivity response was observed in any of the isolates. The rep-PCR technique generated different molecular profiles for each primer set (REP, ERIC and BOX). Fifty readable loci were obtained and all of the fragments were polymorphic. Amplified ribosomal DNA restriction analysis (ARDRA) of the isolates based on cleavage with four restriction enzymes yielded 45 polymorphic bands and no monomorphic bands. PCR amplified the nifH gene in 24 isolates. 16S rDNA sequencing of the 201 bacterial isolates yielded 102 high-quality sequences. Sequence analyses revealed that the isolates were distributed among ten bacterial genera (Agrobacterium, Aneurinibacillus, Bacillus, Enterobacter, Klebsiella, Lysinibacillus, Micrococcus, Paenibacillus, Rhizobium and Sporolactobacillus) and included 15 species. The greatest number of isolates belonged to the genus Bacillus. The bacteria identified in this study may be involved in promoting growth, phosphate solubilization, biological control and nitrogen fixation in bananas. PMID:23885208

  16. Endophytic bacterial diversity in banana ‘Prata An㒠(Musa spp.) roots

    PubMed Central

    Souza, Suzane A.; Xavier, Adelica A.; Costa, Márcia R.; Cardoso, Acleide M.S.; Pereira, Marlon C.T.; Nietsche, Silvia

    2013-01-01

    The genetic diversity of endophytic bacteria in banana ‘Prata An㒠roots was characterized. Two hundred and one endophytic bacteria were isolated, 151 of which were classified as Gram-positive and 50 as Gram-negative. No hypersensitivity response was observed in any of the isolates. The rep-PCR technique generated different molecular profiles for each primer set (REP, ERIC and BOX). Fifty readable loci were obtained and all of the fragments were polymorphic. Amplified ribosomal DNA restriction analysis (ARDRA) of the isolates based on cleavage with four restriction enzymes yielded 45 polymorphic bands and no monomorphic bands. PCR amplified the nifH gene in 24 isolates. 16S rDNA sequencing of the 201 bacterial isolates yielded 102 high-quality sequences. Sequence analyses revealed that the isolates were distributed among ten bacterial genera (Agrobacterium, Aneurinibacillus, Bacillus, Enterobacter, Klebsiella, Lysinibacillus, Micrococcus, Paenibacillus, Rhizobium and Sporolactobacillus) and included 15 species. The greatest number of isolates belonged to the genus Bacillus. The bacteria identified in this study may be involved in promoting growth, phosphate solubilization, biological control and nitrogen fixation in bananas. PMID:23885208

  17. Surfactin A Production and Isoform Characterizations in Strains of Bacillus mojavensis for Potential Control of Fusarium verticillioides and Fumonisin in Maize

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Bacillus mojavensis, a species recently distinguished as a cryptic species within Bacillus subtilis, was discovered in maize kernels and later determined to possess endophytic characteristics. The bacterium was also determined to have biocontrol potential due to its strong antagonism to the fungus...

  18. Habitat filters in fungal endophyte community assembly

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Fungal endophytes can influence host health, and more broadly, can instigate trophic cascades with effects scaling to the ecosystem level. Despite this, biotic mechanisms of endophyte community assembly are largely unknown. We used maize to investigate three potential habitat filters in endophyte co...

  19. Metabolic potential and community structure of endophytic and rhizosphere bacteria associated with the roots of the halophyte Aster tripolium L.

    PubMed

    Szymańska, Sonia; Płociniczak, Tomasz; Piotrowska-Seget, Zofia; Złoch, Michał; Ruppel, Silke; Hrynkiewicz, Katarzyna

    2016-01-01

    The submitted work assumes that the abundance and diversity of endophytic and rhizosphere microorganisms co-existing with the halophytic plant Aster tripolium L. growing in a salty meadow in the vicinity of a soda factory (central Poland) represent unique populations of cultivable bacterial strains. Endophytic and rhizosphere bacteria were (i) isolated and identified based on 16S rDNA sequences; (ii) screened for nifH and acdS genes; and (iii) analyzed based on selected metabolic properties. Moreover, total microbial biomass and community structures of the roots (endophytes), rhizosphere and soil were evaluated using a cultivation-independent technique (PLFA) to characterize plant-microbial interactions under natural salt conditions. The identification of the isolated strains showed domination by Gram-positive bacteria (mostly Bacillus spp.) both in the rhizosphere (90.9%) and roots (72.7%) of A. tripolium. Rhizosphere bacterial strains exhibited broader metabolic capacities, while endophytes exhibited higher specificities for metabolic activity. The PLFA analysis showed that the total bacterial biomass decreased in the following order (rhizosphereendophytes) and confirmed the dominance of Gram-positive endophytic bacteria in the roots of the halophyte. The described strain collection provides a valuable basis for a subsequent applications of bacteria in improvement of site adaptation of plants in saline soils. PMID:26686615

  20. Plant Growth Promoting of Endophytic Sporosarcina aquimarina SjAM16103 Isolated from the Pneumatophores of Avicennia marina L.

    PubMed Central

    Janarthine, S. Rylo Sona; Eganathan, P.

    2012-01-01

    Endophytic Sporosarcina aquimarina SjAM16103 was isolated from the inner tissues of pneumatophores of mangrove plant Avicennia marina along with Bacillus sp. and Enterobacter sp. Endophytic S. aquimarina SjAM16103 was Gram variable, and motile bacterium measured 0.6–0.9??m wide by 1.7–2.0??m long and light orange-brown coloured in 3-day cultures on tryptone broth at 26°C. Nucleotide sequence of this strain has been deposited in the GenBank under accession number GU930359. This endophytic bacterium produced 2.37??Mol/mL of indole acetic acid and siderophore as it metabolites. This strain could solubilize phosphate molecules and fixes atmospheric nitrogen. Endophytic S. aquimarina SjAM16103 was inoculated into four different plants under in vitro method to analyse its growth-promoting activity and role inside the host plants. The growth of endophytic S. aquimarina SjAM16103 inoculated explants were highly significant than the uninoculated control explants. Root hairs and early root development were observed in the endophytic S. aquimarina SjAM16103 inoculated explants. PMID:22811715

  1. Bacillus coagulans

    MedlinePLUS

    ... vaccines to improve their effectiveness. Bacillus coagulans produces lactic acid and, as a result, is often misclassified as lactic acid bacteria such as lactobacillus. In fact, some commercial ...

  2. Molecular phylogenetic diversity of Bacillus community and its temporal-spatial distribution during the swine manure of composting.

    PubMed

    Yi, Jing; Wu, Hua-Yu; Wu, Jian; Deng, Chang-Yan; Zheng, Rong; Chao, Zhe

    2012-01-01

    In order to obtain the diversity and temporal-spatial distribution of Bacillus community during the swine manure composting, we utilized traditional culture methods and the modern molecular biology techniques of polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and -denaturing gradient gel electrophoresis (PCR-DGGE). Bacillus species were firstly isolated from the composting. Based on temperature changes, the temporal-spatial characteristics of total culturable Bacillus were remarkable that the number of the culturable Bacillus detected at the high-temperature stage was the highest in each layer of the pile and that detected in the middle layer was the lowest at each stage of composting respectively. The diversity of cultivated Bacillus species isolated from different composting stages was low. A total of 540 isolates were classified by the RFLP method and partial 16S rDNA sequences. They affiliated to eight species including Bacillus subtilis, Bacillus cereus, Bacillus thuringiensis, Bacillus anthracis, Bacillus megaterium, Bacillus licheniformis, Bacillus pumilus, and Bacillus circulans. The predominant species was B. subtilis, and the diversity of culturable Bacillus isolated in the middle-level samples at temperature rising and cooling stages was the highest. The DGGE profile and clone library analysis revealed that the temporal-spatial distribution of Bacillus community was not obvious, species belonging to the Bacillus were dominant (67%) with unculturable bacteria and B. cereus was the second major culturable Bacillus species. This study indicated that a combination of culture and culture-independent approaches could be very useful for monitoring the diversity and temporal-spatial distribution of Bacillus community during the composting process. PMID:21701982

  3. 77 FR 19109 - Bacillus Pumilus Strain GHA 180; Exemption From the Requirement of a Tolerance

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-03-30

    ...- 5805. II. Background and Statutory Findings In the Federal Register of September 30, 2010 (75 FR 60452....: ASM Press; 9th edition, 2007. 4. Doyle, M.P., L.R. Beuchat and T.J. Montville. 1997. Food Microbiology... (58 FR 51735, October 4, 1993). Because this final rule has been exempted from review under...

  4. 78 FR 35147 - Bacillus pumilus Strain BU F-33; Exemption From the Requirement of a Tolerance

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-06-12

    ... or on all food commodities when applied to elicit induced systemic resistance in plants and used in... FR 59578) (FRL- 9364-6), the EPA issued a notice pursuant to FFDCA section 408(d)(3), 21 U.S.C. 346a... strain BU F-33 in or on all food commodities when applied to elicit induced systemic resistance in...

  5. Distribution of Endophytic Bacteria in Alopecurus aequalis Sobol and Oxalis corniculata L. from Soils Contaminated by Polycyclic Aromatic Hydrocarbons

    PubMed Central

    Peng, Anping; Liu, Juan; Gao, Yanzheng; Chen, Zeyou

    2013-01-01

    The distributions of endophytic bacteria in Alopecurus aequalis Sobol and Oxalis corniculata L. grown in soils contaminated with different levels of polycyclic aromatic hydrocarbons (PAHs) were investigated with polymerase chain reaction followed by denaturing gradient gel electrophoresis technology (PCR-DGGE) and cultivation methods. Twelve types of PAHs, at concentrations varying from 0.16 to 180 mg·kg−1, were observed in the roots and shoots of the two plants. The total PAH concentrations in Alopecurus aequalis Sobol obtained from three different PAH-contaminated stations were 184, 197, and 304 mg·kg−1, and the total PAH concentrations in Oxalis corniculata L. were 251, 346, and 600 mg·kg−1, respectively. The PCR-DGGE results showed that the endophytic bacterial communities in the roots and shoots of the two plants were quite different, although most bacteria belonged to Firmicutes, Proteobacteria, Actinobacteria and Bacteroidetes. A total of 68 endophytic bacterial strains were isolated from different tissues of the two plants and classified into three phyla: Firmicutes, Proteobacteria and Bacteroidetes. In both plants, Bacillus spp. and Pseudomonas spp. were the dominant cultivable populations. With an increase in the PAH pollution level, the diversity and distribution of endophytic bacteria in the two plants changed correspondingly, and the number of cultivable endophytic bacterial strains decreased rapidly. Testing of the isolated endophytic bacteria for tolerance to each type of PAH showed that most isolates could grow well on Luria-Bertani media in the presence of different PAHs, and some isolates were able to grow rapidly on a mineral salt medium with a single PAH as the sole carbon and energy source, indicating that these strains may have the potential to degrade PAHs in plants. This research provides the first insight into the characteristics of endophytic bacterial populations under different PAH pollution levels and provides a species resource for the isolation of PAH-degrading endophytic bacteria. PMID:24358247

  6. Plant growth-promoting bacterial endophytes.

    PubMed

    Santoyo, Gustavo; Moreno-Hagelsieb, Gabriel; Del Carmen Orozco-Mosqueda, Ma; Glick, Bernard R

    2016-02-01

    Bacterial endophytes ubiquitously colonize the internal tissues of plants, being found in nearly every plant worldwide. Some endophytes are able to promote the growth of plants. For those strains the mechanisms of plant growth-promotion known to be employed by bacterial endophytes are similar to the mechanisms used by rhizospheric bacteria, e.g., the acquisition of resources needed for plant growth and modulation of plant growth and development. Similar to rhizospheric plant growth-promoting bacteria, endophytic plant growth-promoting bacteria can act to facilitate plant growth in agriculture, horticulture and silviculture as well as in strategies for environmental cleanup (i.e., phytoremediation). Genome comparisons between bacterial endophytes and the genomes of rhizospheric plant growth-promoting bacteria are starting to unveil potential genetic factors involved in an endophytic lifestyle, which should facilitate a better understanding of the functioning of bacterial endophytes. PMID:26805622

  7. Endophytic bacteria associated with growing shoot tips of banana (Musa sp.) cv. Grand Naine and the affinity of endophytes to the host.

    PubMed

    Thomas, Pious; Soly, Thyvalappil A

    2009-11-01

    A cultivation-based assessment of endophytic bacteria present in deep-seated shoot tips of banana suckers was made with a view to generate information on the associated organisms, potential endophytic contaminants in tissue-cultured bananas and to assess if the endophytes shared a beneficial relationship with the host. Plating the tissue homogenate from the central core of suckers showed colony growth on nutrient agar from just 75% and 42% of the 12 stocks during May and November, respectively (average 58%; 6 x 10(3) colony-forming units per gram), yielding diverse organisms belonging to firmicutes (Bacillus, Brevibacillus, Paenibacillus, Virgibacillus, Staphylococcus spp.), actinobacteria (Cellulomonas, Micrococcus, Corynebacterium, Kocuria spp.), alpha-proteobacteria (Paracoccus sp.), and gamma-proteobacteria (Pseudomonas, Acinetobacter spp.). Each shoot tip showed one to three different organisms and no specific organism appeared common to different sucker tips. Tissue homogenate from shoot tips including the ones that did not yield culturable bacteria displayed abundant bacterial cells during microscopic examination suggesting that a high proportion of cells were in viable-but-nonculturable state, or their cultivation requirements were not met. Direct application of cultivation-independent approach to study endophytic bacterial community using bacterial 16S ribosomal RNA universal primers resulted in high interference from chloroplast and mitochondrial genome sequences. Dislodging the bacterial cells from shoot tips that did not show cultivable bacteria and incubating the tissue crush in dilute-nutrient broth led to the activation of four organisms (Klebsiella, Agrobacterium, Pseudacidovorax spp., and an unidentified isolate). The endophytic organisms in general showed better growth at 30-37 degrees C compared with 25 degrees C, and the growth of endophytes as well as pathogenic Erwinia carotovora were promoted with the supply of host tissue extract (HTE) while that of the isolates from nonplant sources were inhibited or unaffected by HTE, suggesting an affinity or dependence of the endophytes on the host and the prospect of an HTE-based assay for discriminating the nonendophytes from endophytes. PMID:19633807

  8. Role of mechanical vs. chemical action in the removal of adherent Bacillus spores during CIP procedures.

    PubMed

    Faille, C; Bénézech, T; Blel, W; Ronse, A; Ronse, G; Clarisse, M; Slomianny, C

    2013-04-01

    This study was designed to evaluate the respective roles of mechanical and chemical effects on the removal of Bacillus spores during cleaning-in-place. This analysis was performed on 12 strains belonging to the Bacillus cereus group (B. cereus, Bacillus anthracis, Bacillus thuringiensis) or to less related Bacillus species (Bacillus pumilus, Bacillus licheniformis, Bacillus sporothermodurans, Bacillus subtilis). Adherent spores were subjected to rinsing-in-place (mechanical action) and cleaning-in-place (mechanical and chemical actions) procedures, the latter involving NaOH 0.5% at 60°C. Results revealed that mechanical action alone only removed between 53 and 89% of the attached spores at a shear stress of 500 Pa. This resistance to shear was not related to spore surface properties. Conversely, in the presence of NaOH at a shear stress of 4 Pa, spores were readily detached, with between 80 and 99% of the adherent spores detached during CIP and the chemical action greatly depended on the strain. This finding suggests that chemical action plays the major role during CIP, whose efficacy is significantly governed by the spore surface chemistry. PMID:23200646

  9. Investigation of the molecular mechanism of thermal tolerance in bacillus subtilis. Final report, August 15, 1980-August 14, 1981

    SciTech Connect

    Yamamoto, N.; Alexander, J.; Ch'ih, J.

    1981-08-14

    We have studied Bacillus subtilis, Bacillus pumilus and Bacillus caldolyticus to ascertain the molecular mechanism of the ability to growth at high temperatures. B. subtilus wild type strains grow in minimal salts medium at a mesophilic temperature (37/sup 0/C) but not at a thermophilic temperature (56/sup 0/C). However, they do grow at 56/sup 0/C in complex rich media. Analysis of their nutritional requirements revealed that these bacteria require pyridoxine or aspartic acid and threonine to grow at 56/sup 0/C indicating that they are temperature sensitive mutants. Furthermore, mutants of B. subtilis which are able to grow on minimal salts media at 56/sup 0/C can be readily isolated. Therefore, it appears that the lack of growth of the wild type strain in minimal salts media at 56/sup 0/C is due to the instability of an anabolic enzyme. In contrast to B. subtilis, B. pumilus has never been observed to growth above 50/sup 0/C. However, our studies show that mutants of B. pumilus and B. subtilis which grow at 68/sup 0/C can be easily isolated. The isolation of such mutants strongly supports the idea that one gene can determine the ability to grow at extreme temperatures.

  10. Diversity of endophytic fungal and bacterial communities in Ilex paraguariensis grown under field conditions.

    PubMed

    Pérez, María Laura; Collavino, Mónica Mariana; Sansberro, Pedro Alfonso; Mroginski, Luis Amado; Galdeano, Ernestina

    2016-04-01

    The composition and diversity of the endophytic community associated with yerba mate (Ilex paraguariensis) was investigated using culture-depending methods. Fungi were identified based on their micromorphological characteristics and internal transcribed spacer rDNA sequence analysis; for bacteria 16S rDNA sequence analysis was used. Fungal and bacterial diversity did not show significant differences between organ age. The highest fungal diversity was registered during fall season and the lowest in winter. Bacterial diversity was higher in stems and increased from summer to winter, in contrast with leaves, which decreased. The most frequently isolated fungus was Fusarium, followed by Colletotrichum; they were both present in all the sampling seasons and organ types assayed. Actinobacteria represented 57.5 % of all bacterial isolates. The most dominant bacterial taxa were Curtobacterium and Microbacterium. Other bacteria frequently found were Methylobacterium, Sphingomonas, Herbiconiux and Bacillus. Nitrogen fixation and phosphate solubilization activity, ACC deaminase production and antagonism against plant fungal pathogens were assayed in endophytic bacterial strains. In the case of fungi, strains of Trichoderma, Penicillium and Aspergillus were assayed for antagonism against pathogenic Fusarium sp. All microbial isolates assayed showed at least one growth promoting activity. Strains of Bacillus, Pantoea, Curtobacterium, Methylobacterium, Brevundimonas and Paenibacillus had at least two growth-promoting activities, and Bacillus, Paenibacillus and the three endophytic fungi showed high antagonistic activity against Fusarium sp. In this work we have made a wide study of the culturable endophytic community within yerba mate plants and found that several microbial isolates could be considered as potential inoculants useful for improving yerba mate production. PMID:26925623

  11. Fungal Endophyte Diversity in Sarracenia

    PubMed Central

    Glenn, Anthony; Bodri, Michael S.

    2012-01-01

    Fungal endophytes were isolated from 4 species of the carnivorous pitcher plant genus Sarracenia: S. minor, S. oreophila, S. purpurea, and S. psittacina. Twelve taxa of fungi, 8 within the Ascomycota and 4 within the Basidiomycota, were identified based on PCR amplification and sequencing of the internal transcribed spacer sequences of nuclear ribosomal DNA (ITS rDNA) with taxonomic identity assigned using the NCBI nucleotide megablast search tool. Endophytes are known to produce a large number of metabolites, some of which may contribute to the protection and survival of the host. We speculate that endophyte-infected Sarracenia may benefit from their fungal associates by their influence on nutrient availability from within pitchers and, possibly, by directly influencing the biota within pitchers. PMID:22427921

  12. Fungal endophyte diversity in Sarracenia.

    PubMed

    Glenn, Anthony; Bodri, Michael S

    2012-01-01

    Fungal endophytes were isolated from 4 species of the carnivorous pitcher plant genus Sarracenia: S. minor, S. oreophila, S. purpurea, and S. psittacina. Twelve taxa of fungi, 8 within the Ascomycota and 4 within the Basidiomycota, were identified based on PCR amplification and sequencing of the internal transcribed spacer sequences of nuclear ribosomal DNA (ITS rDNA) with taxonomic identity assigned using the NCBI nucleotide megablast search tool. Endophytes are known to produce a large number of metabolites, some of which may contribute to the protection and survival of the host. We speculate that endophyte-infected Sarracenia may benefit from their fungal associates by their influence on nutrient availability from within pitchers and, possibly, by directly influencing the biota within pitchers. PMID:22427921

  13. Metabolic potential of endophytic bacteria☆

    PubMed Central

    Brader, Günter; Compant, Stéphane; Mitter, Birgit; Trognitz, Friederike; Sessitsch, Angela

    2014-01-01

    The bacterial endophytic microbiome promotes plant growth and health and beneficial effects are in many cases mediated and characterized by metabolic interactions. Recent advances have been made in regard to metabolite production by plant microsymbionts showing that they may produce a range of different types of metabolites. These substances play a role in defense and competition, but may also be needed for specific interaction and communication with the plant host. Furthermore, few examples of bilateral metabolite production are known and endophytes may modulate plant metabolite synthesis as well. We have just started to understand such metabolic interactions between plants and endophytes, however, further research is needed to more efficiently make use of beneficial plant-microbe interactions and to reduce pathogen infestation as well as to reveal novel bioactive substances of commercial interest. PMID:24863894

  14. Bacterial endophyte communities in the foliage of coast redwood and giant sequoia.

    PubMed

    Carrell, Alyssa A; Frank, Anna C

    2015-01-01

    The endophytic bacterial microbiome, with an emerging role in plant nutrient acquisition and stress tolerance, is much less studied in natural plant populations than in agricultural crops. In a previous study, we found consistent associations between trees in the pine family and acetic acid bacteria (AAB) occurring at high relative abundance inside their needles. Our objective here was to determine if that pattern may be general to conifers, or alternatively, is more likely restricted to pines or conifers growing in nutrient limited and exposed environments. We used 16S rRNA pyrosequencing to characterize the foliar endophyte communities of two conifers in the Cupressaceae family: Two coast redwood (CR; Sequoia sempervirens) populations and one giant sequoia (GS; Sequoiadendron giganteum) population were sampled. Similar to the pines, the endophyte communities of the giant trees were dominated by Proteobacteria, Firmicutes, Acidobacteria, and Actinobacteria. However, although some major operational taxonomic units (OTUs) occurred at a high relative abundance of 10-40% in multiple samples, no specific group of bacteria dominated the endophyte community to the extent previously observed in high-elevation pines. Several of the dominating bacterial groups in the CR and GS foliage (e.g., Bacillus, Burkholderia, Actinomycetes) are known for disease- and pest suppression, raising the possibility that the endophytic microbiome protects the giant trees against biotic stress. Many of the most common and abundant OTUs in our dataset were most similar to 16S rRNA sequences from bacteria found in lichens or arctic plants. For example, an OTU belonging to the uncultured Rhizobiales LAR1 lineage, which is commonly associated with lichens, was observed at high relative abundance in many of the CR samples. The taxa shared between the giant trees, arctic plants, and lichens may be part of a broadly defined endophyte microbiome common to temperate, boreal, and tundra ecosystems. PMID:26441933

  15. Bacterial endophyte communities in the foliage of coast redwood and giant sequoia

    PubMed Central

    Carrell, Alyssa A.; Frank, Anna C.

    2015-01-01

    The endophytic bacterial microbiome, with an emerging role in plant nutrient acquisition and stress tolerance, is much less studied in natural plant populations than in agricultural crops. In a previous study, we found consistent associations between trees in the pine family and acetic acid bacteria (AAB) occurring at high relative abundance inside their needles. Our objective here was to determine if that pattern may be general to conifers, or alternatively, is more likely restricted to pines or conifers growing in nutrient limited and exposed environments. We used 16S rRNA pyrosequencing to characterize the foliar endophyte communities of two conifers in the Cupressaceae family: Two coast redwood (CR; Sequoia sempervirens) populations and one giant sequoia (GS; Sequoiadendron giganteum) population were sampled. Similar to the pines, the endophyte communities of the giant trees were dominated by Proteobacteria, Firmicutes, Acidobacteria, and Actinobacteria. However, although some major operational taxonomic units (OTUs) occurred at a high relative abundance of 10–40% in multiple samples, no specific group of bacteria dominated the endophyte community to the extent previously observed in high-elevation pines. Several of the dominating bacterial groups in the CR and GS foliage (e.g., Bacillus, Burkholderia, Actinomycetes) are known for disease- and pest suppression, raising the possibility that the endophytic microbiome protects the giant trees against biotic stress. Many of the most common and abundant OTUs in our dataset were most similar to 16S rRNA sequences from bacteria found in lichens or arctic plants. For example, an OTU belonging to the uncultured Rhizobiales LAR1 lineage, which is commonly associated with lichens, was observed at high relative abundance in many of the CR samples. The taxa shared between the giant trees, arctic plants, and lichens may be part of a broadly defined endophyte microbiome common to temperate, boreal, and tundra ecosystems. PMID:26441933

  16. Monitoring the ecology of Bacillus during Daqu incubation, a fermentation starter, using culture-dependent and culture-independent methods.

    PubMed

    Yan, Zheng; Zheng, Xiao-Wei; Han, Bei-Zhong; Han, Jian-Shu; Nout, M J Robert; Chen, Jing-Yu

    2013-05-01

    Daqu, a traditional fermentation starter, has been used to produce attractively flavored foods such as vinegar and Chinese liquor for thousands of years. Although Bacillus spp. are one of the dominant microorganisms in Daqu, more precise information is needed to reveal why and how Bacillus became dominant in Daqu, and next, to assess the impact of Bacillus sp. on Daqu and its derived products. We combined culture-dependent and culture-independent methods to study the ecology of Bacillus during Daqu incubation. Throughout the incubation, 67 presumptive Bacillus spp. isolates were obtained, 52 of which were confirmed by 16S rDNA sequencing. The identified organisms belonged to 8 Bacillus species: B. licheniformis, B. subtilis, B. amyloliquefaciens, B. cereus, B. circulans, B. megaterium, B. pumilus, and B. anthracis. A primer set specific for Bacillus and related genera was used in a selective PCR study, followed by a nested DGGE PCR targeting the V9 region of the 16S rDNA. Species identified from the PCR-DGGE fingerprints were related to B. licheniformis, B. subtilis, B. amyloliquefaciens, B. pumilus, B. benzoevorans, and B. foraminis. The predominant species was found to be B. licheniformis. Certain B. licheniformis strains exhibited potent antimicrobial activities. The greatest species diversity occurred at the Liangmei stage of Daqu incubation. To date, we lack sufficient knowledge of Bacillus distribution in Daqu. Elucidating the ecology of Bacillus during Daqu incubation would enable the impact of Bacillus on Daqu to be accessed, and the quality and stabilization of Daqu-derived products to be optimized. PMID:23648849

  17. Plant-endophyte-herbivore interactions

    PubMed Central

    Parsons, Anthony J; Popay, Alison; Xue, Hong; Newman, Jonathan A

    2008-01-01

    A recent paper by Rasmussen et al., (New Phytol 2007; 173:787–97) describes the interactions between Lolium perenne cultivars with contrasting carbohydrate content and the symbiotic fungal endophyte Neotyphodium lolii at different levels of nitrogen supply. In a subsequent study undertaken by Rasmussen et al., (Plant Physiol 2008; 146:1440–53) 66 metabolic variables were analysed in the same material, revealing widespread effects of endophyte infection, N supply and cultivar carbohydrate content on both primary and secondary metabolites. Here, we link insect numerical responses to these metabolic responses using multiple regression analysis. PMID:19704424

  18. Sterilization of Bacillus spores by converted X rays

    NASA Astrophysics Data System (ADS)

    Ito, Hitoshi; Ohki, Yumi; Watanabe, Yuhei; Sunaga, Hiromi; Ishigaki, Isao

    1993-10-01

    Relative sensitivities of endospores of Bacillus pumilus E601, B. subtilis IAM1069, B. megaterium S31 and B. brevis S5 to gamma rays, converted X rays (bremsstrahlung) and electron beams were examined in order to estimate the conditions in which converted X rays kill Bacillus spores. The radiation sensitivities to gamma rays and electron beams of each strain dried on glass fiber filter without additives were found to be almost equivalent, and D values were obtained as follows: 1.5-1.6 kGy for B. pumilus, 1.4-1.5 kGy for B. subtilis, 1.9-2.0 kGy for B. megaterium and 1.6-2.0 kGy for B. brevis. The radiation sensitivities of endospores of each strain to electron beams were slightly lower than those to gamma rays in the dry condition with additives of 2% peptone + 1 % glycerin on glass fiber filters. The increase of radiation resistance in the presence of additives was also observed with X rays, and it was on an intermediate level between those with gamma rays and electron beams. In the dry condition using cellulose filter paper, only the radiation resistances of B. megaterium and B. brevis in the presence of additives B. megaterium and B. brevis in the presence of additives were increased.

  19. MICROARRAY COMPARISON OF TALL FESCUE GENE EXPRESSION IN ENDOPHYTE INFECTED AND ENDOPHYTE FREE PLANTS

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Many grasses have mutualistic symbioses with fungi of the family Clavicipitaceae. Tall fescue (Lolium arundinaceum) can harbor the obligate endophyte, Neotyphodium coenophialum that are asexually propagated and transmitted via host seeds. The endophyte receives shelter and nutrients from the host ...

  20. Diversity of bacteria of the genus Bacillus on board of international space station.

    PubMed

    Alekhova, T A; Zakharchuk, L M; Tatarinova, N Yu; Kadnikov, V V; Mardanov, A V; Ravin, N V; Skryabin, K G

    2015-11-01

    From swabs of surfaces of equipment and air samples of the Russian segment of the International Space Station, nine strains of spore-forming bacteria of the genus Bacillus belonging to the species B. pumilus, B. licheniformis, B. subtilis, B. megaterium, and B. amyloliquefaciens were isolated. The last species of bacilli on the equipment of RS ISS was detected for the first time. For these species of bacilli, there are known strains that can be opportunistic to humans, and their metabolites can cause biodegradation of equipment and materials. B. pumilus found on ISS belongs to the group of bacteria that exhibits a particularly high resistance to adverse environmental conditions, such as dehydration, ultraviolet and gamma radiation, and chemical disinfection. PMID:26728721

  1. Effects of growth stage and fulvic acid on the diversity and dynamics of endophytic bacterial community in Stevia rebaudiana Bertoni leaves

    PubMed Central

    Yu, Xuejian; Yang, Jinshui; Wang, Entao; Li, Baozhen; Yuan, Hongli

    2015-01-01

    The aim of this study was to learn the interactions among the endophytic bacteria, the plant growth, the foliar spray of fulvic acid, and the accumulation of steviol glycosides in the leaves of Stevia rebaudiana. Metagenomic DNA was extracted from the Stevia leaves at different growth stages with or without the fulvic acid treatment; and the diversity of endophytic bacteria in Stevia leaves was estimated by pyrosequencing of 16S rRNA genes. As results, Proteobacteria, Actinobacteria, Bacteroidetes, and Firmicutes were found to be the dominant phyla despite the growth stages and fulvic acid application. Stevia growth stages strongly regulated composition of endophytic community. The genera Agrobacterium (12.3%) and Erwinia (7.2%) dominated in seedling stage were apparently declined in the vegetable and initial flowering stages, while Sphingomonas and Methylobacterium increased in mature leaves at harvest time, which showed that the mature leaves of Stevia preferred to accumulate some certain endophytic bacteria. Sphingomonas and Methylobacterium constituted an important part of the core endophytic community and were positively correlated with the stevioside content and UGT74G1 gene expression, respectively; while Erwinia, Agrobacterium, and Bacillus were negatively correlated with the stevioside accumulation. Fulvic acid treatment accelerated the variation of endophytes along the growth stages and increased the steviol glycosides content. This is the first study to reveal the community composition of endophytic bacteria in the Stevia leaves, to evidence the strong effects of growth stage and fulvic acid application on the endophytes of Stevia, and to demonstrate the correlation between the endophytic bacteria and the steviol glycosides accumulation. PMID:26379644

  2. Effects of growth stage and fulvic acid on the diversity and dynamics of endophytic bacterial community in Stevia rebaudiana Bertoni leaves.

    PubMed

    Yu, Xuejian; Yang, Jinshui; Wang, Entao; Li, Baozhen; Yuan, Hongli

    2015-01-01

    The aim of this study was to learn the interactions among the endophytic bacteria, the plant growth, the foliar spray of fulvic acid, and the accumulation of steviol glycosides in the leaves of Stevia rebaudiana. Metagenomic DNA was extracted from the Stevia leaves at different growth stages with or without the fulvic acid treatment; and the diversity of endophytic bacteria in Stevia leaves was estimated by pyrosequencing of 16S rRNA genes. As results, Proteobacteria, Actinobacteria, Bacteroidetes, and Firmicutes were found to be the dominant phyla despite the growth stages and fulvic acid application. Stevia growth stages strongly regulated composition of endophytic community. The genera Agrobacterium (12.3%) and Erwinia (7.2%) dominated in seedling stage were apparently declined in the vegetable and initial flowering stages, while Sphingomonas and Methylobacterium increased in mature leaves at harvest time, which showed that the mature leaves of Stevia preferred to accumulate some certain endophytic bacteria. Sphingomonas and Methylobacterium constituted an important part of the core endophytic community and were positively correlated with the stevioside content and UGT74G1 gene expression, respectively; while Erwinia, Agrobacterium, and Bacillus were negatively correlated with the stevioside accumulation. Fulvic acid treatment accelerated the variation of endophytes along the growth stages and increased the steviol glycosides content. This is the first study to reveal the community composition of endophytic bacteria in the Stevia leaves, to evidence the strong effects of growth stage and fulvic acid application on the endophytes of Stevia, and to demonstrate the correlation between the endophytic bacteria and the steviol glycosides accumulation. PMID:26379644

  3. Morphological and genetic characterization of endophytic bacteria isolated from roots of different maize genotypes.

    PubMed

    Ikeda, Angela Cristina; Bassani, Luciana Lange; Adamoski, Douglas; Stringari, Danyelle; Cordeiro, Vanessa Kava; Glienke, Chirlei; Steffens, Maria Berenice Reynaud; Hungria, Mariangela; Galli-Terasawa, Lygia Vitoria

    2013-01-01

    Maize is one of the most important crops worldwide, and in Brazil, the state of Paraná stands as its largest producer. The crop demands high inputs of N fertilizers, therefore all strategies aiming to optimize the grain production with lower inputs are very relevant. Endophytic bacteria have a high potential to increment maize grain yield by means of input via biological nitrogen fixation and/or plant growth promotion, in this last case increasing the absorption of water and nutrients by the plants. In this study, we established a collection of 217 endophytic bacteria, isolated from roots of four lineages and three hybrid genotypes of maize, and isolated in four different N-free culture media. Biochemical-comprising growth in different carbon sources, intrinsic tolerance to antibiotics, and biochemical tests for catalase, nitrate reductase, urease, and growth in N-free media in vitro-and genetic characterization by BOX-PCR revealed great variability among the isolates. Both commercial hybrids and homozygous lineages were broadly colonized by endophytes, and sequencing of the 16S rRNA gene revealed the presence of bacteria belonging to the genera Pantoea, Bacillus, Burkholderia, and Klebsiella. Qualitative differences in endophytic colonization were detected between lineages and hybrid genotypes. PMID:22956211

  4. WHAT DOES AN ENDOPHYTE LOOK LIKE? ENDOPHYTE PROFILES OF NATIVE GRASSES AND SHRUBS

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Obligate fungal endophytes often defy detection and isolation by conventional microbiology techniques. Staining methods, including the use of trypan blue and sudan IV have effectively detected endophytes in plant tissues. However, stains fail to distinguish one fungal endophyte from another, thus ...

  5. Endophyte-associated ergot alkaloids

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Fescue toxicosis is a very costly (greater than $600 million/annually) for the cattle, horse and small ruminant industries. The tall fescue forage responsible for this intoxication is infected with an endophytic fungus (Neotyphodium coenophialum) that produces ergot alkaloids, which are toxic to th...

  6. Fungal endophyte diversity in Sarracenia

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Fungal endophytes were isolated from four species of the carnivorous pitcher plant genus Sarracenia: S. minor, S. oreophila, S. purpurea, and S. psittacina. Twelve taxa of fungi, eight within the Ascomycota and four within the Basidiomycota, were identified based on PCR amplification and sequencing ...

  7. Antimicrobial activity of Ulva reticulata and its endophytes

    NASA Astrophysics Data System (ADS)

    Dhanya, K. I.; Swati, V. I.; Vanka, Kanth Swaroop; Osborne, W. J.

    2016-04-01

    Seaweeds are known to exhibit various antimicrobial properties, since it harbours an enormous range of indigenous bioactive compounds. The emergence of drug resistant strains has directed to the identification of prospective metabolites from seaweed and its endophytes, thereby exploiting the properties in resisting bacterial diseases. The current study was aimed to assess the antimicrobial activity of extracts obtained from Ulva reticulate, for which metabolites of Ulva reticulata and its endophytes were extracted and assessed against human pathogens like Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Salmonella typhi, and Bacillus subtilis. It was observed that the hexane extract of isolate VITDSJ2 was effective against all the tested pathogens but a significant inhibition was observed for Staphylococcus aureus and Escherichia coli. Further, Gas chromatography coupled with Mass spectroscopy (GC-MS) revealed the existence of phenol, 3, 5-bis (1, 1-dimethylethyl) in the crude hexane extract which is well-known to possess antibacterial activity. The effective isolate VITDSJ2 was identified to be the closest neighbour of Pseudomonas stutzeri by phenotypic and genotypic methods. The crude extracts of the seaweed Ulva reticulata was also screened for antibacterial activity and the hexane extract was effective in showing inhibition against all the tested pathogens. The compound in the crude extract of Ulva reticulata was identified as hentriacontane using GC-MS. The extracts obtained from dichloromethane did not show significant activity in comparison with the hexane extracts. Hence the metabolites of Ulva reticulata and the bacterial secondary metabolites of the endophytes could be used in the treatment of bacterial infections.

  8. Chitin extraction from crab shells by Bacillus bacteria. Biological activities of fermented crab supernatants.

    PubMed

    Hajji, Sawssen; Ghorbel-Bellaaj, Olfa; Younes, Islem; Jellouli, Kemel; Nasri, Moncef

    2015-08-01

    Crab shells waste were fermented using six protease-producing Bacillus species (Bacillus subtilis A26, Bacillus mojavensis A21, Bacillus pumilus A1, Bacillus amyloliquefaciens An6, Bacillus licheniformis NH1 and Bacillus cereus BG1) for the production of chitin and fermented-crab supernatants (FCSs). In medium containing only crab shells, the highest demineralization DM was obtained with B. licheniformis NH1 (83±0.5%) and B. pumilus A1 (80±0.6%), while the highest deproteinization (DP) was achieved with A1 (94±1%) followed by NH1 (90±1.5%) strains. Cultures conducted in medium containing crab shells waste supplemented with 5% (w/v) glucose, were found to remarkably promote demineralization efficiency, and enhance slightly deproteinization rates. FTIR spectra of chitins showed the characteristics bands of ?-chitin. FCSs showed varying degrees of antioxidant activities which were in a dose-dependent manner (p<0.01). In fact, FCS produced by B. amyloliquefaciens An6 exhibited the highest DPPH free radical-scavenging activity (92% at 4 mg/ml), while the lowest hydroxyl radical-scavenging activity (60% at 4 mg/ml) was obtained with B. subtilis A26 hydrolysates. However, the highest reducing power (OD700nm=2 at 0.5 mg/ml) was obtained by B.amyloliquefaciens An6 hydrolysates. These results suggest that crab hydrolysates are good sources of natural antioxidants. Further, FCSs were found to exhibit antibacterial activity against Gram-positive and Gram-negative bacteria. PMID:25910648

  9. Endophytic bacterial community living in roots of healthy and 'Candidatus Phytoplasma mali'-infected apple (Malus domestica, Borkh.) trees.

    PubMed

    Bulgari, Daniela; Bozkurt, Adem I; Casati, Paola; Ca?layan, Kadriye; Quaglino, Fabio; Bianco, Piero A

    2012-11-01

    'Candidatus Phytoplasma mali', the causal agent of apple proliferation (AP) disease, is a quarantine pathogen controlled by chemical treatments against insect vectors and eradication of diseased plants. In accordance with the European Community guidelines, novel strategies should be developed for sustainable management of plant diseases by using resistance inducers (e.g. endophytes). A basic point for the success of this approach is the study of endophytic bacteria associated with plants. In the present work, endophytic bacteria living in healthy and 'Ca. Phytoplasma mali'-infected apple trees were described by cultivation-dependent and independent methods. 16S rDNA sequence analysis showed the presence of the groups Proteobacteria, Acidobacteria, Bacteroidetes, Actinobacteria, Chlamydiae, and Firmicutes. In detail, library analyses underscored 24 and 17 operational taxonomic units (OTUs) in healthy and infected roots, respectively, with a dominance of Betaproteobacteria. Moreover, differences in OTUs number and in CFU/g suggested that phytoplasmas could modify the composition of endophytic bacterial communities associated with infected plants. Intriguingly, the combination of culturing methods and cloning analysis allowed the identification of endophytic bacteria (e.g. Bacillus, Pseudomonas, and Burkholderia) that have been reported as biocontrol agents. Future research will investigate the capability of these bacteria to control 'Ca. Phytoplasma mali' in order to develop sustainable approaches for managing AP. PMID:22752594

  10. Characterization of endophytic bacteria from cucurbit fruits with potential benefits to agriculture in melons (Cucumis melo L.).

    PubMed

    Glassner, Hanoch; Zchori-Fein, Einat; Compant, Stéphane; Sessitsch, Angela; Katzir, Nurit; Portnoy, Vitaly; Yaron, Sima

    2015-07-01

    Endophytes are microorganisms that mainly colonize vegetative parts, but are also found in reproductive and disseminating organs, and may have beneficial characteristics. To identify microorganisms associated with the agriculturally important family, Cucurbitaceae, endophytes were initially determined in fruits of Cucumis melo Reticulatus Group 'Dulce' by a cultivation-independent approach based on fluorescence in situ hybridization using double labeling of oligonucleotide probes. Alpha-, Beta-, Gammaproteobacteria, Firmicutes and Actinobacteria were localized inside the fruits. Culturable bacteria were further isolated and identified from fruit tissues of 'Dulce', from fruits of other cultivated and wild-field-grown Cucurbitaceae, and from wild fruits growing under natural conditions. Low densities of culturable bacteria were detected in the investigated fruits, especially in four out of the five wild species, regardless of their growing environment. Substantial differences were observed between the wild and cultivated cucurbit taxa in regard to the number of colonized fruits as well as the type of endophytes. Bacillus was the most dominant genus of endophytes colonizing fruits of Cucurbitaceae. The antagonistic effects of isolated endophytes were assessed against cucurbit disease agents in dual-culture assays. Several bacterial isolates exhibited antagonistic properties against the tested plant pathogens. The identified bacteria may be useful for protecting plants not only in the field, but also for post-harvest. PMID:26183916

  11. Endophytes of grapevine flowers, berries, and seeds: identification of cultivable bacteria, comparison with other plant parts, and visualization of niches of colonization.

    PubMed

    Compant, Stéphane; Mitter, Birgit; Colli-Mull, Juan Gualberto; Gangl, Helmut; Sessitsch, Angela

    2011-07-01

    Endophytic bacteria can colonize various plants and organs. However, endophytes colonizing plant reproductive organs have been rarely analyzed. In this study, endophytes colonizing flowers as well as berries and seeds of grapevine plants grown under natural conditions were investigated by cultivation as well as by fluorescence in situ hybridization. For comparison, bacteria were additionally isolated from other plant parts and the rhizosphere and characterized. Flowers, fruits, and seeds hosted various endophytic bacteria. Some taxa were specifically isolated from plant reproductive organs, whereas others were also detected in the rhizosphere, endorhiza or grape inflo/infructescence stalk at the flowering or berry harvest stage. Microscopic analysis by fluorescence in situ hybridization of resin-embedded samples confirmed the presence of the isolated taxa in plant reproductive organs and enabled us to localize them within the plant. Gammaproteobacteria (including Pseudomonas spp.) and Firmicutes (including Bacillus spp.) were visualized inside the epidermis and xylem of ovary and/or inside flower ovules. Firmicutes, mainly Bacillus spp. were additionally visualized inside berries, in the intercellular spaces of pulp cells and/or xylem of pulp, but also along some cell walls inside parts of seeds. Analysis of cultivable bacteria as well as microscopic results indicated that certain endophytic bacteria can colonize flowers, berries, or seeds. Our results also indicated that some specific taxa may not only derive from the root environment but also from other sources such as the anthosphere. PMID:21625971

  12. [Stress resistance and genetic diversity of endophytic bacteria isolated from Caragana spp. root nodules].

    PubMed

    Dai, Jin-Xia; Wang, Yu-Jiong; Wu, Xue-Juan; Zhang, Xiao-Ling

    2012-02-01

    By adopting PCR-RFLP and 16S rDNA sequencing, this paper analyzed the genetic diversity and phylogeny of 40 endophytic bacterial strains isolated from Caragana spp. root nodules, and determined the salt resistance, acid- and alkali resistance, and growth temperature range of the strains. A total of 9 genotypes were obtained from the 40 strains by RFLP. The 16S rDNA sequencing, morphological observation, and biochemical test of representative strains showed that the strains belonged to Bacillus, Inquilinus, Shinella and Acinetobacter, respectively, and had rich genetic diversity. 57.5% of the strains could tolerate 4% NaCl stress, 75% of the strains could grow in YMA medium with an initial pH 11.0, and 85% of the strains could survive after heat shock treatment at 60 degrees C, suggesting that the endophytic bacteria of Caragana spp. had strong resistance capacity. Among the strains, LWEN 07 and LWEN 15 were most resistant. PMID:22586981

  13. In silico analysis of the 16S rRNA gene of endophytic bacteria, isolated from the aerial parts and seeds of important agricultural crops.

    PubMed

    Bredow, C; Azevedo, J L; Pamphile, J A; Mangolin, C A; Rhoden, S A

    2015-01-01

    Because of human population growth, increased food production and alternatives to conventional methods of biocontrol and development of plants such as the use of endophytic bacteria and fungi are required. One of the methods used to study microorganism diversity is sequencing of the 16S rRNA gene, which has several advantages, including universality, size, and availability of databases for comparison. The objective of this study was to analyze endophytic bacterial diversity in agricultural crops using published papers, sequence databases, and phylogenetic analysis. Fourteen papers were selected in which the ribosomal 16S rRNA gene was used to identify endophytic bacteria, in important agricultural crops, such as coffee, sugar cane, beans, corn, soybean, tomatoes, and grapes, located in different geographical regions (America, Europe, and Asia). The corresponding 16S rRNA gene sequences were selected from the NCBI database, aligned using the Mega 5.2 program, and phylogenetic analysis was undertaken. The most common orders present in the analyzed cultures were Bacillales, Enterobacteriales, and Actinomycetales and the most frequently observed genera were Bacillus, Pseudomonas, and Microbacterium. Phylogenetic analysis showed that only approximately 1.56% of the total sequences were not properly grouped, demonstrating reliability in the identification of microorganisms. This study identified the main genera found in endophytic bacterial cultures from plants, providing data for future studies on improving plant agriculture, biotechnology, endophytic bacterium prospecting, and to help understand relationships between endophytic bacteria and their interactions with plants. PMID:26345903

  14. Isolation of endophytic bacteria from arboreal species of the Amazon and identification by sequencing of the 16S rRNA encoding gene

    PubMed Central

    Coêlho, Mariza M.; Ferreira-Nozawa, Monica S.; Nozawa, Sérgio R.; Santos, André L.W.

    2011-01-01

    Endophytic bacteria from three arboreal species native to the Amazon (Carapa guianenses, Ceiba pentandra, and Swietenia macrophylla), were isolated and identified, through partial sequencing of the 16S rRNA encoding gene. From these, 16 isolates were obtained, although, when compared to sequences deposited in GenBank, only seven had produced identifiable fragments. Bacillus, Pantoea and two non-culturable samples were identified. Results obtained through sequence analysis revealed low genetic diversity across the isolates, even when analyzing different species and plant structures. This is the first report concerning the isolation and identification of endophytic bacteria in these plant species. PMID:22215973

  15. Isolation of endophytic bacteria from arboreal species of the Amazon and identification by sequencing of the 16S rRNA encoding gene.

    PubMed

    Coêlho, Mariza M; Ferreira-Nozawa, Monica S; Nozawa, Sérgio R; Santos, André L W

    2011-10-01

    Endophytic bacteria from three arboreal species native to the Amazon (Carapa guianenses, Ceiba pentandra, and Swietenia macrophylla), were isolated and identified, through partial sequencing of the 16S rRNA encoding gene. From these, 16 isolates were obtained, although, when compared to sequences deposited in GenBank, only seven had produced identifiable fragments. Bacillus, Pantoea and two non-culturable samples were identified. Results obtained through sequence analysis revealed low genetic diversity across the isolates, even when analyzing different species and plant structures. This is the first report concerning the isolation and identification of endophytic bacteria in these plant species. PMID:22215973

  16. Identification of single nucleotide polymorphisms (SNPs) in the 16S rRNA gene of foodborne Bacillus spp.

    PubMed

    Fernández-No, I C; Böhme, K; Caamaño-Antelo, S; Barros-Velázquez, J; Calo-Mata, P

    2015-04-01

    The main goal of this work was the identification of single nucleotide polymorphisms (SNPs) in the 16S rRNA gene of foodborne Bacillus spp. that may be useful for typing purposes. These species include, among others, Bacillus cereus, an important pathogenic species involved in food poisoning, and Bacillus licheniformis, Bacillus subtilis and Bacillus pumilus, which are causative agents of food spoilage described as responsible for foodborne disease outbreaks. With this purpose in mind, 52 Bacillus strains isolated from culture collections and fresh and processed food were considered. SNP type "Y" at sites 212 and 476 appeared in the majority of B. licheniformis studied strains. SNP type "R" at site 278 was detected in many strains of the B. subtilis/Bacillus amyloliquefaciens group, while polymorphism "Y" at site 173 was characteristic of the majority of strains of B. cereus/Bacillus thuringiensis group. The analysis of SNPs provided more intra-specific information than phylogenetic analysis in the cases of B. cereus and B. subtilis. Moreover, this study describes novel SNPs that should be considered when designing 16S rRNA-based primers and probes for multiplex-PCR, Real-Time PCR and microarray systems for foodborne Bacillus spp. PMID:25475292

  17. Isolation and characterization of endophytic bacteria isolated from the leaves of the common bean (Phaseolus vulgaris)

    PubMed Central

    de Oliveira Costa, Leonardo Emanuel; de Queiroz, Marisa Vieira; Borges, Arnaldo Chaer; de Moraes, Celia Alencar; de Araújo, Elza Fernandes

    2012-01-01

    The common bean is one of the most important legumes in the human diet, but little is known about the endophytic bacteria associated with the leaves of this plant. The objective of this study was to characterize the culturable endophytic bacteria of common bean (Phaseolus vulgaris) leaves from three different cultivars (Vermelhinho, Talismã, and Ouro Negro) grown under the same field conditions. The density of endophytic populations varied from 4.5 x 102 to 2.8 x 103 CFU g-1 of fresh weight. Of the 158 total isolates, 36.7% belonged to the Proteobacteria, 32.9% to Firmicutes, 29.7% to Actinobacteria, and 0.6% to Bacteroidetes. The three P. vulgaris cultivars showed class distribution differences among Actinobacteria, Alphaproteobacteria and Bacilli. Based on 16S rDNA sequences, 23 different genera were isolated comprising bacteria commonly associated with soil and plants. The genera Bacillus, Delftia, Methylobacterium, Microbacterium, Paenibacillus, Staphylococcus and Stenotrophomonas were isolated from all three cultivars. To access and compare the community structure, diversity indices were calculated. The isolates from the Talismã cultivar were less diverse than the isolates derived from the other two cultivars. The results of this work indicate that the cultivar of the plant may contribute to the structure of the endophytic community associated with the common bean. This is the first report of endophytic bacteria from the leaves of P. vulgaris cultivars. Future studies will determine the potential application of these isolates in biological control, growth promotion and enzyme production for biotechnology. PMID:24031988

  18. Surfactin A production and isoforms characterizations in strains of Bacillus mojavensis for control of a maize pathogen, Fusarium verticillioides

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The endophytic bacterium, Bacillus mojavensis, RRC 101 controls fungal diseases in maize and other plants. The bacterium and its cultural extracts have been shown to be antagonistic to the pathogenic and mycotoxic fungus, Fusarium verticillioides. An antifungal cyclic lipopeptide produced by B. moj...

  19. Impact of the Endophyte on Animal Production

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Tall fescue (Lolium arundinaceum L.) is extensively utilized as pasture forage in the upper transition zone of the USA. Its production and persistence is attributed to alkaloids produced by a fungal endophyte (Neotyphodium coenophialum) that infects most tall fescue plants. The endophyte also prod...

  20. Bacterial endophytes: recent developments and applications.

    PubMed

    Ryan, Robert P; Germaine, Kieran; Franks, Ashley; Ryan, David J; Dowling, David N

    2008-01-01

    Endophytic bacteria have been found in virtually every plant studied, where they colonize the internal tissues of their host plant and can form a range of different relationships including symbiotic, mutualistic, commensalistic and trophobiotic. Most endophytes appear to originate from the rhizosphere or phyllosphere; however, some may be transmitted through the seed. Endophytic bacteria can promote plant growth and yield and can act as biocontrol agents. Endophytes can also be beneficial to their host by producing a range of natural products that could be harnessed for potential use in medicine, agriculture or industry. In addition, it has been shown that they have the potential to remove soil contaminants by enhancing phytoremediation and may play a role in soil fertility through phosphate solubilization and nitrogen fixation. There is increasing interest in developing the potential biotechnological applications of endophytes for improving phytoremediation and the sustainable production of nonfood crops for biomass and biofuel production. PMID:18034833

  1. Identification of Bacillus strains by MALDI TOF MS using geometric approach.

    PubMed

    Starostin, Konstantin V; Demidov, Evgeny A; Bryanskaya, Alla V; Efimov, Vadim M; Rozanov, Alexey S; Peltek, Sergey E

    2015-01-01

    Microorganism identification by MALDI TOF mass-spectrometry is based on the comparison of the mass spectrum of the studied organism with those of reference strains. It is a rapid and reliable method. However, commercial databases and programs are mostly designed for identification of clinically important strains and can be used only for particular mass spectrometer models. The need for open platforms and reference databases is obvious. In this study we describe a geometric approach for microorganism identification by mass spectra and demonstrate its capabilities by analyzing 24 strains belonging to the Bacillus pumilus group. This method is based on representing mass spectra as points on a multidimensional space, which allows us to use geometric distances to compare the spectra. Delimitation of microorganisms performed by geometric approach correlates well with the results of molecular phylogenetic analysis and clustering using Biotyper 3.1. All three methods used allowed us to reliably divide the strains into two groups corresponding to closely related species, Bacillus pumilus and Bacillus altitudinis. The method developed by us will be implemented in a Web interface designed for using open reference databases for microorganism identification. The data is available at http://www.bionet.nsc.ru/mbl/database/database.html. PMID:26592761

  2. Identification of Bacillus strains by MALDI TOF MS using geometric approach

    NASA Astrophysics Data System (ADS)

    Starostin, Konstantin V.; Demidov, Evgeny A.; Bryanskaya, Alla V.; Efimov, Vadim M.; Rozanov, Alexey S.; Peltek, Sergey E.

    2015-11-01

    Microorganism identification by MALDI TOF mass-spectrometry is based on the comparison of the mass spectrum of the studied organism with those of reference strains. It is a rapid and reliable method. However, commercial databases and programs are mostly designed for identification of clinically important strains and can be used only for particular mass spectrometer models. The need for open platforms and reference databases is obvious. In this study we describe a geometric approach for microorganism identification by mass spectra and demonstrate its capabilities by analyzing 24 strains belonging to the Bacillus pumilus group. This method is based on representing mass spectra as points on a multidimensional space, which allows us to use geometric distances to compare the spectra. Delimitation of microorganisms performed by geometric approach correlates well with the results of molecular phylogenetic analysis and clustering using Biotyper 3.1. All three methods used allowed us to reliably divide the strains into two groups corresponding to closely related species, Bacillus pumilus and Bacillus altitudinis. The method developed by us will be implemented in a Web interface designed for using open reference databases for microorganism identification. The data is available at http://www.bionet.nsc.ru/mbl/database/database.html.

  3. Identification of Bacillus strains by MALDI TOF MS using geometric approach

    PubMed Central

    Starostin, Konstantin V.; Demidov, Evgeny A.; Bryanskaya, Alla V.; Efimov, Vadim M.; Rozanov, Alexey S.; Peltek, Sergey E.

    2015-01-01

    Microorganism identification by MALDI TOF mass-spectrometry is based on the comparison of the mass spectrum of the studied organism with those of reference strains. It is a rapid and reliable method. However, commercial databases and programs are mostly designed for identification of clinically important strains and can be used only for particular mass spectrometer models. The need for open platforms and reference databases is obvious. In this study we describe a geometric approach for microorganism identification by mass spectra and demonstrate its capabilities by analyzing 24 strains belonging to the Bacillus pumilus group. This method is based on representing mass spectra as points on a multidimensional space, which allows us to use geometric distances to compare the spectra. Delimitation of microorganisms performed by geometric approach correlates well with the results of molecular phylogenetic analysis and clustering using Biotyper 3.1. All three methods used allowed us to reliably divide the strains into two groups corresponding to closely related species, Bacillus pumilus and Bacillus altitudinis. The method developed by us will be implemented in a Web interface designed for using open reference databases for microorganism identification. The data is available at http://www.bionet.nsc.ru/mbl/database/database.html. PMID:26592761

  4. Four new tetramic acid and one new furanone derivatives from the plant endophytic fungus Neopestalotiopsis sp.

    PubMed

    Zhao, Shasha; Chen, Shenxi; Wang, Bo; Niu, Shubin; Wu, Wenping; Guo, Liangdong; Che, Yongsheng

    2015-06-01

    Four new tetramic acid analogues neopestalotins A-D (1-4), one new furanone derivative neopestalotin E (6), and the known compound hymenosetin have been isolated from the solid cultures of the plant endophytic fungus Neopestalotiopsis sp. The structures of the new compounds were determined mainly by nuclear magnetic resonance (NMR) experiments. The absolute configurations of 1 and 2 were assigned by circular dichroism (CD) data, whereas those of 3 and 4 were deduced by a combination of CD and heteronuclear long range coupling (HETLOC) data. Compound 2 showed modest antimicrobial activity against the Gram-positive bacteria, Bacillus subtilis, Staphylococcus aureus col, and Streptococcus pneumoniae. PMID:25818228

  5. Evaluation of the Efficacy and Safety of a Marine-Derived Bacillus Strain for Use as an In-Feed Probiotic for Newly Weaned Pigs

    PubMed Central

    Prieto, Maria Luz; O'Sullivan, Laurie; Tan, Shiau Pin; McLoughlin, Peter; Hughes, Helen; O'Donovan, Orla; Rea, Mary C.; Kent, Robert M.; Cassidy, Joseph P.; Gardiner, Gillian E.; Lawlor, Peadar G.

    2014-01-01

    Forty eight individual pigs (8.7±0.26 kg) weaned at 28±1 d of age were used in a 22-d study to evaluate the effect of oral administration of a Bacillus pumilus spore suspension on growth performance and health indicators. Treatments (n = 16) were: (1) non-medicated diet; (2) medicated diet with apramycin (200 mg/kg) and pharmacological levels of zinc oxide (2,500 mg zinc/kg) and (3) B. pumilus diet (non-medicated diet + 1010 spores/day B. pumilus). Final body weight and average daily gain tended to be lower (P = 0.07) and feed conversion ratio was worsened (P<0.05) for the medicated treatment compared to the B. pumilus treatment. Ileal E. coli counts were lower for the B. pumilus and medicated treatments compared to the non-medicated treatment (P<0.05), perhaps as a result of increased ileal propionic acid concentrations (P<0.001). However, the medicated treatment reduced fecal (P<0.001) and cecal (P<0.05) Lactobacillus counts and tended to reduce the total cecal short chain fatty acid (SCFA) concentration (P = 0.10). Liver weights were lighter and concentrations of liver enzymes higher (P<0.05) in pigs on the medicated treatment compared to those on the non-medicated or B. pumilus treatments. Pigs on the B. pumilus treatment had lower overall lymphocyte and higher granulocyte percentages (P<0.001) and higher numbers of jejunal goblet cells (P<0.01) than pigs on either of the other two treatments or the non-medicated treatment, respectively. However, histopathological examination of the small intestine, kidneys and liver revealed no abnormalities. Overall, the B. pumilus treatment decreased ileal E. coli counts in a manner similar to the medicated treatment but without the adverse effects on growth performance, Lactobacillus counts, cecal SCFA concentration and possible liver toxicity experienced with the medicated treatment. PMID:24586349

  6. Evaluation of the efficacy and safety of a marine-derived Bacillus strain for use as an in-feed probiotic for newly weaned pigs.

    PubMed

    Prieto, Maria Luz; O'Sullivan, Laurie; Tan, Shiau Pin; McLoughlin, Peter; Hughes, Helen; O'Donovan, Orla; Rea, Mary C; Kent, Robert M; Cassidy, Joseph P; Gardiner, Gillian E; Lawlor, Peadar G

    2014-01-01

    Forty eight individual pigs (8.7±0.26 kg) weaned at 28±1 d of age were used in a 22-d study to evaluate the effect of oral administration of a Bacillus pumilus spore suspension on growth performance and health indicators. Treatments (n = 16) were: (1) non-medicated diet; (2) medicated diet with apramycin (200 mg/kg) and pharmacological levels of zinc oxide (2,500 mg zinc/kg) and (3) B. pumilus diet (non-medicated diet + 10(10) spores/day B. pumilus). Final body weight and average daily gain tended to be lower (P = 0.07) and feed conversion ratio was worsened (P<0.05) for the medicated treatment compared to the B. pumilus treatment. Ileal E. coli counts were lower for the B. pumilus and medicated treatments compared to the non-medicated treatment (P<0.05), perhaps as a result of increased ileal propionic acid concentrations (P<0.001). However, the medicated treatment reduced fecal (P<0.001) and cecal (P<0.05) Lactobacillus counts and tended to reduce the total cecal short chain fatty acid (SCFA) concentration (P = 0.10). Liver weights were lighter and concentrations of liver enzymes higher (P<0.05) in pigs on the medicated treatment compared to those on the non-medicated or B. pumilus treatments. Pigs on the B. pumilus treatment had lower overall lymphocyte and higher granulocyte percentages (P<0.001) and higher numbers of jejunal goblet cells (P<0.01) than pigs on either of the other two treatments or the non-medicated treatment, respectively. However, histopathological examination of the small intestine, kidneys and liver revealed no abnormalities. Overall, the B. pumilus treatment decreased ileal E. coli counts in a manner similar to the medicated treatment but without the adverse effects on growth performance, Lactobacillus counts, cecal SCFA concentration and possible liver toxicity experienced with the medicated treatment. PMID:24586349

  7. Diversity of Endophytic Fungi Isolated from Korean Ginseng Leaves

    PubMed Central

    Eo, Ju-Kyeong; Choi, Min-Seok

    2014-01-01

    We investigated the diversity of the foliar endophytes of Korean ginseng. Endophytic fungi were isolated from healthy leaves of mountain-cultivated ginseng (MCG) and field-cultivated ginseng (FCG) at 4 sites in Chungbuk Province. A total of 24 species of fungal endophytes were identified using molecular approaches. Additionally, the diversity of these endophytic fungi was compared between MCG and FCG. The major isolated endophytes were Edenia gomezpompae and Gibberella moniliformis in the MCG and FCG samples, respectively. The results suggest that ginseng endophytes have different community structures in different environments, and this understanding may prove useful in ginseng cultivation. PMID:25071383

  8. Antifungal and antibacterial metabolites from an endophytic Aspergillus sp. associated with Melia azedarach.

    PubMed

    Xiao, Jian; Zhang, Qiang; Gao, Yu-Qi; Shi, Xin-Wei; Gao, Jin-Ming

    2014-01-01

    Seven known metabolites, dianhydro-aurasperone C (1), isoaurasperone A (2), fonsecinone A (3), asperpyrone A (4), asperazine (5), rubrofusarin B (6) and (R)-3-hydroxybutanonitrile (7), were isolated from the culture of Aspergillus sp. KJ-9, a fungal endophyte isolated from Melia azedarach and identified by spectroscopic methods. All isolates were evaluated in vitro against several phytopathogenic fungi (Gibberella saubinetti, Magnaporthe grisea, Botrytis cinerea, Colletotrichum gloeosporioides and Alternaria solani) and pathogenic bacteria (Escherichia coli, Bacillus subtilis, Staphyloccocus aureus and Bacillus cereus). Compounds 3 and 7 were active against almost all phytopathogenic fungi tested with minimum inhibitory concentration (MIC) range of 6.25-50 ?M. Moreover, compound 3 was active against all pathogenic bacteria with MIC in the range of 25-100 ?M. Compound 7 is a rare new natural product isolated from a natural source for the first time, and the detailed NMR data of 1 were first assigned. PMID:24708541

  9. Endophytic bacteria from wheat grain as biocontrol agents of Fusarium graminearum and deoxynivalenol production in wheat.

    PubMed

    Pan, D; Mionetto, A; Tiscornia, S; Bettucci, L

    2015-08-01

    In Uruguay, Fusarium graminearum is the most common species that infects wheat and is responsible for Fusarium head blight (FHB) and contamination of grain with deoxynivalenol (DON). The aim of this work was to select bacterial endophytes isolated from wheat grain to evaluate their antagonistic ability against F. graminearum and DON production in vitro and under field conditions. Four strains identified as Bacillus megaterium (BM1) and Bacillus subtilis (BS43, BSM0 y BSM2) significantly reduced fungal growth and spore germination of F. graminearum. This antagonist activity remained unchanged after the bacterial cultures were heat treated. Under field conditions, treatments with antagonist BM1 was the most effective, reducing the FHB incidence and severity by 93 and 54 %, respectively, and the production of DON by 89.3 %. PMID:25956808

  10. Isolation, Characterization, and Insecticidal Activity of an Endophyte of Drunken Horse Grass, Achnatherum inebrians

    PubMed Central

    Shi, YingWu; Zhang, Xuebing; Lou, Kai

    2013-01-01

    Endophytic microorganisms reside within plant tissues and have often been found to promote plant growth. In this study, endophytic microorganisms were isolated from the roots, stems, leaves, and seeds of healthy drunken horse grass, Achnatherum inebrians (Hance) Keng (Poales: Poaceae), through the use of a grinding separation method and identified by a dual approach of morphological and physiological observation and 16S rRNA gene-based (for bacteria) and internal transcribed sequence-based (for fungi) molecular identification. The endophytes were then inoculated into liquid media for fermentation, and their crude extracts were employed for insecticidal activity tests using slide disc immersion and nebulization methods. A total of 89 bacteria species, which were classified into eight genera, Bacillus, Pseudomonas, Actinomyces, Corynebacterium, Acinetobacter, Sphingomonas, Paenibacillus, and Phyllobacterium, and two fungi, Claviceps and Chaetomium, were isolated. Of these species, isolates Streptomyces albus (Rossi-Doria) Waksman and Henrici (Actinomycetales: Streptomycetaceae) (GA) and Claviceps purpurea (Fr.) Tul. (Hypocreales: Clavicipitaceae) (PF-2) were shown to produce mortality rates of more than 90% in the cotton aphid, Aphis gossypii Glover (Hemiptera: Aphididae), after first and second screenings. The isolates PF-2 and GA associated with A. inebrians had significant insecticidal activities towards A. gossypii Glover (Hemiptera: Aphididae) and may provide a new biological resource for exploring a new microbial insecticide. PMID:24784492

  11. Metagenomics study of endophytic bacteria in Aloe vera using next-generation technology

    PubMed Central

    Akinsanya, Mushafau Adewale; Goh, Joo Kheng; Lim, Siew Ping; Ting, Adeline Su Yien

    2015-01-01

    Next generation sequencing (NGS) enables rapid analysis of the composition and diversity of microbial communities in several habitats. We applied the high throughput techniques of NGS to the metagenomics study of endophytic bacteria in Aloe vera plant, by assessing its PCR amplicon of 16S rDNA sequences (V3–V4 regions) with the Illumina metagenomics technique used to generate a total of 5,199,102 reads from the samples. The analyses revealed Proteobacteria, Firmicutes, Actinobacteria and Bacteriodetes as the predominant genera. The roots have the largest composition with 23% not present in other tissues. The stems have more of the genus—Pseudomonas and the unclassified Pseudomonadaceae. The α-diversity analysis indicated the richness and inverse Simpson diversity index of the bacterial endophyte communities for the leaf, root and stem tissues to be 2.221, 6.603 and 1.491 respectively. In a similar study on culturable endophytic bacteria in the same A. vera plants (unpublished work), the dominance of Pseudomonas and Bacillus genera was similar, with equal proportion of four species each in root, stem and leaf tissues. It is evident that NGS technology captured effectively the metagenomics of microbiota in plant tissues and this can improve our understanding of the microbial–plant host interactions. PMID:26697361

  12. Metagenomics study of endophytic bacteria in Aloe vera using next-generation technology.

    PubMed

    Akinsanya, Mushafau Adewale; Goh, Joo Kheng; Lim, Siew Ping; Ting, Adeline Su Yien

    2015-12-01

    Next generation sequencing (NGS) enables rapid analysis of the composition and diversity of microbial communities in several habitats. We applied the high throughput techniques of NGS to the metagenomics study of endophytic bacteria in Aloe vera plant, by assessing its PCR amplicon of 16S rDNA sequences (V3-V4 regions) with the Illumina metagenomics technique used to generate a total of 5,199,102 reads from the samples. The analyses revealed Proteobacteria, Firmicutes, Actinobacteria and Bacteriodetes as the predominant genera. The roots have the largest composition with 23% not present in other tissues. The stems have more of the genus-Pseudomonas and the unclassified Pseudomonadaceae. The α-diversity analysis indicated the richness and inverse Simpson diversity index of the bacterial endophyte communities for the leaf, root and stem tissues to be 2.221, 6.603 and 1.491 respectively. In a similar study on culturable endophytic bacteria in the same A. vera plants (unpublished work), the dominance of Pseudomonas and Bacillus genera was similar, with equal proportion of four species each in root, stem and leaf tissues. It is evident that NGS technology captured effectively the metagenomics of microbiota in plant tissues and this can improve our understanding of the microbial-plant host interactions. PMID:26697361

  13. Antitumor and antimicrobial activities of endophytic fungi from medicinal parts of Aquilaria sinensis.

    PubMed

    Cui, Jin-long; Guo, Shun-xing; Xiao, Pei-gen

    2011-05-01

    The purpose of this study was to isolate and characterize endophytic fungi from the stem tissue which can produce fragrant ingredients in Aquilaria sinensis (also called agarwood) to determine their antitumor and antimicrobial activities. Twenty-eight fungal endophytes were isolated from agarwood by strict sterile sample preparation and were classified into 14 genera and 4 taxonomic classes (Sordariomycetes, Dothideomycetes, Saccharomycetes, and Zygomycetes) based on molecular identification. Of the 28 isolates, 13 (46.4%) showed antimicrobial activity against at least one of the test strains by the agar well diffusion method, and 23 isolates (82.1%) displayed antitumor activity against at least one of five cancer cell lines by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The diameters of inhibition zones of YNAS07, YNAS14, HNAS04, HNAS05, HNAS08, and HNAS11 were equal to or higher than 14.0 mm against Staphylococcus aureus, Escherichia coli, Bacillus subtilis, B. subtilis, Aspergillus fumigatus, and B. subtilis, respectively. The inhibition rates of YNAS06, YNAS08, and HNAS06 were not less than 60% to 293-T, 293-T, and SKVO3 cells, respectively. These results suggest that the endophytic fungi associated with agarwood will provide us with not only useful micro-ecological information, but also potential antimicrobial and antitumor agents. PMID:21528493

  14. Antitumor and antimicrobial activities of endophytic fungi from medicinal parts of Aquilaria sinensis *

    PubMed Central

    Cui, Jin-long; Guo, Shun-xing; Xiao, Pei-gen

    2011-01-01

    The purpose of this study was to isolate and characterize endophytic fungi from the stem tissue which can produce fragrant ingredients in Aquilaria sinensis (also called agarwood) to determine their antitumor and antimicrobial activities. Twenty-eight fungal endophytes were isolated from agarwood by strict sterile sample preparation and were classified into 14 genera and 4 taxonomic classes (Sordariomycetes, Dothideomycetes, Saccharomycetes, and Zygomycetes) based on molecular identification. Of the 28 isolates, 13 (46.4%) showed antimicrobial activity against at least one of the test strains by the agar well diffusion method, and 23 isolates (82.1%) displayed antitumor activity against at least one of five cancer cell lines by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The diameters of inhibition zones of YNAS07, YNAS14, HNAS04, HNAS05, HNAS08, and HNAS11 were equal to or higher than 14.0 mm against Staphylococcus aureus, Escherichia coli, Bacillus subtilis, B. subtilis, Aspergillus fumigatus, and B. subtilis, respectively. The inhibition rates of YNAS06, YNAS08, and HNAS06 were not less than 60% to 293-T, 293-T, and SKVO3 cells, respectively. These results suggest that the endophytic fungi associated with agarwood will provide us with not only useful micro-ecological information, but also potential antimicrobial and antitumor agents. PMID:21528493

  15. Identification and characterization of endophytic bacteria from corn (Zea mays L.) roots with biotechnological potential in agriculture

    PubMed Central

    2014-01-01

    Six endophytic bacteria of corn roots were identified as Bacillus sp. and as Enterobacter sp, by sequencing of the 16S rRNA gene. Four of the strains, CNPSo 2476, CNPSo 2477, CNPSo 2478 and CNPSo 2480 were positive for the nitrogen fixation ability evaluated through the acetylene reduction assay and amplification of nifH gene. Two Bacillus strains (CNPSo 2477 and CNPSo 2478) showed outstanding skills for the production of IAA, siderophores and lytic enzymes, but were not good candidates as growth promoters, because they reduced seed germination. However, the same strains were antagonists against the pathogenic fungi Fusarium verticillioides, Colletotrichum graminicola, Bipolaris maydis and Cercospora zea-maydis. As an indication of favorable bacterial action, Enterobacter sp. CNPSo 2480 and Bacillus sp. CNPSo 2481 increased the root volume by 44% and 39%, respectively, and the seed germination by 47% and 56%, respectively. Therefore, these two strains are good candidates for future testing as biological inoculants for corn. PMID:24949261

  16. Enzymes of the Tryptophan Pathway in Three Bacillus Species

    PubMed Central

    Hoch, Sallie O'Neil; Crawford, Irving P.

    1973-01-01

    The tryptophan synthetic pathway was characterized in three species of Bacillus, B. subtilis, B. pumilus, and B. alvei. They share the common features of a pathway which is subject to tryptophan repression, contains no unexpected complexes among the five enzymes, exhibits dissociable anthranilate synthase enzymes which do not require phosphoribosyl transferase for amidetransfer activity, contains separate indoleglycerol phosphate synthase and phosphoribosylanthranilate isomerase enzymes, and contains similar tryptophan synthetase multimers. In looking at these characteristics in detail however, differences among the three species became apparent, as, for example, in the complementation observed between the ? and ?2 components of tryptophan synthetase, and the dissociation patterns of the large and small components of anthranilate synthase. The results demonstrate some pitfalls in attempting to compare multimeric enzymes in crude extracts from different organisms. PMID:4200852

  17. A PCR test to identify bacillus subtilis and closely related species and its application to the monitoring of wastewater biotreatment.

    PubMed

    Wattiau, P; Renard, M E; Ledent, P; Debois, V; Blackman, G; Agathos, S N

    2001-09-01

    A PCR test based on the 16S rRNA gene was set up that could identify any of the five species of the 'Bacillus subtilis group' (B. subtilis, B. pumilus, B. atrophaeus, B. lichenijormis and B. amyloliquefaciens). The test was directly applicable to single colonies and showed excellent specificity. In the mixed population context of wastewater analysis, direct detection of the target Bacillus species by PCR on either crude or purified DNA extracts had poor sensitivity. When assayed on cell suspensions derived from enriched wastewater samples, sensitivity was increased. Using a simple calibration method, it was possible to estimate the proportion of the target organisms. This method was found suitable for easy monitoring of a wastewater bioaugmentation experiment carried out with a mixture of sporulated Bacillus strains. PMID:11601635

  18. Endophytic bacteria take the challenge to improve Cu phytoextraction by sunflower.

    PubMed

    Kolbas, Aliaksandr; Kidd, Petra; Guinberteau, Jacques; Jaunatre, Renaud; Herzig, Rolf; Mench, Michel

    2015-04-01

    Endophytic bacteria from roots and crude seed extracts of a Cu-tolerant population of Agrostis capillaris were inoculated to a sunflower metal-tolerant mutant line, and their influence on Cu tolerance and phytoextraction was assessed using a Cu-contaminated soil series. Ten endophytic bacterial strains isolated from surface-sterilized A. capillaris roots were mixed to prepare the root endophyte inoculant (RE). In parallel, surface-sterilized seeds of A. capillaris were crushed in MgSO4 to prepare a crude seed extract containing seed endophytes (SE). An aliquot of this seed extract was filtered at 0.2 ?m to obtain a bacterial cell-free seed extract (SEF). After surface sterilization, germinated sunflower seeds were separately treated with one of five modalities: no treatment (C), immersion in MgSO4 (CMg) or SEF solutions and inoculation with RE or SE. All plants were cultivated on a Cu-contaminated soil series (13-1020 mg Cu kg(-1)). Cultivable RE strains were mostly members of the Pseudomonas genera, and one strain was closely related to Labrys sp. The cultivable SE strains belonged mainly to the Bacillus genera and some members of the Rhodococcus genera. The treatment effects depended on the soil Cu concentration. Both SE and SEF plants had a higher Cu tolerance in the 13-517 mg Cu kg(-1) soil range as reflected by increased shoot and root DW yields compared to control plants. This was accompanied by a slight decrease in shoot Cu concentration and increase in root Cu concentration. Shoot and root DW yields were more promoted by SE than SEF in the 13-114 mg Cu kg(-1) soil range, which could reflect the influence of seed-located bacterial endophytes. At intermediate soil Cu (416-818 mg Cu kg(-1) soil), the RE and CMg plants had lower shoot Cu concentrations than the control, SE and SEF plants. At high total soil Cu (617-1020 mg Cu kg(-1)), root DW yield of RE plants slightly increased and their root Cu concentration rose by up to 1.9-fold. In terms of phytoextraction efficiency, shoot Cu removal was increased for sunflower plants inoculated with crude and bacterial cell-free seed extracts by 1.3- to 2.2-fold in the 13-416 mg Cu kg(-1) soil range. Such increase was mainly driven by an enhanced shoot DW yield. The number and distribution of endophytic bacteria in the harvested sunflower tissues must be further examined. PMID:25561255

  19. Enterotoxins and emetic toxins production by Bacillus cereus and other species of Bacillus isolated from Soumbala and Bikalga, African alkaline fermented food condiments.

    PubMed

    Ouoba, Labia Irene I; Thorsen, Line; Varnam, Alan H

    2008-06-10

    The ability of various species of Bacillus from fermented seeds of Parkia biglobosa known as African locust bean (Soumbala) and fermented seeds of Hibiscus sabdariffa (Bikalga) was investigated. The study included screening of the isolates by haemolysis on blood agar, detection of toxins in broth and during the fermentation of African locust bean using the Bacillus cereus Enterotoxin Reverse Passive Latex Agglutination test kit (BCET-RPLA) and the Bacillus Diarrhoeal Enterotoxin Visual Immunoassay (BDEVIA). Detection of genes encoding cytotoxin K (CytK), haemolysin BL (Hbl A, Hbl C, Hbl D), non-hemolytic enterotoxin (NheA, NheB, NheC) and EM1 specific of emetic toxin producers was also investigated using PCR with single pair and multiplex primers. Of 41 isolates, 29 Bacillus belonging to the species of B. cereus, Bacillus subtilis, Bacillus licheniformis and Bacillus pumilus showed haemolysis on blood agar. Using RPLA, enterotoxin production was detected for three isolates of B. cereus in broth and all B. cereus (9) in fermented seeds. Using BDEVIA, enterotoxin production was detected in broth as well as in fermented seeds for all B. cereus isolates. None of the isolates belonging to the other Bacillus species was able to produce enterotoxins either by RPLA or BDEVIA. Nhe genes were detected in all B. cereus while Hbl and CytK genes were detected respectively in five and six B. cereus strains. A weak presence of Hbl (A, D) and CytK genes was detected in two isolates of B. subtilis and one of B. licheniformis but results were inconsistent, especially for Hbl genes. The emetic specific gene fragment EM1 was not detected in any of the isolates studied. PMID:18474404

  20. Genomics of Bacillus Species

    NASA Astrophysics Data System (ADS)

    Økstad, Ole Andreas; Kolstø, Anne-Brit

    Members of the genus Bacillus are rod-shaped spore-forming bacteria belonging to the Firmicutes, the low G+C gram-positive bacteria. The Bacillus genus was first described and classified by Ferdinand Cohn in Cohn (1872), and Bacillus subtilis was defined as the type species (Soule, 1932). Several Bacilli may be linked to opportunistic infections. However, pathogenicity among Bacillus spp. is mainly a feature of bacteria belonging to the Bacillus cereus group, including B. cereus, Bacillus anthracis, and Bacillus thuringiensis. Here we review the genomics of B. cereus group bacteria in relation to their roles as etiological agents of two food poisoning syndromes (emetic and diarrhoeal).

  1. Phylogeny in Aid of the Present and Novel Microbial Lineages: Diversity in Bacillus

    PubMed Central

    Porwal, Shalini; Lal, Sadhana; Cheema, Simrita; Kalia, Vipin Chandra

    2009-01-01

    Bacillus represents microbes of high economic, medical and biodefense importance. Bacillus strain identification based on 16S rRNA sequence analyses is invariably limited to species level. Secondly, certain discrepancies exist in the segregation of Bacillus subtilis strains. In the RDP/NCBI databases, out of a total of 2611 individual 16S rDNA sequences belonging to the 175 different species of the genus Bacillus, only 1586 have been identified up to species level. 16S rRNA sequences of Bacillus anthracis (153 strains), B. cereus (211 strains), B. thuringiensis (108 strains), B. subtilis (271 strains), B. licheniformis (131 strains), B. pumilus (83 strains), B. megaterium (47 strains), B. sphaericus (42 strains), B. clausii (39 strains) and B. halodurans (36 strains) were considered for generating species-specific framework and probes as tools for their rapid identification. Phylogenetic segregation of 1121, 16S rDNA sequences of 10 different Bacillus species in to 89 clusters enabled us to develop a phylogenetic frame work of 34 representative sequences. Using this phylogenetic framework, 305 out of 1025, 16S rDNA sequences presently classified as Bacillus sp. could be identified up to species level. This identification was supported by 20 to 30 nucleotides long signature sequences and in silico restriction enzyme analysis specific to the 10 Bacillus species. This integrated approach resulted in identifying around 30% of Bacillus sp. up to species level and revealed that B. subtilis strains can be segregated into two phylogenetically distinct groups, such that one of them may be renamed. PMID:19212464

  2. Phylogeny in aid of the present and novel microbial lineages: diversity in Bacillus.

    PubMed

    Porwal, Shalini; Lal, Sadhana; Cheema, Simrita; Kalia, Vipin Chandra

    2009-01-01

    Bacillus represents microbes of high economic, medical and biodefense importance. Bacillus strain identification based on 16S rRNA sequence analyses is invariably limited to species level. Secondly, certain discrepancies exist in the segregation of Bacillus subtilis strains. In the RDP/NCBI databases, out of a total of 2611 individual 16S rDNA sequences belonging to the 175 different species of the genus Bacillus, only 1586 have been identified up to species level. 16S rRNA sequences of Bacillus anthracis (153 strains), B. cereus (211 strains), B. thuringiensis (108 strains), B. subtilis (271 strains), B. licheniformis (131 strains), B. pumilus (83 strains), B. megaterium (47 strains), B. sphaericus (42 strains), B. clausii (39 strains) and B. halodurans (36 strains) were considered for generating species-specific framework and probes as tools for their rapid identification. Phylogenetic segregation of 1121, 16S rDNA sequences of 10 different Bacillus species in to 89 clusters enabled us to develop a phylogenetic frame work of 34 representative sequences. Using this phylogenetic framework, 305 out of 1025, 16S rDNA sequences presently classified as Bacillus sp. could be identified up to species level. This identification was supported by 20 to 30 nucleotides long signature sequences and in silico restriction enzyme analysis specific to the 10 Bacillus species. This integrated approach resulted in identifying around 30% of Bacillus sp. up to species level and revealed that B. subtilis strains can be segregated into two phylogenetically distinct groups, such that one of them may be renamed. PMID:19212464

  3. Expressed nifH Genes of Endophytic Bacteria Detected in Field-Grown Sweet Potatoes (Ipomoea batatas L.).

    PubMed

    Terakado-Tonooka, Junko; Ohwaki, Yoshinari; Yamakawa, Hiromoto; Tanaka, Fukuyo; Yoneyama, Tadakatsu; Fujihara, Shinsuke

    2008-01-01

    We examined the nitrogenase reductase (nifH) genes of endophytic diazotrophic bacteria expressed in field-grown sweet potatoes (Ipomoea batatas L.) by reverse transcription (RT)-PCR. Gene fragments corresponding to nifH were amplified from mRNA obtained from the stems and storage roots of field-grown sweet potatoes several months after planting. Sequence analysis revealed that these clones were homologous to the nifH sequences of Bradyrhizobium, Pelomonas, and Bacillus sp. in the DNA database. Investigation of the nifH genes amplified from the genomic DNA extracted from these sweet potatoes also showed high similarity to various ?-proteobacteria including Bradyrhizobium, ?-proteobacteria, and cyanobacteria. These results suggest that bradyrhizobia colonize and express nifH genes not only in the root nodules of leguminous plants but also in sweet potatoes as diazotrophic endophytes. PMID:21558693

  4. Antagonism of entomopathogenic fungi by Bacillus spp. associated with the integument of cicadellids and delphacids.

    PubMed

    Toledo, Andrea; López, Silvina; Aulicino, Mónica; de Remes-Lenicov, Ana María; Balatti, Pedro

    2015-06-01

    Entomopathogenic fungi are potential tools to biocontrol cicadellids and delphacids, two groups of insects that cause extensive damage to agricultural crops. However, bacteria living on the host cuticle may inhibit fungal growth. In the present work, following the molecular characterization of 10 strains of Bacillus isolated from the integument of cicadellids and delphacids, we selected isolates of the fungi Beauveria bassiana and Metarhizium anisopliae that are resistant to the antimicrobials secreted by these bacterial strains. The antagonistic activity of the 10 bacterial isolates belonging to the genus Bacillus (i.e., B. amyloliquefaciens, B. pumilus, and B. subtilis) against 41 isolates of Bea. bassiana and 20 isolates of M. anisopliae was investigated in vitro on tryptic soy agar using the central disk test. With this approach, isolates of Bea. bassiana and M. anisopliae resistant to antagonistic bacteria were identified that can be further developed as biological control agents. PMID:26496616

  5. Fungal endophytes limit pathogen damage in a tropical tree

    PubMed Central

    Arnold, A. Elizabeth; Mejía, Luis Carlos; Kyllo, Damond; Rojas, Enith I.; Maynard, Zuleyka; Robbins, Nancy; Herre, Edward Allen

    2003-01-01

    Every plant species examined to date harbors endophytic fungi within its asymptomatic aerial tissues, such that endophytes represent a ubiquitous, yet cryptic, component of terrestrial plant communities. Fungal endophytes associated with leaves of woody angiosperms are especially diverse; yet, fundamental aspects of their interactions with hosts are unknown. In contrast to the relatively species-poor endophytes that are vertically transmitted and act as defensive mutualists of some temperate grasses, the diverse, horizontally transmitted endophytes of woody angiosperms are thought to contribute little to host defense. Here, we document high diversity, spatial structure, and host affinity among foliar endophytes associated with a tropical tree (Theobroma cacao, Malvaceae) across lowland Panama. We then show that inoculation of endophyte-free leaves with endophytes isolated frequently from naturally infected, asymptomatic hosts significantly decreases both leaf necrosis and leaf mortality when T. cacao seedlings are challenged with a major pathogen (Phytophthora sp.). In contrast to reports of fungal inoculation inducing systemic defense, we found that protection was primarily localized to endophyte-infected tissues. Further, endophyte-mediated protection was greater in mature leaves, which bear less intrinsic defense against fungal pathogens than do young leaves. In vitro studies suggest that host affinity is mediated by leaf chemistry, and that protection may be mediated by direct interactions of endophytes with foliar pathogens. Together, these data demonstrate the capacity of diverse, horizontally transmitted endophytes of woody angiosperms to play an important but previously unappreciated role in host defense. PMID:14671327

  6. Analysis of the abilities of endophytic bacteria associated with banana tree roots to promote plant growth.

    PubMed

    Andrade, Leandro Fernandes; de Souza, Gleika Larisse Oliveira Dorasio; Nietsche, Silvia; Xavier, Adelica Aparecida; Costa, Marcia Regina; Cardoso, Acleide Maria Santos; Pereira, Marlon Cristian Toledo; Pereira, Débora Francine Gomes Silva

    2014-01-01

    A total of 40 endophytic bacterial isolates obtained from banana tree roots were characterized for their biotechnological potential for promoting banana tree growth. All isolates had at least one positive feature. Twenty isolates were likely diazotrophs and formed pellicles in nitrogen-free culture medium, and 67% of these isolates belonged to the genus Bacillus sp. The isolates EB-04, EB-169, EB-64, and EB-144 had N fixation abilities as measured by the Kjeldahl method and by an acetylene reduction activity assay. Among the 40 isolates, 37.5% were capable of solubilizing inorganic phosphate and the isolates EB-47 and EB-64 showed the highest solubilization capacity. The isolate EB-53 (Lysinibacillus sp.) had a high solubilization index, whereas 73% of the isolates had low solubilization indices. The synthesis of indole-3-acetic acid (IAA) in the presence of L-tryptophan was detected in 40% of the isolates. The isolate EB-40 (Bacillus sp.) produced the highest amount of IAA (47.88 ?g/ml) in medium supplemented with L-tryptophan and was able to synthesize IAA in the absence of L-tryptophan. The isolates EB-126 (Bacillus subtilis) and EB-47 (Bacillus sp.) were able to simultaneously fix nitrogen, solubilize phosphate and produce IAA in vitro. The results of this study demonstrated that the isolates analyzed here had diverse abilities and all have the potential to be used as growth-promoting microbial inoculants for banana trees. PMID:24390835

  7. Bacillus subtilis

    PubMed Central

    Amuguni, Hellen; Tzipori, Saul

    2012-01-01

    Most pathogens enter the body through mucosal surfaces. Mucosal immunization, a non-invasive needle-free route, often stimulates a mucosal immune response that is both effective against mucosal and systemic pathogens. The development of mucosally administered heat-stable vaccines with long shelf life would therefore significantly enhance immunization programs in developing countries by avoiding the need for a cold chain or systemic injections. Currently, recombinant vaccine carriers are being used for antigen delivery. Engineering Bacillus subtilis for use as a non-invasive and heat stable antigen delivery system has proven successful. Bacterial spores protected by multiple layers of protein are known to be robust and resistant to desiccation. Stable constructs have been created by integration into the bacterial chromosome of immunogens. The spore coat has been used as a vehicle for heterologous antigen presentation and protective immunization. Sublingual (SL) and intranasal (IN) routes have recently received attention as delivery routes for therapeutic drugs and vaccines and recent attempts by several investigators, including our group, to develop vaccines that can be delivered intranasally and sublingually have met with a lot of success. As discussed in this Review, the use of Bacillus subtilis to express antigens that can be administered either intranasally or sublingually is providing new insights in the area of mucosal vaccines. In our work, we evaluated the efficacy of SL and IN immunizations with B. subtilis engineered to express tetanus toxin fragment C (TTFC) in mice and piglets. These bacteria engineered to express heterologous antigen either on the spore surface or within the vegetative cell have been used for oral, IN and SL delivery of antigens. A Bacillus subtilis spore coat protein, CotC was used as a fusion partner to express the tetanus fragment C. B. subtilis spores known to be highly stable and safe are also easy to purify making this spore-based display system a potentially powerful approach for surface expression of antigens. These advances will help to accelerate the development and testing of new mucosal vaccines against many human and animal diseases. PMID:22699442

  8. Bacillus xiamenensis sp. nov., isolated from intestinal tract contents of a flathead mullet (Mugil cephalus).

    PubMed

    Lai, Qiliang; Liu, Yang; Shao, Zongze

    2014-01-01

    A taxonomic study was carried out on strain HYC-10(T), which was isolated from the intestinal tract contents of a flathead mullet, Mugil cephalus, captured from the sea off Xiamen Island, China. The bacterium was observed to be Gram positive, oxidase and catalase positive, rod shaped, and motile by subpolar flagella. The bacterium was found to grow at salinities of 0-12 % and at temperatures of 8-45 °C. The isolate was found to hydrolyze aesculin and gelatin, but was unable to reduce nitrate to nitrite. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain HYC-10(T) belongs to the genus Bacillus, with highest sequence similarity (99.3 %) to Bacillus aerophilus 28K(T), Bacillus stratosphericus 41KF2a(T) and Bacillus altitudinis DSM 21631(T), followed by Bacillus safensis DSM 19292(T) (99.5 %) and Bacillus pumilus DSM 27(T) (99.5 %), while the sequence similarities to others were all below 97.6 %. The genomic ANIm values between strain HYC-10(T) and three type strains (B. altitudinis DSM 21631(T), B. safensis DSM 19292(T) and B. pumilus DSM 27(T)) were determined to range from 89.11 to 91.53 %. The DNA-DNA hybridization estimate values between strain HYC-10(T) and the three type strains were from 36.60 to 44.00 %. The principal fatty acids identified were iso-C15:0 (39.1 %), anteiso-C15:0 (22.7 %), iso-C17:0 (13.1 %), C16:0 (6.1 %), anteiso-C17:0 (5.8 %) and iso-C16:0 (5.1 %). The G+C content of the chromosomal DNA was determined from the draft genome sequence to be 41.3 mol%. The respiratory quinone was determined to be MK-7 (100 %). Phosphatidylglycerol, diphosphatidylglycerol, aminoglycolipid, two glycolipids and two unknown phospholipids were found to be present. The combined genotypic and phenotypic data show that strain HYC-10(T) represents a novel species of the genus Bacillus, for which the name Bacillus xiamenensis sp. nov. is proposed, with the type strain HYC-10(T) (=CGMCC NO.1.12326(T) = LMG 27143(T) = MCCC 1A00008(T)). PMID:24158533

  9. Selenium hyperaccumulators harbor a diverse endophytic bacterial community characterized by high selenium resistance and plant growth promoting properties

    PubMed Central

    Sura-de Jong, Martina; Reynolds, Ray J. B.; Richterova, Klara; Musilova, Lucie; Staicu, Lucian C.; Chocholata, Iva; Cappa, Jennifer J.; Taghavi, Safiyh; van der Lelie, Daniel; Frantik, Tomas; Dolinova, Iva; Strejcek, Michal; Cochran, Alyssa T.; Lovecka, Petra; Pilon-Smits, Elizabeth A. H.

    2015-01-01

    Selenium (Se)-rich plants may be used to provide dietary Se to humans and livestock, and also to clean up Se-polluted soils or waters. This study focused on endophytic bacteria of plants that hyperaccumulate selenium (Se) to 0.5–1% of dry weight. Terminal restriction fragment length polymorphism (T-RFLP) analysis was used to compare the diversity of endophytic bacteria of hyperaccumulators Stanleya pinnata (Brassicaceae) and Astragalus bisulcatus (Fabaceae) with those from related non-accumulators Physaria bellii (Brassicaceae) and Medicago sativa (Fabaceae) collected on the same, seleniferous site. Hyperaccumulators and non-accumulators showed equal T-RF diversity. Parsimony analysis showed that T-RFs from individuals of the same species were more similar to each other than to those from other species, regardless of plant Se content or spatial proximity. Cultivable endophytes from hyperaccumulators S. pinnata and A. bisulcatus were further identified and characterized. The 66 bacterial morphotypes were shown by MS MALDI-TOF Biotyper analysis and 16S rRNA gene sequencing to include strains of Bacillus, Pseudomonas, Pantoea, Staphylococcus, Paenibacillus, Advenella, Arthrobacter, and Variovorax. Most isolates were highly resistant to selenate and selenite (up to 200 mM) and all could reduce selenite to red elemental Se, reduce nitrite and produce siderophores. Seven isolates were selected for plant inoculation and found to have plant growth promoting properties, both in pure culture and when co-cultivated with crop species Brassica juncea (Brassicaceae) or M. sativa. There were no effects on plant Se accumulation. We conclude that Se hyperaccumulators harbor an endophytic bacterial community in their natural seleniferous habitat that is equally diverse to that of comparable non-accumulators. The hyperaccumulator endophytes are characterized by high Se resistance, capacity to produce elemental Se and plant growth promoting properties. PMID:25784919

  10. Selenium hyperaccumulators harbor a diverse endophytic bacterial community characterized by high selenium resistance and plant growth promoting properties.

    PubMed

    Sura-de Jong, Martina; Reynolds, Ray J B; Richterova, Klara; Musilova, Lucie; Staicu, Lucian C; Chocholata, Iva; Cappa, Jennifer J; Taghavi, Safiyh; van der Lelie, Daniel; Frantik, Tomas; Dolinova, Iva; Strejcek, Michal; Cochran, Alyssa T; Lovecka, Petra; Pilon-Smits, Elizabeth A H

    2015-01-01

    Selenium (Se)-rich plants may be used to provide dietary Se to humans and livestock, and also to clean up Se-polluted soils or waters. This study focused on endophytic bacteria of plants that hyperaccumulate selenium (Se) to 0.5-1% of dry weight. Terminal restriction fragment length polymorphism (T-RFLP) analysis was used to compare the diversity of endophytic bacteria of hyperaccumulators Stanleya pinnata (Brassicaceae) and Astragalus bisulcatus (Fabaceae) with those from related non-accumulators Physaria bellii (Brassicaceae) and Medicago sativa (Fabaceae) collected on the same, seleniferous site. Hyperaccumulators and non-accumulators showed equal T-RF diversity. Parsimony analysis showed that T-RFs from individuals of the same species were more similar to each other than to those from other species, regardless of plant Se content or spatial proximity. Cultivable endophytes from hyperaccumulators S. pinnata and A. bisulcatus were further identified and characterized. The 66 bacterial morphotypes were shown by MS MALDI-TOF Biotyper analysis and 16S rRNA gene sequencing to include strains of Bacillus, Pseudomonas, Pantoea, Staphylococcus, Paenibacillus, Advenella, Arthrobacter, and Variovorax. Most isolates were highly resistant to selenate and selenite (up to 200 mM) and all could reduce selenite to red elemental Se, reduce nitrite and produce siderophores. Seven isolates were selected for plant inoculation and found to have plant growth promoting properties, both in pure culture and when co-cultivated with crop species Brassica juncea (Brassicaceae) or M. sativa. There were no effects on plant Se accumulation. We conclude that Se hyperaccumulators harbor an endophytic bacterial community in their natural seleniferous habitat that is equally diverse to that of comparable non-accumulators. The hyperaccumulator endophytes are characterized by high Se resistance, capacity to produce elemental Se and plant growth promoting properties. PMID:25784919

  11. Are Epichloë endophytes specific to Elymus grass hosts?

    PubMed

    Song, H; Song, Q Y; Li, X Z; Nan, Z B

    2015-01-01

    Epichloë endophytes are widely distributed mutualists of cool-season grasses and can protect their hosts against biotic and abiotic stresses. Previous studies have shown that Epichloë endophytes are specific to their grass hosts in tall Festuca and Lolium species. However, no systematic analysis exists of host specificity of asexual Epichloë endophytes and Chinese Elymus species. We analyzed the phylogenetic relationships between Chinese Elymus species and their diploid donor Hordeum species, using their corresponding Epichloë endophyte sequences. We found that 1) the maternal donor of the Chinese Elymus species was the Chinese Pseudoroegneria (St genome) or Hordeum (H genome); and 2) Chinese Hordeum species probably contained two species of Epichloë endophytes. One Epichloë endophyte was also present in a North American Elymus species. The other Epichloë endophyte was found in a Chinese Elymus species. Our results indicate that Epichloë endophytes isolated from Elymus species did not show grass-host specificity. 3) Plant hybridization could probably transform endophyte-free plants (E-) to endophyte-infected plants (E+). Based on our data, we formulate hypotheses about which Epichloë endophytes were spread via plant hybridization. PMID:26782389

  12. Endophytic fungi: resource for gibberellins and crop abiotic stress resistance.

    PubMed

    Khan, Abdul Latif; Hussain, Javid; Al-Harrasi, Ahmed; Al-Rawahi, Ahmed; Lee, In-Jung

    2015-03-01

    The beneficial effects of endophytes on plant growth are important for agricultural ecosystems because they reduce the need for fertilizers and decrease soil and water pollution while compensating for environmental perturbations. Endophytic fungi are a novel source of bioactive secondary metabolites; moreover, recently they have been found to produce physiologically active gibberellins as well. The symbiosis of gibberellins producing endophytic fungi with crops can be a promising strategy to overcome the adverse effects of abiotic stresses. The association of such endophytes has not only increased plant biomass but also ameliorated plant-growth during extreme environmental conditions. Endophytic fungi represent a trove of unexplored biodiversity and a frequently overlooked component of crop ecology. The present review describes the role of gibberellins producing endophytic fungi, suggests putative mechanisms involved in plant endophyte stress interactions and discusses future prospects in this field. PMID:23984800

  13. Bioactive metabolites from the endophytic fungus Alternaria alternata.

    PubMed

    Wang, Ying; Yang, Ming-Hua; Wang, Xiao-Bing; Li, Tian-Xiao; Kong, Ling-Yi

    2014-12-01

    Two altenuene derivatives (1-2) and one isocoumarin (3), together with six known compounds (4-9) were isolated from solid cultures of an endophytic fungus Alternaria alternata, obtained from the fresh branches of Camellia sinensis. Chiral analysis revealed the racemic nature of 1 and 2, which were subsequently resolved into two pairs of enantiomers [(+)-1 and (-)-1, (+)-2 and (-)-2]. Structures of all the isolates were identified through spectroscopic data. Absolute configurations of the two pairs of enantiomers were determined by electronic circular dichroism (ECD) calculation and the chiral center of C-10 in 3 was deduced via [Rh2(OCOCF?)?]-induced CD experiment. All the isolates were evaluated for their antimicrobial abilities against the pathogenic bacteria and fungi as well as cytotoxic activities against two human tumor cell lines. Compound 5 was the most active against Bacillus subtilis with MIC?? of 8.6 ?g/ml, and compounds 1-3, 6-7 and 9 exhibited moderate to weak inhibition towards the test pathogenic microorganism. Compound 4 showed mild cytotoxic activity against human osteosarcoma cells U2OS with IC?? of 28.3 ?M. PMID:25261763

  14. Bacillus thuringiensis

    PubMed Central

    Ibrahim, Mohamed A; Griko, Natalya; Junker, Matthew

    2010-01-01

    Bacillus thuringiensis (Bt) is a unique bacterium in that it shares a common place with a number of chemical compounds which are used commercially to control insects important to agriculture and public health. Although other bacteria, including B. popilliae and B. sphaericus, are used as microbial insecticides, their spectrum of insecticidal activity is quite limited compared to Bt. Importantly, Bt is safe for humans and is the most widely used environmentally compatible biopesticide worldwide. Furthermore, insecticidal Bt genes have been incorporated into several major crops, rendering them insect resistant, and thus providing a model for genetic engineering in agriculture. This review highlights what the authors consider the most relevant issues and topics pertaining to the genomics and proteomics of Bt. At least one of the authors (L.A.B.) has spent most of his professional life studying different aspects of this bacterium with the goal in mind of determining the mechanism(s) by which it kills insects. The other authors have a much shorter experience with Bt but their intellect and personal insight have greatly enriched our understanding of what makes Bt distinctive in the microbial world. Obviously, there is personal interest and bias reflected in this article notwithstanding oversight of a number of published studies. This review contains some material not published elsewhere although several ideas and concepts were developed from a broad base of scientific literature up to 2010. PMID:21327125

  15. Evolution of Reproductive Morphology in Leaf Endophytes

    PubMed Central

    Wang, Zheng; Johnston, Peter R.; Yang, Zhu L.; Townsend, Jeffrey P.

    2009-01-01

    The endophytic lifestyle has played an important role in the evolution of the morphology of reproductive structures (body) in one of the most problematic groups in fungal classification, the Leotiomycetes (Ascomycota). Mapping fungal morphologies to two groups in the Leiotiomycetes, the Rhytismatales and Hemiphacidiaceae reveals significant divergence in body size, shape and complexity. Mapping ecological roles to these taxa reveals that the groups include endophytic fungi living on leaves and saprobic fungi living on duff or dead wood. Finally, mapping of the morphologies to ecological roles reveals that leaf endophytes produce small, highly reduced fruiting bodies covered with fungal tissue or dead host tissue, while saprobic species produce large and intricate fruiting bodies. Intriguingly, resemblance between asexual conidiomata and sexual ascomata in some leotiomycetes implicates some common developmental pathways for sexual and asexual development in these fungi. PMID:19158947

  16. Metabolite and gene expression studies in endophyte infected and uninfected tall fescue under water deficit stress

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Tall fescue plants symbiotic with the endophytic fungus, Neotyphodium coenophialum (E+), have better survivability and persistence under stressful conditions, especially under drought stress, than plants lacking the endophyte (E-). To understand more about the grass-endophyte interactions, how endop...

  17. Diversity of endophytic fungi in Glycine max.

    PubMed

    Fernandes, Elio Gomes; Pereira, Olinto Liparini; Silva, Cynthia Cânedo da; Bento, Claudia Braga Pereira; Queiroz, Marisa Vieira de

    2015-12-01

    Endophytic fungi are microorganisms that live within plant tissues without causing disease during part of their life cycle. With the isolation and identification of these fungi, new species are being discovered, and ecological relationships with their hosts have also been studied. In Glycine max, limited studies have investigated the isolation and distribution of endophytic fungi throughout leaves and roots. The distribution of these fungi in various plant organs differs in diversity and abundance, even when analyzed using molecular techniques that can evaluate fungal communities in different parts of the plants, such as denaturing gradient gel electrophoresis (DGGE). Our results show there is greater species richness of culturable endophytic filamentous fungi in the leaves G. max as compared to roots. Additionally, the leaves had high values for diversity indices, i.e. Simpsons, Shannon and Equitability. Conversely, dominance index was higher in roots as compared to leaves. The fungi Ampelomyces sp., Cladosporium cladosporioides, Colletotrichum gloeosporioides, Diaporthe helianthi, Guignardia mangiferae and Phoma sp. were more frequently isolated from the leaves, whereas the fungi Fusarium oxysporum, Fusarium solani and Fusarium sp. were prevalent in the roots. However, by evaluating the two communities by DGGE, we concluded that the species richness was higher in the roots than in the leaves. UPGMA analysis showed consistent clustering of isolates; however, the fungus Leptospora rubella, which belongs to the order Dothideales, was grouped among species of the order Pleosporales. The presence of endophytic Fusarium species in G. max roots is unsurprising, since Fusarium spp. isolates have been previously described as endophyte in other reports. However, it remains to be determined whether the G. max Fusarium endophytes are latent pathogens or non-pathogenic forms that benefit the plant. This study provides a broader knowledge of the distribution of the fungal community in G. max leaves and roots, and identifies the genetic relationships among the isolated species. PMID:26111593

  18. Screening for Bacillus Isolates in the Broiler Gastrointestinal Tract

    PubMed Central

    Barbosa, Teresa M.; Serra, Cláudia R.; La Ragione, Roberto M.; Woodward, Martin J.; Henriques, Adriano O.

    2005-01-01

    Spores from a number of different Bacillus species are currently being used as human and animal probiotics, although their mechanisms of action remain poorly understood. Here we describe the isolation of 237 presumptive gut-associated Bacillus spp. isolates that were obtained by heat and ethanol treatment of fecal material from organically reared broilers followed by aerobic plating. Thirty-one representative isolates were characterized according to their morphological, physiological, and biochemical properties as well as partial 16S rRNA gene sequences and screening for the presence of plasmid DNA. The Bacillus species identified included B. subtilis, B. pumilus, B. licheniformis, B. clausii, B. megaterium, B. firmus, and species of the B. cereus group, whereas a number of our isolates could not be classified. Intrinsic properties of potential importance for survival in the gut that could be advantageous for spore-forming probiotics were further investigated for seven isolates belonging to five different species. All isolates sporulated efficiently in the laboratory, and the resulting spores were tolerant to simulated gastrointestinal tract conditions. They also exhibited antimicrobial activity against a broad spectrum of bacteria, including food spoilage and pathogenic organisms such as Bacillus spp., Clostridium perfringens, Staphylococcus aureus, and Listeria monocytogenes. Importantly, the isolates were susceptible to most of the antibiotics tested, arguing that they would not act as donors for resistance determinants if introduced in the form of probiotic preparations. Together, our results suggest that some of the sporeformers isolated in this study have the potential to persist in or transiently associate with the complex gut ecosystem. PMID:15691955

  19. Covalent immobilization of xylanase produced from Bacillus pumilus SV-85S on electrospun polymethyl methacrylate nanofiber membrane.

    PubMed

    Kumar, Pankaj; Gupta, Ashish; Dhakate, Sanjay R; Mathur, Rakesh B; Nagar, Sushil; Gupta, Vijay K

    2013-01-01

    Polymethyl methacrylate (PMMA) nanofiber membrane (NFM) was synthesized by an electrospinning technique. These membranes were utilized as a support for immobilization of xylanase enzyme to study its pH stability, thermal stability, and reusability. The morphology of aligned NFM was studied by optical microscopy and scanning electron microscopy. The PMMA NFM was functionalized with phenylenediamine and activated with glutaraldehyde to yield an aldehyde group on its surface for covalent immobilization of xylanase. The Fourier transform infrared analysis of the covalently immobilized xylanase confirmed that the enzyme was immobilized on PMMA NFM via amide linkages. The immobilization efficiency of covalently bound xylanase was found experimentally to be 90%. A forward shift in pH optima from 6.0-7.0 (soluble enzyme) to 7.0-9.0 (immobilized enzyme) was observed after xylanase immobilization. The pH and temperature stability of xylanase were enhanced upon its covalent immobilization. The immobilized enzyme was active on repeated use and retained ?80% of its initial activity after 11 reaction cycles. The improved thermal and operational stability of the covalently immobilized enzyme on PMMA NFM might be advantageous for industrial applications. PMID:23586605

  20. Microarray analysis of Endophyte-infected and Endophyte-free tall fescue

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Many grasses have mutualistic symbioses with fungi of the family Clavicipitaceae. Tall fescue [Lolium arundinaceum (Schreb.) = Schedonorus arundinaceus (Schreb.) Dumont.] can harbor the obligate endophyte, Neotyphodium coenophialum that is asexually propagated and transmitted via host seeds. Total...

  1. Bioactivity of root endophytic freshwater Hyphomycetes Anguillospora longissima (Sacc. & Syd.) Ingold.

    PubMed

    Sati, S C; Singh, Lokendra

    2014-01-01

    Anguillospora longissima, isolated from root as endophytic freshwater hyphomycetes, was evaluated for its bioactivity (antibacterial potential) against five bacterial strains, namely, Gram-positive (Bacillus subtilis MTCC 121) and Gram-negative (Agrobacterium tumefaciens MTCC 609, Escherichia coli MTCC 40, Erwinia chrysanthemum, and Xanthomonas pseudomonas). Antimicrobial activity was assessed by measuring the zone of inhibition with preliminary and secondary antimicrobial assays. The applied fungus was found significant for all tested bacterial strains as showen by their zone of inhibition. In preliminary antimicrobial assay, maximum zone of inhibition was recorded against Gram-negative human pathogenic bacterial strain Escherichia coli (23 mm) followed by Erwinia chrysanthemi (22 mm), Agrobacterium tumefaciens (21 mm), and Xanthomonas phaseoli (21 mm), while minimum zone of inhibition was observed against Bacillus subtilis (20 mm). In secondary antimicrobial assay, the maximum zone of inhibition was recorded against Erwinia chrysanthemi (11 mm) followed by Agrobacterium tumefaciens (10 mm), Xanthomonas phaseoli (10 mm), and Bacillus subtilis (9 mm) and minimum inhibition was found against Escherichia coli (8 mm). PMID:25383378

  2. Isolation of potential probiotic Bacillus spp. and assessment of their subcellular components to induce immune responses in Labeo rohita against Aeromonas hydrophila.

    PubMed

    Ramesh, Dharmaraj; Vinothkanna, Annadurai; Rai, Amit Kumar; Vignesh, Venkada Subramanian

    2015-08-01

    Bacillus species isolated from the gut of healthy Labeo rohita (Hamilton) were screened for antibacterial activity against selected fish pathogens. Among the isolates, KADR5 and KADR6 showed antibacterial activity, tolerated low pH and high bile concentrations and were susceptibility to various antibiotics. Based on morphological and biochemical tests and 16S rRNA gene analysis the probiotic strains KADR5 and KADR6 were identified as Bacillus licheniformis and Bacillus pumilus, respectively. The immune stimulatory effect of subcellular components of probiotic Bacillus licheniformis KADR5 and Bacillus pumilus KADR6 in L. rohita against Aeromonas hydrophila infection was studied. Fish were immunized intraperitoneally in case of subcellular components [cell wall proteins (CWPs), extracellular proteins (ECPs), whole cell proteins (WCPs)] and orally in case of live cells (10(8) CFU/g of feed). After 14th day of administration, fishes from each group were challenged intraperitoneally with 0.1 ml of A. hydrophila cell suspension in PBS (10(5) cells ml(-1)). Groups immunized with subcellular components and live cells had significantly lower mortalities of 20-40% and 23-33%, respectively in comparison to control (80% mortality). The non specific immune factors in the cellular components and viable cells of the probiotics increased the expression of lysozyme and respiratory burst. Use of WCPs and CWPs resulted in better protection against A. hydrophila in L. rohita. Our results clearly reflect the potential of cellular components of the probiotics Bacillus species for the protection of fish against A. hydrophila infection by enhancing the immune response. PMID:25917974

  3. Endophytic bacterial diversity in grapevine (Vitis vinifera L.) leaves described by 16S rRNA gene sequence analysis and length heterogeneity-PCR.

    PubMed

    Bulgari, Daniela; Casati, Paola; Brusetti, Lorenzo; Quaglino, Fabio; Brasca, Milena; Daffonchio, Daniele; Bianco, Piero Attilio

    2009-08-01

    Diversity of bacterial endophytes associated with grapevine leaf tissues was analyzed by cultivation and cultivation-independent methods. In order to identify bacterial endophytes directly from metagenome, a protocol for bacteria enrichment and DNA extraction was optimized. Sequence analysis of 16S rRNA gene libraries underscored five diverse Operational Taxonomic Units (OTUs), showing best sequence matches with gamma-Proteobacteria, family Enterobacteriaceae, with a dominance of the genus Pantoea. Bacteria isolation through cultivation revealed the presence of six OTUs, showing best sequence matches with Actinobacteria, genus Curtobacterium, and with Firmicutes genera Bacillus and Enterococcus. Length Heterogeneity-PCR (LH-PCR) electrophoretic peaks from single bacterial clones were used to setup a database representing the bacterial endophytes identified in association with grapevine tissues. Analysis of healthy and phytoplasma-infected grapevine plants showed that LH-PCR could be a useful complementary tool for examining the diversity of bacterial endophytes especially for diversity survey on a large number of samples. PMID:19763412

  4. The secret world of endophytes in perspective

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This work in Fungal Ecology is focused on the group of plant symbionts that have been termed collectively ‘microbial endophytes’. Broadly, microbial endophytes are commonly considered to be any of a diverse group of bacteria, cyanobacteria, or fungi that colonize internal tissues of plants. After ...

  5. Sebacinales Everywhere: Previously Overlooked Ubiquitous Fungal Endophytes

    PubMed Central

    Weiß, Michael; Sýkorová, Zuzana; Garnica, Sigisfredo; Riess, Kai; Martos, Florent; Krause, Cornelia; Oberwinkler, Franz; Bauer, Robert; Redecker, Dirk

    2011-01-01

    Inconspicuous basidiomycetes from the order Sebacinales are known to be involved in a puzzling variety of mutualistic plant-fungal symbioses (mycorrhizae), which presumably involve transport of mineral nutrients. Recently a few members of this fungal order not fitting this definition and commonly referred to as ‘endophytes’ have raised considerable interest by their ability to enhance plant growth and to increase resistance of their host plants against abiotic stress factors and fungal pathogens. Using DNA-based detection and electron microscopy, we show that Sebacinales are not only extremely versatile in their mycorrhizal associations, but are also almost universally present as symptomless endophytes. They occurred in field specimens of bryophytes, pteridophytes and all families of herbaceous angiosperms we investigated, including liverworts, wheat, maize, and the non-mycorrhizal model plant Arabidopsis thaliana. They were present in all habitats we studied on four continents. We even detected these fungi in herbarium specimens originating from pioneering field trips to North Africa in the 1830s/40s. No geographical or host patterns were detected. Our data suggest that the multitude of mycorrhizal interactions in Sebacinales may have arisen from an ancestral endophytic habit by specialization. Considering their proven beneficial influence on plant growth and their ubiquity, endophytic Sebacinales may be a previously unrecognized universal hidden force in plant ecosystems. PMID:21347229

  6. Acid protease production in fungal root endophytes.

    PubMed

    Mayerhofer, Michael S; Fraser, Erica; Kernaghan, Gavin

    2015-01-01

    Fungal endophytes are ubiquitous in healthy root tissue, but little is known about their ecosystem functions, including their ability to utilize organic nutrient sources such as proteins. Root-associated fungi may secrete proteases to access the carbon and mineral nutrients within proteins in the soil or in the cells of their plant host. We compared the protein utilization patterns of multiple isolates of the root endophytes Phialocephala fortinii s.l., Meliniomyces variabilis and Umbelopsis isabellina with those of two ectomycorrhizal (ECM) fungi, Hebeloma incarnatulum and Laccaria bicolor, and the wood-decay fungus Irpex lacteus at pH values of 2-9 on liquid BSA media. We also assessed protease activity using a fluorescently labeled casein assay and gelatin zymography and characterized proteases using specific protease inhibitors. I. lacteus and U. isabellina utilized protein efficiently, while the ECM fungi exhibited poor protein utilization. ECM fungi secreted metallo-proteases and had pH optima above 4, while other fungi produced aspartic proteases with lower pH optima. The ascomycetous root endophytes M. variabilis and P. fortinii exhibited intermediate levels of protein utilization and M. variabilis exhibited a very low pH optimum. Comparing proteolytic profiles between fungal root endophytes and fungi with well defined ecological roles provides insight into the ecology of these cryptic root associates. PMID:25344260

  7. Rice Plants Grown With and Without Endophytes

    USGS Multimedia Gallery

    These rice plants show the difference in growth of rice plants exposed to salt when grown with and without endophytes, which are mutually beneficial microscopic fungi that live in most plants. The plant on the left was colonized with a fungi that made it salt-tolerant, but it wasn't exposed to ...

  8. Fungal endophytes in green coffee seeds

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Green coffee seeds from Colombia, Guatemala, India, Kenya, Papua New Guinea, Puerto Rico, and Vietnam were sampled for the presence of fungal endophytes. Sections of surface sterilized seeds were plated on yeast malt agar, and fungal growth was isolated for subsequent DNA extraction and sequencing....

  9. Endophytic bacteria from Piper tuberculatum Jacq.: isolation, molecular characterization, and in vitro screening for the control of Fusarium solani f. sp piperis, the causal agent of root rot disease in black pepper (Piper nigrum L.).

    PubMed

    Nascimento, S B; Lima, A M; Borges, B N; de Souza, C R B

    2015-01-01

    Endophytic bacteria have been found to colonize internal tissues in many different plants, where they can have several beneficial effects, including defense against pathogens. In this study, we aimed to identify endophytic bacteria associated with roots of the tropical piperaceae Piper tuberculatum, which is known for its resistance to infection by Fusarium solani f. sp piperis, the causal agent of black pepper (Piper nigrum) root rot disease in the Amazon region. Based on 16S rRNA gene sequence analysis, we isolated endophytes belonging to 13 genera: Bacillus, Paenibacillus, Pseudomonas, Enterobacter, Rhizobium, Sinorhizobium, Agrobacterium, Ralstonia, Serratia, Cupriavidus, Mitsuaria, Pantoea, and Staphylococcus. The results showed that 56.52% of isolates were associated with the phylum Proteobacteria, which comprised ?, ?, and ? classes. Other bacteria were related to the phylum Firmicutes, including Bacillus, which was the most abundant genus among all isolates. Antagonistic assays revealed that Pt12 and Pt13 isolates, identified as Pseudomonas putida and Pseudomonas sp, respectively, were able to inhibit F. solani f. sp piperis growth in vitro. We describe, for the first time, the molecular identification of 23 endophytic bacteria from P. tuberculatum, among which two Pseudomonas species have the potential to control the pathogen responsible for root rot disease in black pepper in the Amazon region. PMID:26214435

  10. Characterization of Bacillus spp. strains for use as probiotic additives in pig feed.

    PubMed

    Larsen, Nadja; Thorsen, Line; Kpikpi, Elmer Nayra; Stuer-Lauridsen, Birgitte; Cantor, Mette Dines; Nielsen, Bea; Brockmann, Elke; Derkx, Patrick M F; Jespersen, Lene

    2014-02-01

    Bacillus spp. are commonly used as probiotic species in the feed industry, however, their benefits need to be confirmed. This study describes a high throughput screening combined with the detailed characterization of endospore-forming bacteria with the aim to identify new Bacillus spp. strains for use as probiotic additives in pig feed. A total of 245 bacterial isolates derived from African fermented food, feces and soil were identified by 16S rRNA gene sequencing and screened for antimicrobial activity and growth in the presence of antibiotics, bile salts and at pH 4.0. Thirty-three Bacillus spp. isolates with the best characteristics were identified by gyrB and rpoB gene sequencing as B. amyloliquefaciens subsp. plantarum, B. amyloliquefaciens subsp. amyloliquefaciens, B. subtilis subsp. subtilis, B. licheniformis, B. mojavensis, B. pumilus and B. megaterium. These isolates were further investigated for their activity against the pathogenic bacteria, antibiotic susceptibility, sporulation rates, biofilm formation and production of glycosyl hydrolytic enzymes. Additionally, ten selected isolates were assessed for heat resistance of spores and the effect on porcine epithelial cells IPEC-J2. Isolates of B. amyloliquefaciens, B. subtilis and B. mojavensis, showed the best overall characteristics and, therefore, potential for usage as probiotic additives in feed. A large number of taxonomically diverse strains made it possible to reveal species and subspecies-specific trends, contributing to our understanding of the probiotic potential of Bacillus species. PMID:24201893

  11. [Distribution and characteristics of Bacillus bacteria associated with hydrobionts and the waters of the Peter the Great Bay, Sea of Japan].

    PubMed

    Beleneva, I A

    2008-01-01

    Bacilli of the species Bacillus subtilis, B. pumilus, B. mycoides, B. marinus and B. licheniformis (a total of 53 strains) were isolated from 15 invertebrate species and the water of the Vostok Bay, Peter the Great Bay, Sea of Japan. Bacilli were most often isolated from bivalves (22.7%) and sea cucumbers (18.9%); they occurred less frequently in sea urchins and starfish (13.2 and 7.5%, respectively). Most of bacilli strains were isolated from invertebrates inhabiting silted sediments. No Bacillus spp. strains were isolated from invertebrates inhabiting stony and sandy environments. The species diversity of bacilli isolated from marine objects under study was low. Almost all bacterial isolates were resistant to lincomycin. Unlike B. pumilus, B. subtilis isolates were mostly resistant to benzylpenicillin and ampicillin. Antibiotic sensitivity of B. licheniformis strains was variable (two strains were resistant to benzylpenicillin and oxacillin, while one was sensitive). A significant fraction of isolated bacilli contained pigments. Pigmented strains were more often isolated from seawater samples, while colorless ones predominated within hydrobionts. B. subtilis colonies had the broadest range of colors. In the Bacillus strains obtained, DNase, RNase, phosphatase, elastolytic, chitinase, and agarolytic activity was detected. Bacilli strains with hydrolytic activity occurred in invertebrates more often than in seawater. PMID:18825984

  12. Plants and endophytes: equal partners in secondary metabolite production?

    PubMed

    Ludwig-Müller, Jutta

    2015-07-01

    Well known plant production systems should be re-evaluated due to findings that the interesting metabolite might actually be produced by microbes intimately associated with the plant, so-called endophytes. Endophytes can be bacteria or fungi and they are characterized usually by the feature that they do not cause any harm to the host. Indeed, in some cases, such as mycorrhizal fungi or other growth promoting endophytes, they can be beneficial for the plant. Here some examples are reviewed where the host plant and/or endophyte metabolism can be induced by the other partner. Also, partial or complete biosynthesis pathways for plant secondary metabolites can be attributed to such endophytes. In other cases the host plant is able to metabolize substances from fungal origin. The question of the natural role of such metabolic changes for the endophyte will be briefly touched. Finally, the consequences for the use of plant cultures for secondary metabolite production is discussed. PMID:25792513

  13. Characteristics and biodiversity of endophytic phosphorus- and potassium-solubilizing bacteria in Moso Bamboo (Phyllostachys edulis).

    PubMed

    Yuan, Zong-Sheng; Liu, Fang; Zhang, Guo-Fang

    2015-12-01

    Endophytic phosphorus- and potassium-solubilizing bacteria were screened from the root, rhizome, stem, and leaves of Moso Bamboo, and their diversity was analyzed using their 16S rDNA sequences. Twenty endophytic phosphorus and potassium-solubilizing bacteria were screened from 82 bamboo plants, among which the CT-B09-2, WYS-A01-1 and JL-B06 had higher activities in decomposing organophosphates. The three species showed a decomposition diameter/colony diameter (D/d) of 5.05, 4.19 and 2.95, respectively, and a solubilizing activity of 81.77 mg/L, 77.85 mg/L and 63.69 mg/L, respectively. JL-B06, WYS-A01-1 and CT-B09-2 had higher activities in decomposing inorganic phosphorus, with a decomposition diameter/colony diameter (D/d) of 2.34, 2.12 and 1.82, respectively, and a solubilizing activity of 30.58 mg/L, 38.89 mg/L and 48.35 mg/L, respectively. CT-B21, WYS-A03-1 and JL-B06 had higher activities in decomposing potassium, with a decomposition diameter/colony diameter (D/d) of 3.37, 4.84 and 4.33, respectively, and a solubilizing activity of 2.81 mg/L, 2.54 mg/L and 2.46 mg/L, respectively. The 16S rDNA sequence analysis showed that the 20 phosphorus- and potassium-solubilizing bacteria belong to 14 species from 10 genera, and mainly consist of Alcaligenes spp., Enterobacter spp. and Bacillus spp. Our results demonstrate the abundant diversity of endophytic phosphorus- and potassiumsolubilizing bacteria in Moso Bamboo. PMID:26616376

  14. Developmental Peculiarities and Seed-Borne Endophytes in Quinoa: Omnipresent, Robust Bacilli Contribute to Plant Fitness.

    PubMed

    Pitzschke, Andrea

    2016-01-01

    Among potential climate change-adapted crops for future agriculture, quinoa (Chenopodium quinoa), a facultative halophyte plant with exceptional nutritional properties, stands out as a prime candidate. This work examined how quinoa deals with extreme situations during seed rehydration. Quinoa distinguishes itself from other plants in multiple ways. It germinates within minutes, even under extremely hostile conditions. Broken seeds/split embryos are able to regenerate. Furthermore, quinoa seedlings are resurrection-competent. These peculiarities became in part explainable upon discovery of seed-borne microorganisms. 100% of quinoa seeds, from different sources, are inhabited by diverse members of the genus Bacillus. These endophytes are motile and reside in all seedling organs, indicating vertical transmission. Owing to their high catalase activities and superoxide contents the bacteria potentially manipulate the host's redox status. Superoxide-driven cell expansion enables quinoa to overcome a critical period in development, seedling establishment. Quinoa's immediate confrontation with "foreign" reactive oxygen species and bacterial elicitors likely induces a naturally primed state, enabling plants to withstand extreme situations. The endophytic bacteria, which are cultivable and highly robust themselves, have high potential for application in agriculture, food (amylase) and cosmetics (catalase) industry. This work also discusses the potential of transferring quinoa's microbiome to improve stress resistance in other plant species. PMID:26834724

  15. Developmental Peculiarities and Seed-Borne Endophytes in Quinoa: Omnipresent, Robust Bacilli Contribute to Plant Fitness

    PubMed Central

    Pitzschke, Andrea

    2016-01-01

    Among potential climate change-adapted crops for future agriculture, quinoa (Chenopodium quinoa), a facultative halophyte plant with exceptional nutritional properties, stands out as a prime candidate. This work examined how quinoa deals with extreme situations during seed rehydration. Quinoa distinguishes itself from other plants in multiple ways. It germinates within minutes, even under extremely hostile conditions. Broken seeds/split embryos are able to regenerate. Furthermore, quinoa seedlings are resurrection-competent. These peculiarities became in part explainable upon discovery of seed-borne microorganisms. 100% of quinoa seeds, from different sources, are inhabited by diverse members of the genus Bacillus. These endophytes are motile and reside in all seedling organs, indicating vertical transmission. Owing to their high catalase activities and superoxide contents the bacteria potentially manipulate the host’s redox status. Superoxide-driven cell expansion enables quinoa to overcome a critical period in development, seedling establishment. Quinoa’s immediate confrontation with “foreign” reactive oxygen species and bacterial elicitors likely induces a naturally primed state, enabling plants to withstand extreme situations. The endophytic bacteria, which are cultivable and highly robust themselves, have high potential for application in agriculture, food (amylase) and cosmetics (catalase) industry. This work also discusses the potential of transferring quinoa’s microbiome to improve stress resistance in other plant species. PMID:26834724

  16. Polyketide and benzopyran compounds of an endophytic fungus isolated from Cinnamomum mollissimum: biological activity and structure

    PubMed Central

    Santiago, Carolina; Sun, Lin; Munro, Murray Herbert Gibson; Santhanam, Jacinta

    2014-01-01

    Objective To study bioactivity and compounds produced by an endophytic Phoma sp. fungus isolated from the medicinal plant Cinnamomum mollissimum. Methods Compounds produced by the fungus were extracted from fungal broth culture with ethyl acetate. This was followed by bioactivity profiling of the crude extract fractions obtained via high performance liquid chromatography. The fractions were tested for cytotoxicity to P388 murine leukemic cells and antimicrobial activity against bacteria and pathogenic fungi. Compounds purified from active fractions which showed antibacterial, antifungal and cytotoxic activities were identified using capillary nuclear magnetic resonance analysis, mass spectrometry and admission to AntiMarin database. Results Three known compounds, namely 4-hydroxymellein, 4,8-dihydroxy-6-methoxy-3-methyl-3,4-dihydro-1H-isochromen-1-one and 1-(2,6-dihydroxyphenyl) ethanone, were isolated from the fungus. The polyketide compound 4-hydroxymellein showed high inhibitory activity against P388 murine leukemic cells (94.6%) and the bacteria Bacillus subtilis (97.3%). Meanwhile, 4,8-dihydroxy-6-methoxy-3-methyl-3,4-dihydro-1H-isochromen-1-one, a benzopyran compound, demonstrated moderate inhibitory activity against P388 murine leukemic cells (48.8%) and the fungus Aspergillus niger (56.1%). The second polyketide compound, 1 (2,6-dihydroxyphenyl) ethanone was inactive against the tested targets. Conclusions These findings demonstrate the potential of endophytes as producers of pharmacologically important compounds, including polyketides which are major secondary metabolites in fungi. PMID:25183332

  17. [Screening and identification of indoleacetic acid producing endophytic bacterium in Panax ginseng].

    PubMed

    Jiang, Yun; Tian, Lei; Chen, Chang-qing; Zhang, Guan-jun; Li, Tong; Chen, Jing-xiu; Wang, Xue

    2015-01-01

    Endophytic bacteria which was producing indoleacetic acid was screened from Panax ginseng by using the Salkowski method. The active strain was also tested for its ability of nitrogen fixation by using the Ashby agar plates, the PKV plates and quantitative analysis of Mo-Sb-Ascrobiology acid colorimetry was used to measure its ability of phosphate solubilization, for its ability of potassium solubilization the silicate medium and flame spectrophotometry was used, for its ability of producing siderophores the method detecting CAS was used, for its ability of producing ACC deaminase the Alpha ketone butyric acid method was applied. And the effect on promoting growth of seed by active strain was tested. The results showed that the indoleacetic acid producing strain of JJ5-2 was obtained from 118 endophytes, which the content of indoleacetic acid was 10.2 mg x L(-1). The JJ5-2 strain also had characteristics of phosphate and potassium solubilization, nitrogen fixation, producing siderophores traits, and the promoting germination of ginseng seeds. The JJ5-2 strain was identified as Bacillus thuringiensis by analyzing morphology, physiological and biochemical properties and 16S rRNA gene sequences. PMID:26080547

  18. Bacterial endophytes contribute to abiotic stress adaptation in pepper plants (Capsicum annuum L.).

    PubMed

    Sziderics, A H; Rasche, F; Trognitz, F; Sessitsch, A; Wilhelm, E

    2007-11-01

    Endophytes are nonpathogenic plant-associated bacteria that can play an important role in plant vitality and may confer resistance to abiotic or biotic stress. The effects of 5 endophytic bacterial strains isolated from pepper plants showing 1-aminocyclopropane-1-carboxylate deaminase activity were studied in sweet pepper under in vitro conditions. Four of the strains tested showed production of indole acetic acid. Plant growth, osmotic potential, free proline content, and gene expression were monitored in leaves and roots under control and mild osmotic stress conditions. All indole acetate producers promoted growth in Capsicum annuum L. 'Ziegenhorn Bello', from which they were isolated. Osmotic stress caused an increase in the content of free proline in the leaves of both inoculated and noninoculated plants. Inoculated control plants also revealed higher proline levels in comparison with noninoculated control plants. Differential gene expression patterns of CaACCO, CaLTPI, CaSAR82A, and putative P5CR and P5CS genes during moderate stress were observed, depending on the bacterium applied. Inoculation with 2 bacterial strains, EZB4 and EZB8 (Arthrobacter sp. and Bacillus sp., respectively), resulted in a significantly reduced upregulation or even downregulation of the stress-inducible genes CaACCO and CaLTPI, as compared with the gene expression in noninoculated plants. This indicates that both strains reduced abiotic stress in pepper under the conditions tested. PMID:18026213

  19. Identification of bacterial endophytes associated with traditional medicinal plant Tridax procumbens Linn.

    PubMed Central

    Preveena, Jagadesan; Bhore, Subhash J.

    2013-01-01

    Background: In traditional medicine, Tridax procumbens Linn. is used in the treatment of injuries and wounds. The bacterial endophytes (BEs) of medicinal plants could produce medicinally important metabolites found in their hosts; and hence, the involvement of BEs in conferring wound healing properties to T. Procumbens cannot be ruled out. But, we do not know which types of BEs are associated with T. Procumbens. Objective: The objective of this study was to investigate the fast growing and cultivable BEs associated with T. procumbens. Materials and Methods: Leaves and stems of healthy T. Procumbens plants were collected and cultivable BEs were isolated from surface-sterilized leaf and stem tissue samples using Luria-Bertani (LB) agar (medium) at standard conditions. A polymerase chain reaction was employed to amplify 16S rRNA coding gene fragments from the isolates. Cultivable endophytic bacterial isolates (EBIs) were identified using 16S rRNA gene nucleotide sequence similarity based method of bacterial identification. Results: Altogether, 50 culturable EBIs were isolated. 16S rRNA gene nucleotide sequences analysis using the Basic Local Alignment Search Tool (BLAST) revealed identities of the EBIs. Analysis reveals that cultivable Bacillus spp., Cronobacter sakazakii, Enterobacter spp., Lysinibacillus sphaericus, Pantoea spp., Pseudomonas spp. and Terribacillus saccharophilus are associated with T. Procumbens. Conclusion: Based on the results, we conclude that 24 different types of culturable BEs are associated with traditionally used medicinal plant, T. Procumbens, and require further study. PMID:24501447

  20. Structural and stereochemical studies of hydroxyanthraquinone derivatives from the endophytic fungus Coniothyrium sp.

    PubMed

    Sun, Peng; Huo, Juan; Kurtán, Tibor; Mándi, Attila; Antus, Sándor; Tang, Hua; Draeger, Siegfried; Schulz, Barbara; Hussain, Hidayat; Krohn, Karsten; Pan, Weihua; Yi, Yanghua; Zhang, Wen

    2013-02-01

    Four known hydroxyanthraquinones (1-4) together with four new derivatives having a tetralone moiety, namely coniothyrinones A-D (5-8), were isolated from the culture of Coniothyrium sp., an endophytic fungus isolated from Salsola oppostifolia from Gomera in the Canary Islands. The structures of the new compounds were elucidated by detailed spectroscopic analysis and comparison with reported data. The absolute configurations of coniothyrinones A (5), B (6), and D (8) were determined by TDDFT calculations of CD spectra, allowing the determination of the absolute configuration of coniothyrinone C (7) as well. Coniothyrinones A (5), B (6), and D (8) could be used as ECD reference compounds in the determination of absolute configuration for related tetralone derivatives. This is the first report of anthraquinones and derivatives from an isolate of the genus Coniothyrium sp. These compounds showed inhibitory effects against the fungus Microbotryum violaceum, the alga Chlorella fusca, and the bacteria Escherichia coli and Bacillus megaterium. PMID:23255384

  1. ROOT NODULE SYMBIOSIS. I. ENDOPHYTE OF MYRICA CERIFERA L.

    PubMed

    SILVER, W S

    1964-02-01

    Silver, Warren S. (University of Florida, Gainesville). Root nodule symbiosis. I. Endophyte of Myrica cerifera. J. Bacteriol. 87:416-421. 1964.-Electron microscopy of 0.1-mu thick sections of root nodules, fixed with permanganate and embedded with methacrylate, showed that infected plant cells were filled with a mycelial endophyte. The endophyte was filamentous, 1 mu in diameter, septate, and had an enlarged, club-shaped terminus. Although structurally the endophyte strongly resembles an actinomycete, it was not isolated in pure culture on a variety of appropriate media. PMID:14151065

  2. Poplar and its bacterial endophytes: coexistence and harmony

    SciTech Connect

    van der Lelie, D.; Taghavi, S.; Monchy, S.; Schwender, J.; Miller, L.; Ferrieri, R.; Rogers, A.; Zhu, W.; Weyens, N.; Vangronsveld, J.; Newman, L.

    2009-09-01

    Associations between plants and microorganisms are very complex and are the subject of an increasing number of studies. Here, we specifically address the relationship between poplar and its endophytic bacteria. The role and importance of endophytic bacteria in growth and development of their host plants is still underestimated. However, since many endophytes have a beneficial effect on their host, an improved understanding of the interaction between poplar and its endophytic bacteria has the potential to provide major breakthroughs that will improve the productivity of poplar. Endophytic bacteria can improve plant growth and development in a direct or indirect way. Direct plant growth promoting mechanisms may involve nitrogen fixation, production of plant growth regulators such as auxins, cytokinins and gibberellins, and suppression of stress ethylene synthesis by 1-aminocyclopropane-1-carboxylate (ACC) deaminase activity. Endophytic bacteria can indirectly benefit the plant by preventing the growth or activity of plant pathogens through competition for space and nutrients, antibiosis, production of hydrolytic enzymes, inhibition of pathogen-produced enzymes or toxins, and through systemic induction of plant defense mechanisms. Examples of applications for custom endophyte-host partnerships include improved productivity and establishment of poplar trees on marginal soils and the phytoremediation of contaminated soils and groundwater. A systems biology approach to understand the synergistic interactions between poplar and its beneficial endophytic bacteria represents an important field of research, which is facilitated by the recent sequencing of the genomes of poplar and several of its endophytic bacteria.

  3. Transcriptome response in different tissues of Lolium arundinaceum to the fungal endophyte Epichloe coenophiala

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Tall fescue (Lolium arundinaceum) plants symbiotic with the endophytic fungus, Epichloe coenophiala , (E+), have been shown to have better survivability and persistence than plants lacking the endophyte (E-). To understand more about the grass-endophyte interactions and how endophyte affects the ho...

  4. Endophytic fungus decreases plant virus infections in meadow ryegrass (Lolium pratense)

    PubMed Central

    Lehtonen, Päivi T; Helander, Marjo; Siddiqui, Shahid A; Lehto, Kirsi; Saikkonen, Kari

    2006-01-01

    We studied the effects of fungal endophyte infection of meadow ryegrass (Lolium pratense=Festuca pratensis) on the frequency of the barley yellow dwarf virus (BYDV). The virus is transferred by aphids, which may be deterred by endophyte-origin alkaloids within the plant. In our experiment, we released viruliferous aphid vectors on endophyte-infected and endophyte-free plants in a common garden. The number of aphids and the percentage of BYDV infections were lower in endophyte-infected plants compared to endophyte-free plants, indicating that endophyte infection may protect meadow ryegrass from BYDV infections. PMID:17148304

  5. Culture and Identification of Endophytic Fungi from Oxytropis glabra DC

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The Oxytropis glabra DC. is an important poisonous plant species in Inner Mongolia steppe. In this research the endophytic fungi from O. glabra DC. were studied both by microbiological and molecular biological techniques. The results showed as follows: The in vitro cultured endophytic fungi displ...

  6. Fungal endophyte increases the allelopathic effects of an invasive forb.

    PubMed

    Aschehoug, Erik T; Callaway, Ragan M; Newcombe, George; Tharayil, Nishanth; Chen, Shuyan

    2014-05-01

    Endophytic plant symbionts can have powerful effects on the way their hosts interact with pathogens, competitors, and consumers. The presence of endophytes in plants can alter food webs, community composition and ecosystem processes, suggesting that endophyte-plant symbioses may represent unique forms of extended phenotypes. We tested the impact of the fungal endophyte Alternaria alternata (phylotype CID 120) on the allelopathic effect of the invasive forb Centaurea stoebe when in competition with the North American native bunchgrass Koeleria macrantha in a greenhouse competition experiment. The allelopathic effect of C. stoebe on K. macrantha when infected with the fungal endophyte was more than twice that of endophyte-free C. stoebe. However, this allelopathic effect was a small part of the very large competitive effect of C. stoebe on K. macrantha in all treatments, likely because of the priority effects in our experimental design. To our knowledge, these results are the first experimental evidence for a symbiotic relationship between plants and fungal endophytes affecting allelopathic interactions between competing plants, and thus provide insight into the mechanisms by which fungal endophytes may increase the competitive ability of their hosts. PMID:24488227

  7. Endophyte status of tall fescue (festuca arundinacea) affects seed predation

    Technology Transfer Automated Retrieval System (TEKTRAN)

    In a preliminary study seed of a tall fescue (Festuca arundinacea Schreb.) variety ‘Jesup’ without endophyte were consumed at a slightly higher rate by common cricket (Acheta domesticus L.) in a standard feeding trial than the same fescue variety with the endophyte. Although, the preference for the...

  8. A novel method to scale up fungal endophyte isolations

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Estimations of species diversity are influenced by sampling intensity which in turn is influenced by methodology. For fungal endophyte diversity studies, the methodology includes surface-sterilization prior to isolation of endophytes. Surface-sterilization is an essential component of fungal endophy...

  9. Endophytic bacterial community of a Mediterranean marine angiosperm (Posidonia oceanica)

    PubMed Central

    Garcias-Bonet, Neus; Arrieta, Jesus M.; de Santana, Charles N.; Duarte, Carlos M.; Marbà, Núria

    2012-01-01

    Bacterial endophytes are crucial for the survival of many terrestrial plants, but little is known about the presence and importance of bacterial endophytes of marine plants. We conducted a survey of the endophytic bacterial community of the long-living Mediterranean marine angiosperm Posidonia oceanica in surface-sterilized tissues (roots, rhizomes, and leaves) by Denaturing Gradient Gel Electrophoresis (DGGE). A total of 26 Posidonia oceanica meadows around the Balearic Islands were sampled, and the band patterns obtained for each meadow were compared for the three sampled tissues. Endophytic bacterial sequences were detected in most of the samples analyzed. A total of 34 OTUs (Operational Taxonomic Units) were detected. The main OTUs of endophytic bacteria present in P. oceanica tissues belonged primarily to Proteobacteria (α, γ, and δ subclasses) and Bacteroidetes. The OTUs found in roots significantly differed from those of rhizomes and leaves. Moreover, some OTUs were found to be associated to each type of tissue. Bipartite network analysis revealed differences in the bacterial endophyte communities present on different islands. The results of this study provide a pioneering step toward the characterization of the endophytic bacterial community associated with tissues of a marine angiosperm and reveal the presence of bacterial endophytes that differed among locations and tissue types. PMID:23049528

  10. Geographic locality greatly influences fungal endophyte communities in Cephalotaxus harringtonia.

    PubMed

    Langenfeld, Aude; Prado, Soizic; Nay, Bastien; Cruaud, Corine; Lacoste, Sandrine; Bury, Edith; Hachette, François; Hosoya, Tsuyoshi; Dupont, Joëlle

    2013-02-01

    Although endophytes of conifers have been extensively studied, few data are available on Cephalotaxaceae. We examined foliar and stem endophytes of Cephalotaxus harringtonia, within its natural range in Japan and outside its natural range in France to study the effect of geography on endophyte community composition. In Japan, rapidly growing endophytes were dominant and may have masked the real diversity, in comparison to France where most endophytes were growing slowly. Analyses of ITS rDNA revealed 104 different Blast Groups among 554 isolates. Almost no overlap between endophyte assemblages of C. harringtonia from the two countries was observed. It seems that Japanese C. harringtonia trees, which should be well adapted to their native site, would host a specific, endemic endophyte community, while trees that have been introduced recently to a foreign site, in France, should have captured existing cosmopolitan and more generalist taxa. In Japan the majority of xylariaceous taxa, which dominated the communities, were unknown and, although closely related to Asian taxa, may be new to science. Dothideomycetes were more prevalent in France. Locally, urban environment, particularly in Japan, may have introduced some perturbations in the native endophyte community of C. harringtonia, with an abundance of generalist fungi such as Nigrospora and Colletotrichum. PMID:23452950

  11. 7 CFR 201.58d - Fungal endophyte test.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... aniline blue in 100 ml distilled water. (2) Prepare the endophyte staining solution of one part of 1... dilute with water if staining is too dark. (b) Procedure for determining levels of fungal endophyte in... thoroughly in running tap water. (4) De-glume seeds and place on a microscope slide in a drop of...

  12. 7 CFR 201.58d - Fungal endophyte test.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... aniline blue in 100 ml distilled water. (2) Prepare the endophyte staining solution of one part of 1... dilute with water if staining is too dark. (b) Procedure for determining levels of fungal endophyte in... thoroughly in running tap water. (4) De-glume seeds and place on a microscope slide in a drop of...

  13. 7 CFR 201.58d - Fungal endophyte test.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... aniline blue in 100 ml distilled water. (2) Prepare the endophyte staining solution of one part of 1... dilute with water if staining is too dark. (b) Procedure for determining levels of fungal endophyte in... thoroughly in running tap water. (4) De-glume seeds and place on a microscope slide in a drop of...

  14. 7 CFR 201.58d - Fungal endophyte test.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... aniline blue in 100 ml distilled water. (2) Prepare the endophyte staining solution of one part of 1... dilute with water if staining is too dark. (b) Procedure for determining levels of fungal endophyte in... thoroughly in running tap water. (4) De-glume seeds and place on a microscope slide in a drop of...

  15. 7 CFR 201.58d - Fungal endophyte test.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... aniline blue in 100 ml distilled water. (2) Prepare the endophyte staining solution of one part of 1... dilute with water if staining is too dark. (b) Procedure for determining levels of fungal endophyte in... thoroughly in running tap water. (4) De-glume seeds and place on a microscope slide in a drop of...

  16. Endophytic actinobacteria of medicinal plants: diversity and bioactivity.

    PubMed

    Golinska, Patrycja; Wypij, Magdalena; Agarkar, Gauravi; Rathod, Dnyaneshwar; Dahm, Hanna; Rai, Mahendra

    2015-08-01

    Endophytes are the microorganisms that exist inside the plant tissues without having any negative impact on the host plant. Medicinal plants constitute the huge diversity of endophytic actinobacteria of economical importance. These microbes have huge potential to synthesis of numerous novel compounds that can be exploited in pharmaceutical, agricultural and other industries. It is of prime importance to focus the present research on practical utilization of this microbial group in order to find out the solutions to the problems related to health, environment and agriculture. An extensive characterization of diverse population of endophytic actinobacteria associated with medicinal plants can provide a greater insight into the plant-endophyte interactions and evolution of mutualism. In the present review, we have discussed the diversity of endophytic actinobacteria of from medicinal plants their multiple bioactivities. PMID:26093915

  17. Antimicrobial activity of endophytic fungi isolated from Swietenia macrophylla leaves.

    PubMed

    Ibrahim, Darah; Lee, Chong Chai; Sheh-Hong, Lim

    2014-02-01

    The endophytic fungi isolated from leaves of Swietenia macrophylla of different ages were examined for antimicrobial activity. The agar plug diffusion assay was used for primary screening, followed by the disc diffusion method. A total of 461 filamentous endophytic fungi were isolated and cultured to examine their antimicrobial properties. In the primary screen, 315 isolates (68.3%) exhibited activity against at least one of the test pathogenic microorganisms. The percentage of isolates exhibiting antimicrobial activity increased with leaf age. Endophytic fungal assemblages, as well as those isolates exhibiting antimicrobial properties appeared to increase with leaf age. The main antimicrobial compounds were produced extracellularly by the endophytic fungi. The results suggest that healthy leaves at older stages of growth can be a potential source for the isolation of endophytic fungi with antimicrobial properties. PMID:24689302

  18. Evolutionary origins and ecological consequences of endophyte symbiosis with grasses.

    PubMed

    Clay, Keith; Schardl, Christopher

    2002-10-01

    Over the past 20 yr much has been learned about a unique symbiotic interaction between fungal endophytes and grasses. The fungi (Clavicipitaceae, Ascomycota) grow intercellularly and systemically in aboveground plant parts. Vertically transmitted asexual endophytes forming asymptomatic infections of cool-season grasses have been repeatedly derived from sexual species that abort host inflorescences. The phylogenetic distribution of seed-transmitted endophytes is strongly suggestive of cocladogenesis with their hosts. Molecular evidence indicates that many seed-transmitted endophytes are interspecific hybrids. Superinfection may result in hyphal fusion and parasexual recombination. Most endophytes produce one or more alkaloid classes that likely play some role in defending the host plant against pests. Hybridization may have led to the proliferation of alkaloid-production genes among asexual endophytes, favoring hybrids. The ergot alkaloid ergovaline, lolitrems, and lolines are produced by only a single sexual species, Epichloë festucae, but they are common in seed-transmitted endophytes, suggesting that E. festucae contributed genes for their synthesis. Asexual hybrids may also be favored by the counteracting of the accumulation of deleterious mutations (Muller's rachet). Endophyte infection can provide other benefits, such as enhanced drought tolerance, photosynthetic rate, and growth. Estimates of infection frequency have revealed variable levels of infection with especially high prevalence in the subfamily Pooideae. Longitudinal studies suggest that the prevalence of seed-transmitted endophytes can increase rapidly over time. In field experiments, infected tall fescue suppressed other grasses and forbs relative to uninfected fescue and supported lower consumer populations. Unlike other widespread plant/microbial symbioses based on the acquisition of mineral resources, grass/endophyte associations are based primarily on protection of the host from biotic and abiotic stresses. PMID:18707456

  19. Isolation and characterisation of Bacillus spp. antagonistic to Vibrio parahaemolyticus for use as probiotics in aquaculture.

    PubMed

    Liu, Xue-Fei; Li, Ya; Li, Jian-Rong; Cai, Lu-Yun; Li, Xiu-Xia; Chen, Jin-Ru; Lyu, Shu-Xia

    2015-05-01

    Acute gastroenteritis caused by pathogenic Vibrio parahaemolyticus is one of the major factors affecting the development of aquaculture and the safety of seafood. Using the antagonism of probiotics against pathogens is an alternative strategy to antibiotics and a common trend to control food-borne pathogenic bacteria. In this study, a total of 249 isolates were isolated from four types of seafood (Litopenaeus vannamei, Oratosquilla oratoria, Mactra veneriformis and Portunus trituberculatus) and coastal sediment from Liaodong Bay in the Bohai Sea, China with five different separation agars. The most isolates came from the sample of coastal sediment and on agar of 2216E, which accounted for 36.14 and 54.62 % respectively. Twenty-four among 249 isolates displayed direct antimicrobial activity to V. parahaemolyticus with spot inoculation. Sixteen active isolates were selected for extracellular antimicrobial activity using the Oxford cup method. Only strains of B16 and J7 showed extracellular antimicrobial activity and were identified as Bacillus pumilus and Bacillus mojavensis respectively based on the physiological identification and 16S rRNA sequence analysis. Both of the strains B16 and J7 exhibited extracellular hydrolytic enzyme activity and antagonism against more than one indicator bacteria in vitro, which indicates that the two strains have broad potential application as suitable probiotic candidates in aquaculture while B. mojavensis was first reported to inhibit pathogenic Vibrio spp. in vitro. There is no particular trait as to antagonism of B. pumilus B16 or B. mojavensis J7 to Gram-positive or Gram-negative indicator bacteria. PMID:25737203

  20. Efficacy of using harmless Bacillus endospores to estimate the inactivation of Cryptosporidium parvum oocysts in water.

    PubMed

    Garvey, Mary; Clifford, Eoghan; O'Reilly, Edmond; Rowan, Neil J

    2013-06-01

    The need to use complex in vitro cell culture, expensive equipment, and highly-trained technicians that are available only to specialist laboratories has significantly limited studies assessing the potential of pulsed UV light (PUV) to inactivate the waterborne parasite Cryptosporidium parvum in drinking water. This constitutes the first study to report on the use of different non-pathogenic Bacillus endospores as potential surrogate organisms to indicate the PUV inactivation performance of a C. parvum oocyst suspended in water. Findings showed that PUV effectively inactivated approximately 5 log10 CFU/ml Bacillus megaterium and Bacillus pumilus endospores suspended in water at a UV dose of 9.72 ?J/cm(2) that also inactivated statistically similar levels of C. parvum oocysts (P < 0.05), as determined by combined in vitro HCT-8 cell culture and quantitative PCR. Specifically, this study demonstrated that B. megaterium exhibited greater or similar PUV-inactivation kinetic data compared to that of similarly treated C. parvum over the UV dose range 6.4 to 12.9 ?J/cm(2). Therefore, the former may be used as an indicator organism for safely investigating the PUV-inactivation performance of this chlorine-resistant, waterborne parasite at the waste-water treatment plant level. Findings presented will impact positively on future water quality studies and on public health. PMID:23145570

  1. Automated ribotyping and antibiotic resistance determining of Bacillus spp from conjunctiva of diabetic patients

    PubMed Central

    Argun K?vanç, Sertaç; K?vanç, Merih; Güllülü, Gülay

    2014-01-01

    Objective(s): We aimed to characterize the phenotype and genotype of Bacillus spp isolated from diabetic patients’ eyes, by studying the drug sensitivity patterns with a disc-diffusion method. Materials and Methods: Fifty eyes of 25 patients with type II diabetes mellitus, with at least 10 years of diabetes history, were included in the study. We analyzed the eyes for the presence of Bacillus spp.; presumptive isolates were identified by morphological, and biochemical tests, and confirmed by the VITEK system. Automated EcoRI ribotyping was performed with a RiboPrinter® Microbial Characterization System. We determined the antibiotic resistance of the isolates by the Kirby–Bauer disc diffusion test. Results: Seven out of 25 patients were on insulin treatment; 7 on oral anti-diabetic medication; and 11 on combination therapy of insulin and oral medications. Among the 28 Bacillus spp isolates, 14 were B. cereus, 11 were B. pumilus, 2 were B. mojavensis and 1 was B. subtilis. Almost all the strains were either resistant or multiresistant, particularly towards cefuroxime, methicillin, and ceftazidime. Conclusion: Diabetic patients seem to be more prone to B. cereus infections than healthy individuals. It would be greatly beneficial to understand and recognize the prevalence of microorganisms and their resistance patterns for better outcome in ocular surgeries. PMID:24711899

  2. Probiotic applications of two dominant gut Bacillus strains with antagonistic activity improved the growth performance and immune responses of grouper Epinephelus coioides.

    PubMed

    Sun, Yun-Zhang; Yang, Hong-Ling; Ma, Ru-Long; Lin, Wen-Yan

    2010-11-01

    The effect of dietary administration of Bacillus pumilus and Bacillus clausii, the dominant bacteria with antagonistic activity in the gut of fast growing fish, on the growth performance and immune responses of grouper Epinephelus coioides were assessed. The fish were fed for 60 days with three different diets: control (without probiotics), diet T1 supplemented with 1.0x10(8) cells g(-1) B. pumilus, diet T2 with 1.0x10(8) cells g(-1) B. clausii. No significant improvements of weight gain or specific growth rate were observed in the probiotic fed groups, but a significant improvement of feed conversion ratio was observed after 60 days of feeding. Phagocytic activity and phagocytic index of fish fed probiotic diets were significantly higher than those of fish fed the control diet for 60 days. Superoxide dismutase (SOD) concentrations showed no significant difference between the treatments and the control during the whole experiment period, but which increased by 11.4% and 18.5% after 60 days of fed with diets T1 and T2, respectively. The serum lysozyme activities of fish fed diets T1 and T2 were significantly higher than that of fish fed control diet, and had respectively increased by 34.7% and 17.4% compared to the control after 60 days of feeding. Serum complement C3 levels of the treatments were significantly higher than that of control after 30 days of feeding, but no significant difference in serum complement C3 and C4 levels were observed between the treatments and the control after 60 days of feeding. The serum IgM levels of fish fed diet T1 and diet T2 were higher than that of fish fed control diet, and significant increase was observed in fish fed diet T2 for 30 days. The results demonstrated potential for B. pumilus and B. clausii to improve growth performance and immune responses of E. coioides. PMID:20637875

  3. Biocontrol potential of soybean bacterial endophytes against charcoal rot fungus, Rhizoctonia bataticola.

    PubMed

    Senthilkumar, M; Swarnalakshmi, K; Govindasamy, V; Lee, Young Keun; Annapurna, K

    2009-04-01

    A total of 137 bacterial isolates from surface sterilized root, stem, and nodule tissues of soybean were screened for their antifungal activity against major phytopathogens like Rhizoctonia bataticola, Macrophomina phaseolina, Fusarium udam, and Sclerotium rolfsii. Nine bacterial endophytes suppressed the pathogens under in vitro plate assay. These were characterized biochemically and identified at the genus level based on their partial sequence analysis of 16S rDNA. Eight of the isolates belonged to Bacillus and one to Paenibacillus. The phylogenetic relationship among the selected isolates was studied and phylogenetic trees were generated. The selected isolates were screened for biocontrol traits like production of hydrogen cyanide (HCN), siderophore, hydrolytic enzymes, antibiotics, and plant growth promoting traits like indole 3-acetic acid production, phosphate solubilization, and nitrogen fixation. A modified assessment scheme was used to select the most efficient biocontrol isolates Paenibacillus sp. HKA-15 (HKA-15) and Bacillus sp. HKA-121 (HKA-121) as potential candidates for charcoal rot biocontrol as well as soybean plant growth promotion. PMID:19067044

  4. Microarray Comparison of Endophyte-Infected and Endophyte-Free Tall Fescue

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Many grasses have mutualistic symbioses with fungi of the family Clavicipitaceae: Tall fescue (Festuca arudinacea = Lolium arundinaceum) can harbor the obligate endophyte, Neotyphodium coenophialum that are asexually propagated and transmitted via host seeds. In an effort to begin to dissect the hos...

  5. Performance by Spring-Calving Cows Grazing Tall Fescue Pastures with Either the Wild-Type Toxic Endophyte or a Non-Toxic Novel Endophyte

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Cows grazing 'Kentucky-31' tall fescue [Lolium arundinaceum (Schreb.) Darbysh.] infected with its wild-type endophyte (Neotyphodium coenophialum; E+) generally display suboptimal performance. Recently, endophyte strains that do not produce compounds toxic to cattle have been incorporated into tall ...

  6. Gene expression and metabolite analysis of endophyte infected and uninfected tall fescue clone pairs under water deficit conditions

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Tall fescue (Lolium arundinaceum) plants symbiotic with the endophytic fungus, Neotyphodium coenophialum (E+), have better survivability and persistence under stressful conditions, especially under drought stress, than plants lacking the endophyte (E-). To understand more about the grass-endophyte i...

  7. Assessment of the bacteriocinogenic potential of marine bacteria reveals lichenicidin production by seaweed-derived Bacillus spp.

    PubMed

    Prieto, Maria Luz; O'Sullivan, Laurie; Tan, Shiau Pin; McLoughlin, Peter; Hughes, Helen; O'Connor, Paula M; Cotter, Paul D; Lawlor, Peadar G; Gardiner, Gillian E

    2012-10-01

    The objectives of this study were (1) to assess the bacteriocinogenic potential of bacteria derived mainly from seaweed, but also sand and seawater, (2) to identify at least some of the bacteriocins produced, if any and (3) to determine if they are unique to the marine environment and/or novel. Fifteen Bacillus licheniformis or pumilus isolates with antimicrobial activity against at least one of the indicator bacteria used were recovered. Some, at least, of the antimicrobials produced were bacteriocins, as they were proteinaceous and the producers displayed immunity. Screening with PCR primers for known Bacillus bacteriocins revealed that three seaweed-derived Bacillus licheniformis harbored the bli04127 gene which encodes one of the peptides of the two-peptide lantibiotic lichenicidin. Production of both lichenicidin peptides was then confirmed by mass spectrometry. This is the first definitive proof of bacteriocin production by seaweed-derived bacteria. The authors acknowledge that the bacteriocin produced has previously been discovered and is not unique to the marine environment. However, the other marine isolates likely produce novel bacteriocins, as none harboured genes for known Bacillus bacteriocins. PMID:23170084

  8. Assessment of the Bacteriocinogenic Potential of Marine Bacteria Reveals Lichenicidin Production by Seaweed-Derived Bacillus spp.

    PubMed Central

    Luz Prieto, Maria; O’Sullivan, Laurie; Tan, Shiau Pin; McLoughlin, Peter; Hughes, Helen; O’Connor, Paula M.; Cotter, Paul D.; Lawlor, Peadar G.; Gardiner, Gillian E.

    2012-01-01

    The objectives of this study were (1) to assess the bacteriocinogenic potential of bacteria derived mainly from seaweed, but also sand and seawater, (2) to identify at least some of the bacteriocins produced, if any and (3) to determine if they are unique to the marine environment and/or novel. Fifteen Bacillus licheniformis or pumilus isolates with antimicrobial activity against at least one of the indicator bacteria used were recovered. Some, at least, of the antimicrobials produced were bacteriocins, as they were proteinaceous and the producers displayed immunity. Screening with PCR primers for known Bacillus bacteriocins revealed that three seaweed-derived Bacillus licheniformis harbored the bli04127 gene which encodes one of the peptides of the two-peptide lantibiotic lichenicidin. Production of both lichenicidin peptides was then confirmed by mass spectrometry. This is the first definitive proof of bacteriocin production by seaweed-derived bacteria. The authors acknowledge that the bacteriocin produced has previously been discovered and is not unique to the marine environment. However, the other marine isolates likely produce novel bacteriocins, as none harboured genes for known Bacillus bacteriocins. PMID:23170084

  9. Toxin-producing ability among Bacillus spp. outside the Bacillus cereus group.

    PubMed

    From, Cecilie; Pukall, Rudiger; Schumann, Peter; Hormazábal, Víctor; Granum, Per Einar

    2005-03-01

    A total of 333 Bacillus spp. isolated from foods, water, and food plants were examined for the production of possible enterotoxins and emetic toxins using a cytotoxicity assay on Vero cells, the boar spermatozoa motility assay, and a liquid chromatography-mass spectrometry method. Eight strains produced detectable toxins; six strains were cytotoxic, three strains produced putative emetic toxins (different in size from cereulide), and one strain produced both cytotoxin(s) and putative emetic toxin(s). The toxin-producing strains could be assigned to four different species, B. subtilis, B. mojavensis, B. pumilus, or B. fusiformis, by using a polyphasic approach including biochemical, chemotaxonomic, and DNA-based analyses. Four of the strains produced cytotoxins that were concentrated by ammonium sulfate followed by dialysis, and two strains produced cytotoxins that were not concentrated by such a treatment. Two cultures maintained full cytotoxic activity, two cultures reduced their activity, and two cultures lost their activity after boiling. The two most cytotoxic strains (both B. mojavensis) were tested for toxin production at different temperatures. One of these strains produced cytotoxin at growth temperatures ranging from 25 to 42 degrees C, and no reduction in activity was observed even after 24 h of growth at 42 degrees C. The strains that produced putative emetic toxins were tested for the influence of time and temperature on the toxin production. It was shown that they produced putative emetic toxin faster or just as fast at 30 as at 22 degrees C. None of the cytotoxic strains produced B. cereus-like enterotoxins as tested by PCR or by immunological methods. PMID:15746316

  10. Connecting plant-microbial interactions above and belowground: a fungal endophyte affects decomposition.

    PubMed

    Lemons, Alisha; Clay, Keith; Rudgers, Jennifer A

    2005-10-01

    Mutualisms can strongly affect the structure of communities, but their influence on ecosystem processes is not well resolved. Here we show that a plant-microbial mutualism affects the rate of leaf litter decomposition using the widespread interaction between tall fescue grass (Lolium arundinaceum) and the fungal endophyte Neotyphodium coenophialum. In grasses, fungal endophytes live symbiotically in the aboveground tissues, where the fungi gain protection and nutrients from their host and often protect host plants from biotic and abiotic stress. In a field experiment, decomposition rate depended on a complex interaction between the litter source (collected from endophyte-infected or endophyte-free plots), the decomposition microenvironment (endophyte-infected or endophyte-free plots), and the presence of mesoinvertebrates (manipulated by the mesh size of litter bags). Over all treatments, decomposition was slower for endophyte-infected fescue litter than for endophyte-free litter. When mesoinvertebrates were excluded using fine mesh and litter was placed in a microenvironment with the endophyte, the difference between endophyte-infected and endophyte-free litter was strongest. In the presence of mesoinvertebrates, endophyte-infected litter decomposed faster in microenvironments with the endophyte than in microenvironments lacking the endophyte, suggesting that plots differ in the detritivore assemblage. Indeed, the presence of the endophyte in plots shifted the composition of Collembola, with more Hypogastruridae in the presence of the endophyte and more Isotomidae in endophyte-free plots. In a separate outdoor pot experiment, we did not find strong effects of the litter source or the soil microbial/microinvertebrate community on decomposition, which may reflect differences between pot and field conditions or other differences in methodology. Our work is among the first to demonstrate an effect of plant-endophyte mutualisms on ecosystem processes under field conditions. PMID:16001218

  11. Phylogeny of marine Bacillus isolates from the Gulf of Mexico

    NASA Technical Reports Server (NTRS)

    Siefert, J. L.; Larios-Sanz, M.; Nakamura, L. K.; Slepecky, R. A.; Paul, J. H.; Moore, E. R.; Fox, G. E.; Jurtshuk, P. Jr

    2000-01-01

    The phylogeny of 11 pigmented, aerobic, spore-forming isolates from marine sources was studied. Forty-two biochemical characteristics were examined, and a 16S rDNA sequence was obtained for each isolate. In a phylogenetic tree based on 16S sequencing, four isolates (NRRL B-14850, NRRL B-14904, NRRL B-14907, and NRRL B-14908) clustered with B. subtilis and related organisms; NRRL B-14907 was closely related to B. amyloliquefaciens. NRRL B-14907 and NRRL B-14908 were phenotypically similar to B. amyloliquefaciens and B. pumilus, respectively. Three strains (NRRL B-14906, NRRL B-14910, and NRRL B-14911) clustered in a clade that included B. firmus, B. lentus, and B. megaterium. NRRL B-14910 was closely related phenotypically and phylogenetically to B. megaterium. NRRL B-14905 clustered with the mesophilic round spore-producing species, B. fusiformis and B. sphaericus; the isolate was more closely related to B. fusiformis. NRRL B-14905 displayed characteristics typical of the B. sphaericus-like organisms. NRRL B-14909 and NRRL B-14912 clustered with the Paenibacillus species and displayed characteristics typical of the genus. Only NRRL B-14851, an unusually thin rod that forms very small spores, may represent a new Bacillus species.

  12. Pogostol biosynthesis by the endophytic fungus Geniculosporium.

    PubMed

    Barra, Lena; Schulz, Barbara; Dickschat, Jeroen S

    2014-11-01

    Six (13)C-labelled isotopomers of mevalonolactone were synthesised and used in feeding experiments with the endophytic fungus Geniculosporium. The high incorporation rates of (13)C-label into a sesquiterpene that was found in headspace extracts of the fungus enabled unambiguous identification of this volatile as pogostol without the need for compound purification, simply by collecting the volatile fraction with a closed-loop stripping apparatus followed by direct (13)C NMR analysis (CLSA-NMR). The feeding experiments also gave insights into the biosynthesis of pogostol, including stereochemical aspects of the terpene cyclisation reaction. The possible biological function of pogostol is discussed. PMID:25186118

  13. Endophyte-mediated resistance to black cutworm as a function of plant cultivar and endophyte strain in tall fescue.

    PubMed

    Baldauf, Michael W; Mace, Wade J; Richmond, Douglas S

    2011-06-01

    To improve Neotyphodium endophyte-mediated resistance to black cutworm Agrotis ipsilon (Hufnagel) (BCW), a series of experiments was conducted by using several different cultivars of tall fescue, Schedonorus arundinaceus (Schreb.) Dumort. in combination with several different haplotypes of the endophyte Neotyphodium coenophialum (Morgan-Jones & Gams) (plant cultivar × endophyte haplotype = plant line), each producing unique alkaloid profiles. BCW settling response, survival at 5 and 10 d, and larval biomass varied significantly among plant lines. In general, greater variation BCW performance was observed within a single plant cultivar infected with different endophyte haplotypes than among different plant cultivars infected with the same endophyte haplotype, but comparisons among the former were far more numerous. Although five endophyte-mediated alkaloids representing three alkaloid classes were quantified in the plants, the pyrrolizidine alkaloid N-acetyl norloline was consistently the single best predictor of BCW performance. BCW settling response, 5-d survival, and 10-d survival decreased as levels of the alkaloid N-acetyl norloline increased. The same three response variables also decreased with increasing levels of peramine, but increased with increasing levels of ergovaline. Minor variation in endophyte infection levels occurring among infected plant lines had no significant influence on BCW performance. Results indicate a potentially important role for N-acetyl norloline and peramine in providing resistance to black cutworm whereas ergovaline appears to be much less important. Therefore, endophyte haplotypes expressing high levels of N-acetyl norloline and peramine may be of particular importance for developing 'friendly' endophyte-enhanced turf and pasture grasses that resist challenging lepidopteran pests, although remaining safe for wildlife and grazing mammals. PMID:22251642

  14. Diversity of bacterial endophytes in 3 and 15 year-old grapevines of Vitis vinifera cv. Corvina and their potential for plant growth promotion and phytopathogen control.

    PubMed

    Andreolli, Marco; Lampis, Silvia; Zapparoli, Giacomo; Angelini, Elisa; Vallini, Giovanni

    2016-02-01

    This study represents the first investigation on ecology of endophytic bacteria isolated from 3 and 15 year-old vine stems of Vitis vinifera cv. Corvina. The analysis was performed by means of culture-dependent techniques. The obtained results showed that new grapevine endophytic genera are being discovered. Moreover, Bacilli and Actinobacteria are frequently isolated from 3 year-old plants, whereas Alpha- and Gamma- Proteobacteria classes are more prevalent in the 15 year-old plants. Shannon-Wiener (H) index and analysis of rarefaction curves revealed greater genus richness in young grapevine plants. Furthermore, results evidenced an increase of genotypic group number within specific genera (e.g., Rhizobium and Pantoea). Among isolated strains from 3 and 15 year-old stems, respectively, 34 and 39% produce siderophores; 22 and 15% secrete ammonia; 22 and 21% produce indole-3-acetic acid; 8.7 and 41% solubilize phosphate. Besides, two strains isolated from 15 year-old grapevines showed 1-aminocyclopropane-1-carboxylate deaminase activity. Antifungal activity analysis evidenced that two Bacillus strains possess growth antagonistic effect toward all the tested fungal strains. Therefore, the present study extends our knowledge of the diversity of the endophytic bacteria by providing new insights into the complexity of the grapevine microbiome. PMID:26805617

  15. Biocontrol potential of endophytes harbored in Radula marginata (liverwort) from the New Zealand ecosystem.

    PubMed

    Kusari, Parijat; Kusari, Souvik; Spiteller, Michael; Kayser, Oliver

    2014-10-01

    Radula marginata and Cannabis sativa L. are two phylogenetically unrelated plant species containing structurally similar secondary metabolites like cannabinoids. The major objective of our work was the isolation, identification, biocontrol efficacies, biofilm forming potential and anti-biofilm ability of endophytic microbial community of the liverwort R. marginata, as compared to bacterial endophytic isolates harbored in C. sativa plants. A total of 15 endophytic fungal and 4 endophytic bacterial isolates were identified, including the presence of a bacterial endosymbiont within an endophytic fungal isolate. The endosymbiont was visible only when the fungus containing it was challenged with two phytopathogens Botrytis cinerea and Trichothecium roseum, highlighting a tripartite microbe-microbe interaction and biocontrol potency of endophytes under biotic stress. We also observed sixteen types of endophytic fungal-pathogen and twelve types of endophytic bacterial-pathogen interactions coupled to varying degree of growth inhibitions of either the pathogen or endophyte or both. This showed the magnitude of biocontrol efficacies of endophytes in aiding plant fitness benefits under different media (environmental) conditions. Additionally, it was ecologically noteworthy to find the presence of similar endophytic bacterial genera in both Radula and Cannabis plants, which exhibited similar functional traits like biofilm formation and general anti-biofilm activities. Thus far, our work underlines the biocontrol potency and defensive functional traits (in terms of antagonism and biofilm formation) of endophytes harbored in liverwort R. marginata as compared to the endophytic community of phylogenetically unrelated but phytochemically similar plant C. sativa. PMID:25100187

  16. Genomic and chemical insights into biosurfactant production by the mangrove-derived strain Bacillus safensis CCMA-560.

    PubMed

    Domingos, Daniela Ferreira; de Faria, Andreia Fonseca; de Souza Galaverna, Renan; Eberlin, Marcos Nogueira; Greenfield, Paul; Zucchi, Tiago Domingues; Melo, Itamar Soares; Tran-Dinh, Nai; Midgley, David; de Oliveira, Valéria Maia

    2015-04-01

    Many Bacillus species can produce biosurfactant, although most of the studies on lipopeptide production by this genus have been focused on Bacillus subtilis. Surfactants are broadly used in pharmaceutical, food and petroleum industry, and biological surfactant shows some advantages over the chemical surfactants, such as less toxicity, production from renewable, cheaper feedstocks and development of novel recombinant hyperproducer strains. This study is aimed to unveil the biosurfactant metabolic pathway and chemical composition in Bacillus safensis strain CCMA-560. The whole genome of the CCMA-560 strain was previously sequenced, and with the aid of bioinformatics tools, its biosurfactant metabolic pathway was compared to other pathways of closely related species. Fourier transform infrared (FTIR) and high-resolution TOF mass spectrometry (MS) were used to characterize the biosurfactant molecule. B. safensis CCMA-560 metabolic pathway is similar to other Bacillus species; however, some differences in amino acid incorporation were observed, and chemical analyses corroborated the genetic results. The strain CCMA-560 harbours two genes flanked by srfAC and srfAD not present in other Bacillus spp., which can be involved in the production of the analogue gramicidin. FTIR and MS showed that B. safensis CCMA-560 produces a mixture of at least four lipopeptides with seven amino acids incorporated and a fatty acid chain with 14 carbons, which makes this molecule similar to the biosurfactant of Bacillus pumilus, namely, pumilacidin. This is the first report on the biosurfactant production by B. safensis, encompassing the investigation of the metabolic pathway and chemical characterization of the biosurfactant molecule. PMID:25586584

  17. Diversity of sporadic symbionts and nonsymbiotic endophytic bacteria isolated from nodules of woody, shrub, and food legumes in Ethiopia.

    PubMed

    Aserse, Aregu Amsalu; Räsänen, Leena A; Aseffa, Fassil; Hailemariam, Asfaw; Lindström, Kristina

    2013-12-01

    Fifty-five bacterial isolates were obtained from surface-sterilized nodules of woody and shrub legumes growing in Ethiopia: Crotalaria spp., Indigofera spp., and Erythrina brucei, and the food legumes soybean and common bean. Based on partial 16S rRNA gene sequence analysis, the majority of the isolates were identified as Gram-negative bacteria belonging to the genera Achromobacter, Agrobacterium, Burkholderia, Cronobacter, Enterobacter, Mesorhizobium, Novosphingobium, Pantoea, Pseudomonas, Rahnella, Rhizobium, Serratia, and Variovorax. Seven isolates were Gram-positive bacteria belonging to the genera Bacillus, Paenibacillus, Planomicrobium, and Rhodococcus. Amplified fragment length polymorphism (AFLP) fingerprinting showed that each strain was genetically distinct. According to phylogenetic analysis of recA, glnII, rpoB, and 16S rRNA gene sequences, Rhizobium, Mesorhizobium, and Agrobacterium were further classified into six different genospecies: Agrobacterium spp., Agrobacterium radiobacter, Rhizobium sp., Rhizobium phaseoli, Mesorhizobium sp., and putative new Rhizobium species. The strains from R. phaseoli, Rhizobium sp. IAR30, and Mesorhizobium sp. ERR6 induced nodules on their host plants. The other strains did not form nodules on their original host. Nine endophytic bacterial strains representing seven genera, Agrobacterium, Burkholderia, Paenibacillus, Pantoea, Pseudomonas, Rhizobium, and Serratia, were found to colonize nodules of Crotalaria incana and common bean on co-inoculation with symbiotic rhizobia. Four endophytic Rhizobium and two Agrobacterium strains had identical nifH gene sequences with symbiotic Rhizobium strains, suggesting horizontal gene transfer. Most symbiotic and nonsymbiotic endophytic bacteria showed plant growth-promoting properties in vitro, which indicate their potential role in the promotion of plant growth when colonizing plant roots and the rhizosphere. PMID:24196581

  18. Rare actinomycetes Nocardia caishijiensis and Pseudonocardia carboxydivorans as endophytes, their bioactivity and metabolites evaluation.

    PubMed

    Tanvir, Rabia; Sajid, Imran; Hasnain, Shahida; Kulik, Andreas; Grond, Stephanie

    2016-04-01

    Two strains identified as Nocardia caishijiensis (SORS 64b) and Pseudonocardia carboxydivorans (AGLS 2) were isolated as endophytes from Sonchus oleraceus and Ageratum conyzoides respectively. The analysis of their extracts revealed them to be strongly bioactive. The N. caishijiensis extract gave an LC50 of 570μg/ml(-1) in the brine shrimp cytotoxicity assay and an EC50 of 0.552μg/ml(-1) in the DPPH antioxidant assay. Antimicrobial activity was observed against Methicillin resistant Staphlococcus aureus (MRSA) and Escherichia coli ATCC 25922 (14mm), Klebsiella pneumoniae ATCC 706003 (13mm), S. aureus ATCC 25923 (11mm) and Candida tropicalis (20mm). For the extract of P. carboxydivorans the EC50 was 0.670μg/ml(-1) and it was observed to be more bioactive against Bacillus subtilis DSM 10 ATCC 6051 (21mm), C. tropicalis (20mm), S. aureus ATCC 25923 (17mm), MRSA (17mm), E. coli K12 (W1130) (16mm) and Chlorella vulgaris (10mm). The genotoxicity testing revealed a 20mm zone of inhibition against the polA mutant strain E. coli K-12 AB 3027 suggesting damage to the DNA and polA genes. The TLC and bioautography screening revealed a diversity of active bands of medium polar and nonpolar compounds. Metabolite analysis by HPLC-DAD via UV/vis spectral screening suggested the possibility of stenothricin and bagremycin A in the mycelium extract of N. caishijiensis respectively. In the broth and mycelium extract of P. carboxydivorans borrelidin was suggested along with α-pyrone. The HPLC-MS revealed bioactive long chained amide derivatives such as 7-Octadecenamide, 9, 12 octadecandienamide. This study reports the rare actinomycetes N. caishijiensis and P. carboxydivorans as endophytes and evaluates their bioactive metabolites. PMID:26946375

  19. Endophytic actinomycetes from Azadirachta indica A. Juss.: isolation, diversity, and anti-microbial activity.

    PubMed

    Verma, Vijay C; Gond, Surendra K; Kumar, Anuj; Mishra, Ashish; Kharwar, Ravindra N; Gange, Alan C

    2009-05-01

    Endophytic actinomycetes from Azadirachta indica A. Juss. were screened and evaluated for their anti-microbial activity against an array of pathogenic fungi and bacteria. A total of 55 separate isolates were obtained from 20 plants, and 60% of these showed inhibitory activity against one or more pathogenic fungi and bacteria. Actinomycetes were most commonly recovered from roots (54.5% of all isolates), followed by stems (23.6%), and leaves (21.8%). The dominant genus was Streptomyces (49.09% of all isolates), while Streptosporangium (14.5%), Microbispora (10.9%), Streptoverticillium (5.5%), Sacchromonospora sp. (5.5%), and Nocardia (3.6%) were also recovered. Streptomyces isolates AzR 006, 011, and 031 (all from roots) had acute activity against Pseudomonas fluorescens, while AzR027, 032, and 051 (also all from roots) showed activity against Escherichia coli. Meanwhile, an isolate of Nocardia sp. from leaves (AzL025) showed antagonism against Bacillus subtilis. Overall, 32 of the 55 were found to have broad spectrum significant antimicrobial activity, while about 4% of them showed strong and acute inhibition to pathogenic fungi and bacteria. Isolates of Streptomyces AzR031, 008, and 047, Nocardia sp. AzL025, and Streptosporangium sp. AzR 021 and 048 are of particular interest because they showed significant antagonistic activity against root pathogens, including Pythium and Phytophthora sp. Thus, many of the isolates recovered from A. indica in this study may be used in developing potential bio-control agents against a range of pathogenic fungi and bacteria and in the production of novel natural antimicrobial compounds. These results not only further our understanding of plant-microbe interactions but also indicate that there is an untapped resource of endophytic microorganisms that could be exploited in the biotechnological, medicinal, and agricultural industries. PMID:18853084

  20. Biodegradation of Polyester Polyurethane by Endophytic Fungi?

    PubMed Central

    Russell, Jonathan R.; Huang, Jeffrey; Anand, Pria; Kucera, Kaury; Sandoval, Amanda G.; Dantzler, Kathleen W.; Hickman, DaShawn; Jee, Justin; Kimovec, Farrah M.; Koppstein, David; Marks, Daniel H.; Mittermiller, Paul A.; Núñez, Salvador Joel; Santiago, Marina; Townes, Maria A.; Vishnevetsky, Michael; Williams, Neely E.; Vargas, Mario Percy Núñez; Boulanger, Lori-Ann; Bascom-Slack, Carol; Strobel, Scott A.

    2011-01-01

    Bioremediation is an important approach to waste reduction that relies on biological processes to break down a variety of pollutants. This is made possible by the vast metabolic diversity of the microbial world. To explore this diversity for the breakdown of plastic, we screened several dozen endophytic fungi for their ability to degrade the synthetic polymer polyester polyurethane (PUR). Several organisms demonstrated the ability to efficiently degrade PUR in both solid and liquid suspensions. Particularly robust activity was observed among several isolates in the genus Pestalotiopsis, although it was not a universal feature of this genus. Two Pestalotiopsis microspora isolates were uniquely able to grow on PUR as the sole carbon source under both aerobic and anaerobic conditions. Molecular characterization of this activity suggests that a serine hydrolase is responsible for degradation of PUR. The broad distribution of activity observed and the unprecedented case of anaerobic growth using PUR as the sole carbon source suggest that endophytes are a promising source of biodiversity from which to screen for metabolic properties useful for bioremediation. PMID:21764951

  1. Biodegradation of polyester polyurethane by endophytic fungi.

    PubMed

    Russell, Jonathan R; Huang, Jeffrey; Anand, Pria; Kucera, Kaury; Sandoval, Amanda G; Dantzler, Kathleen W; Hickman, DaShawn; Jee, Justin; Kimovec, Farrah M; Koppstein, David; Marks, Daniel H; Mittermiller, Paul A; Núñez, Salvador Joel; Santiago, Marina; Townes, Maria A; Vishnevetsky, Michael; Williams, Neely E; Vargas, Mario Percy Núñez; Boulanger, Lori-Ann; Bascom-Slack, Carol; Strobel, Scott A

    2011-09-01

    Bioremediation is an important approach to waste reduction that relies on biological processes to break down a variety of pollutants. This is made possible by the vast metabolic diversity of the microbial world. To explore this diversity for the breakdown of plastic, we screened several dozen endophytic fungi for their ability to degrade the synthetic polymer polyester polyurethane (PUR). Several organisms demonstrated the ability to efficiently degrade PUR in both solid and liquid suspensions. Particularly robust activity was observed among several isolates in the genus Pestalotiopsis, although it was not a universal feature of this genus. Two Pestalotiopsis microspora isolates were uniquely able to grow on PUR as the sole carbon source under both aerobic and anaerobic conditions. Molecular characterization of this activity suggests that a serine hydrolase is responsible for degradation of PUR. The broad distribution of activity observed and the unprecedented case of anaerobic growth using PUR as the sole carbon source suggest that endophytes are a promising source of biodiversity from which to screen for metabolic properties useful for bioremediation. PMID:21764951

  2. Fungal endophyte metabolism and allelopathic interactions with host plants

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The maize endophyte Fusarium verticillioides produces the fumonisin mycotoxins, which are of significant concern for their animal toxicity caused by inhibition of ceramide synthase and disruption of sphingolipid metabolism. Fumonisin-producing strains associated with maize cause leaf lesions, develo...

  3. Chemical Assessment and Antimicrobial and Antioxidant Activities of Endophytic Fungi Extracts Isolated from Costus spiralis (Jacq.) Roscoe (Costaceae)

    PubMed Central

    Marson Ascêncio, Poliana Guerino; Ascêncio, Sérgio Donizeti; Aguiar, Aline Aires; Fiorini, Adriana; Pimenta, Raphael Sanzio

    2014-01-01

    Costus spiralis (Costaceae) is a species native to the Amazon region and is used in traditional medicine. The endophytic fungi used in this study were obtained from leaves of this plant. 13 strains were selected to obtain hydroethanolic extracts and were submitted to hydroalcoholic extraction and evaluated for antioxidant activity by DPPH (2,2-difenil-1-picrilhidrazil) and FRAP (ferric reducing antioxidant power), and all of the fungi had positive results. The antimicrobial action of crude extracts had a good range of activities. All extracts had inhibitory activities against the yeasts of Candida albicans and C. parapsilosis, with 125 to 500 μg/mL MIC. Eight extracts had antimicrobial activities against Bacillus subtilis (MIC from 62.4 to 125 μg/mL), 5 against Pseudomonas aeruginosa (MIC from 125 to 500 μg/mL), 2 against Salmonella enterica (MIC from 125 to 62.5 μg/mL), and 2 against Enterococcus faecalis (MIC from 500 to 125 μg/mL). The presence of secondary metabolites, including coumarins, was observed during chemical evaluation by thin layer chromatography. Total phenol content was estimated, and a strong positive correlation to antioxidant activity was observed, according to its Pearson coefficient. This is the first report of the bioactive potential of endophytic fungi isolated from the Costaceae family in Brazilian ecosystems. PMID:25587339

  4. Diversity of endophytic bacteria associated with nodules of two indigenous legumes at different altitudes of the Qilian Mountains in China.

    PubMed

    Xu, Lin; Zhang, Yong; Wang, Li; Chen, Weimin; Wei, Gehong

    2014-09-01

    A total of 201 endophytic root nodule-associated bacteria collected from two legumes indigenous to different Qilian Mountain altitudes (Hexi Corridor) were characterized through 16S rDNA polymerase chain reaction (PCR)-restriction fragment length polymorphism, 16S rRNA gene sequence analysis, and enterobacterial repetitive intergenic consensus-PCR clustering. The isolates phylogenetically belonged to 35 species in the Phyllobacterium, Ensifer, Rhizobium, Microvirga, Sphingomonas, Paracoccus, Mycobacterium, Paenibacillus, Cohnella, Sporosarcina, Bacillus, Staphylococcus, Brevibacterium, Xenophilus, Erwinia, Leclercia, Acinetobacter, and Pseudomonas genera. Phylogenetic nodA sequence analysis showed higher similarity to Sinorhizobium meliloti with strains related to the Rhizobium, Sinorhizobium, and Acinetobacter genera. Sequence analysis of the nifH gene revealed that the strains belonging to Xenophilus, Acinetobacter, Phyllobacterium, and Rhizobium had genes similar to those of Mesorhizobium and Sinorhizobium. The results indicated that horizontal gene transfer could have occurred between rhizobia and non-rhizobial endophytes. Canonical correspondence analysis revealed that altitude and host plant species contributed more to the bacterial endosymbiont separation than other ecological factors. This study provided valuable information on the interactions between symbiotic bacteria, non-symbiotic bacteria and their habitats, and thus provided knowledge on their genetic diversity and ecology. PMID:24985194

  5. Diversity of endophytic bacteria within nodules of the Sphaerophysa salsula in different regions of Loess Plateau in China.

    PubMed

    Deng, Zhen Shan; Zhao, Long Fei; Kong, Zhao Yu; Yang, Wen Quan; Lindström, Kristina; Wang, En Tao; Wei, Ge Hong

    2011-06-01

    A total of 115 endophytic bacteria were isolated from root nodules of the wild legume Sphaerophysa salsula grown in two ecological regions of Loess Plateau in China. The genetic diversity and phylogeny of the strains were revealed by restriction fragment length polymorphism and sequencing of 16S rRNA gene and enterobacterial repetitive intergenic consensus-PCR. Their symbiotic capacity was checked by nodulation tests and analysis of nifH gene sequence. This is the first systematic study on endophytic bacteria associated with S. salsula root nodules. Fifty of the strains found were symbiotic bacteria belonging to eight putative species in the genera Mesorhizobium, Rhizobium and Sinorhizobium, harboring similar nifH genes; Mesorhizobium gobiense was the main group and 65 strains were nonsymbiotic bacteria related to 17 species in the genera Paracoccus, Sphingomonas, Inquilinus, Pseudomonas, Serratia, Mycobacterium, Nocardia, Streptomyces, Paenibacillus, Brevibacillus, Staphylococcus, Lysinibacillus and Bacillus, which were universally coexistent with symbiotic bacteria in the nodules. Differing from other similar studies, the present study is the first time that symbiotic and nonsymbiotic bacteria have been simultaneously isolated from the same root nodules, offering the possibility to accurately reveal the correlation between these two kinds of bacteria. These results provide valuable information about the interactions among the symbiotic bacteria, nonsymbiotic bacteria and their habitats. PMID:21303396

  6. Ergot alkaloids produced by endophytic fungi of the genus Epichloë.

    PubMed

    Guerre, Philippe

    2015-03-01

    The development of fungal endophytes of the genus Epichloë in grasses results in the production of different groups of alkaloids, whose mechanism and biological spectrum of toxicity can differ considerably. Ergot alkaloids, when present in endophyte-infected tall fescue, are responsible for "fescue toxicosis" in livestock, whereas indole-diterpene alkaloids, when present in endophyte-infected ryegrass, are responsible for "ryegrass staggers". In contrast, peramine and loline alkaloids are deterrent and/or toxic to insects. Other toxic effects in livestock associated with the consumption of endophyte-infected grass that contain ergot alkaloids include the "sleepy grass" and "drunken horse grass" diseases. Although ergovaline is the main ergopeptine alkaloid produced in endophyte-infected tall fescue and is recognized as responsible for fescue toxicosis, a number of questions still exist concerning the profile of alkaloid production in tall fescue and the worldwide distribution of tall fescue toxicosis. The purpose of this review is to present ergot alkaloids produced in endophyte-infected grass, the factors of variation of their level in plants, and the diseases observed in the mammalian species as relate to the profiles of alkaloid production. In the final section, interactions between ergot alkaloids and drug-metabolizing enzymes are presented as mechanisms that could contribute to toxicity. PMID:25756954

  7. Exploring Arabidopsis thaliana Root Endophytes via Single-Cell Genomics

    SciTech Connect

    Lundberg, Derek; Woyke, Tanja; Tringe, Susannah; Dangl, Jeff

    2014-03-19

    Land plants grow in association with microbial communities both on their surfaces and inside the plant (endophytes). The relationships between microbes and their host can vary from pathogenic to mutualistic. Colonization of the endophyte compartment occurs in the presence of a sophisticated plant immune system, implying finely tuned discrimination of pathogens from mutualists and commensals. Despite the importance of the microbiome to the plant, relatively little is known about the specific interactions between plants and microbes, especially in the case of endophytes. The vast majority of microbes have not been grown in the lab, and thus one of the few ways of studying them is by examining their DNA. Although metagenomics is a powerful tool for examining microbial communities, its application to endophyte samples is technically difficult due to the presence of large amounts of host plant DNA in the sample. One method to address these difficulties is single-cell genomics where a single microbial cell is isolated from a sample, lysed, and its genome amplified by multiple displacement amplification (MDA) to produce enough DNA for genome sequencing. This produces a single-cell amplified genome (SAG). We have applied this technology to study the endophytic microbes in Arabidopsis thaliana roots. Extensive 16S gene profiling of the microbial communities in the roots of multiple inbred A. thaliana strains has identified 164 OTUs as being significantly enriched in all the root endophyte samples compared to their presence in bulk soil.

  8. Ungulate saliva inhibits a grass–endophyte mutualism

    PubMed Central

    Tanentzap, Andrew J.; Vicari, Mark; Bazely, Dawn R.

    2014-01-01

    Fungal endophytes modify plant–herbivore interactions by producing toxic alkaloids that deter herbivory. However, studies have neglected the direct effects herbivores may have on endophytes. Antifungal properties and signalling effectors in herbivore saliva suggest that evolutionary pressures may select for animals that mitigate the effects of endophyte-produced alkaloids. Here, we tested whether saliva of moose (Alces alces) and European reindeer (Rangifer tarandus) reduced hyphal elongation and production of ergot alkaloids by the foliar endophyte Epichloë festucae associated with the globally distributed red fescue Festuca rubra. Both moose and reindeer saliva reduced the growth of isolated endophyte hyphae when compared with a treatment of distilled water. Induction of the highly toxic alkaloid ergovaline was also inhibited in plants from the core of F. rubra's distribution when treated with moose saliva following simulated grazing. In genotypes from the southern limit of the species' distribution, ergovaline was constitutively expressed, as predicted where growth is environmentally limited. Our results now present the first evidence, to our knowledge, that ungulate saliva can combat plant defences produced by a grass–endophyte mutualism. PMID:25055816

  9. Ungulate saliva inhibits a grass-endophyte mutualism.

    PubMed

    Tanentzap, Andrew J; Vicari, Mark; Bazely, Dawn R

    2014-07-01

    Fungal endophytes modify plant-herbivore interactions by producing toxic alkaloids that deter herbivory. However, studies have neglected the direct effects herbivores may have on endophytes. Antifungal properties and signalling effectors in herbivore saliva suggest that evolutionary pressures may select for animals that mitigate the effects of endophyte-produced alkaloids. Here, we tested whether saliva of moose (Alces alces) and European reindeer (Rangifer tarandus) reduced hyphal elongation and production of ergot alkaloids by the foliar endophyte Epichloë festucae associated with the globally distributed red fescue Festuca rubra. Both moose and reindeer saliva reduced the growth of isolated endophyte hyphae when compared with a treatment of distilled water. Induction of the highly toxic alkaloid ergovaline was also inhibited in plants from the core of F. rubra's distribution when treated with moose saliva following simulated grazing. In genotypes from the southern limit of the species' distribution, ergovaline was constitutively expressed, as predicted where growth is environmentally limited. Our results now present the first evidence, to our knowledge, that ungulate saliva can combat plant defences produced by a grass-endophyte mutualism. PMID:25055816

  10. Symbiotic grasses: A review of basic biology of forage grass fungal endophytes

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The fungal endophytes associated with grasses are the fundamental reason for the basic successes of several pasture grasses, notable tall fescues, and perennial ryegrass. Tall fescue and perennial ryegrass fungal endophytes, Neotyphodium coenophialum and N. lolii, respectively, and their relatives ...

  11. Asexual endophytes in a native grass: Tradeoffs in mortality, growth, reproduction, and alkaloid production

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Neotyphodium endophytes are asexual, seed-borne fungal symbionts that are thought to interact mutualistically with their grass hosts. Benefits include increased growth, reproduction, and resistance of herbivores via endophytic alkaloids. Although these benefits are well established in infected int...

  12. Beauveria bassiana and Metarhizium anisopliae endophytically colonize cassava roots following soil drench inoculation

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The fungal entomopathogens Beauveria bassiana and Metarhizium anisopliae were investigated to determine if endophytic colonization could be achieved in cassava. An inoculation method based on drenching the soil around cassava stems using conidial suspensions resulted in endophytic colonization of ca...

  13. ENHANCEMENT OF CHILE BIOMAS, FRUIT QUALITY, AND DROUGHT STRESS TOLERANCE BY INOCULATION WITH NATIVE PLANT ENDOPHYTES

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Recognition of the important contributions endophytes make to plant stress tolerance, defense, and production has grown exponentially with improvements in detection of unculturable microorganisms in plant tissues. Recently, the ability to manipulate host-endophyte combinations has provided a powerf...

  14. Fungal endophyte infection increases carbon sequestration potential of southeastern USA tall fescue stands

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Tall fescue (Schedonorus arundinaceous (Schreb.)) is often infected with a common toxic fungal endophyte (Neotyphodium coenophialum) capable of producing alkaloids that affect grazing animal health, insect herbivory, plant production, and litter decomposition. The strength of these endophyte-associa...

  15. Preferential Association of Endophytic Bradyrhizobia with Different Rice Cultivars and Its Implications for Rice Endophyte Evolution

    PubMed Central

    Piromyou, Pongdet; Greetatorn, Teerana; Teamtisong, Kamonluck; Okubo, Takashi; Shinoda, Ryo; Nuntakij, Achara; Tittabutr, Panlada; Boonkerd, Nantakorn

    2015-01-01

    Plant colonization by bradyrhizobia is found not only in leguminous plants but also in nonleguminous species such as rice. To understand the evolution of the endophytic symbiosis of bradyrhizobia, the effect of the ecosystems of rice plantations on their associations was investigated. Samples were collected from various rice (Oryza sativa) tissues and crop rotational systems. The rice endophytic bradyrhizobia were isolated on the basis of oligotrophic properties, selective medium, and nodulation on siratro (Macroptilium atropurpureum). Six bradyrhizobial strains were obtained exclusively from rice grown in a crop rotational system. The isolates were separated into photosynthetic bradyrhizobia (PB) and nonphotosynthetic bradyrhizobia (non-PB). Thai bradyrhizobial strains promoted rice growth of Thai rice cultivars better than the Japanese bradyrhizobial strains. This implies that the rice cultivars possess characteristics that govern rice-bacterium associations. To examine whether leguminous plants in a rice plantation system support the persistence of rice endophytic bradyrhizobia, isolates were tested for legume nodulation. All PB strains formed symbioses with Aeschynomene indica and Aeschynomene evenia. On the other hand, non-PB strains were able to nodulate Aeschynomene americana, Vigna radiata, and M. atropurpureum but unable to nodulate either A. indica or A. evenia. Interestingly, the nodABC genes of all of these bradyrhizobial strains seem to exhibit low levels of similarity to those of Bradyrhizobium diazoefficiens USDA110 and Bradyrhizobium sp. strain ORS285. From these results, we discuss the evolution of the plant-bradyrhizobium association, including nonlegumes, in terms of photosynthetic lifestyle and nod-independent interactions. PMID:25710371

  16. Microarray and Real-Time PCR Comparison of Endophyte-Infected and Endophyte-Free Tall Fescue Gene Expression

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Many grasses have mutualistic symbioses with fungi of the family Clavicipitaceae. Tall fescue [Schedonorus arundinaceus (Schreb.) Dumont. = Festuca arundinacea (Schreb.)] can harbor the obligate endophyte, Neotyphodium coenophialum, that is asexually propagated and transmitted via host seeds. To d...

  17. Altitude and forest edges influence the density and distribution of pygmy tarsiers (Tarsius pumilus).

    PubMed

    Grow, Nanda; Gursky, Sharon; Duma, Yulius

    2013-05-01

    In this study, we examine how high-altitude ecology and anthropogenic edges relate to the density and distribution of pygmy tarsiers. Pygmy tarsiers (Tarsius pumilus) are extremely small-bodied primates (55 g) that are endemic to high-altitude forest and exhibit several differences from lowland Sulawesian tarsier species. From June to September 2010 and January to March 2012, we conducted a population census of pygmy tarsiers across multiple altitudes. Sampling took place within a 1.2 km(2) area encompassing altitudes of 2,000-2,300 m a.s.l. on Mt. Rore Katimbu in Lore Lindu National Park, central Sulawesi, Indonesia. We observed 22 individuals, with an estimated population density of 92 individuals per 100 ha. These results indicate that pygmy tarsiers live at a lower density than lowland Sulawesian tarsier species. Lower density was associated with decreased resources at higher altitudes, including decreased tree size, tree density, and insect biomass. Within the sample area, we found pygmy tarsiers in only 8 of 24 (33%) quadrats, suggesting a nonrandom distribution that probably overinflated this population density estimate. Pygmy tarsiers exhibited a clumped distribution near anthropogenic edges that were associated with increased insect abundance and biomass. Airborne insects were more abundant along forest edges than within the forest interior, and pygmy tarsiers were observed to forage along edges where there was a higher abundance of Lepidoptera and Orthoptera. Tarsiers may mitigate the decreased availability of insects at high altitudes by adjusting their ranging patterns to remain near forest edges. PMID:23325720

  18. Bioactivity of Fungal Endophytes as a Function of Endophyte Taxonomy and the Taxonomy and Distribution of Their Host Plants

    PubMed Central

    Higginbotham, Sarah J.; Arnold, A. Elizabeth; Ibañez, Alicia; Spadafora, Carmenza; Coley, Phyllis D.; Kursar, Thomas A.

    2013-01-01

    Fungal endophytes – fungi that grow within plant tissues without causing immediate signs of disease – are abundant and diverse producers of bioactive secondary metabolites. Endophytes associated with leaves of tropical plants are an especially exciting and relatively untapped source of novel compounds. However, one major challenge in drug discovery lies in developing strategies to efficiently recover highly bioactive strains. As part of a 15-year drug discovery project, foliar endophytes were isolated from 3198 plant samples (51 orders, 105 families and at least 232 genera of angiosperms and ferns) collected in nine geographically distinct regions of Panama. Extracts from culture supernatants of >2700 isolates were tested for bioactivity (in vitro percent inhibition of growth, % IG) against a human breast cancer cell line (MCF-7) and the causative agents of malaria, leishmaniasis, and Chagas' disease. Overall, 32.7% of endophyte isolates were highly active in at least one bioassay, including representatives of diverse fungal lineages, host lineages, and collection sites. Up to 17% of isolates tested per assay were highly active. Most bioactive strains were active in only one assay. Fungal lineages differed in the incidence and degree of bioactivity, as did fungi from particular plant taxa, and greater bioactivity was observed in endophytes isolated from plants in cloud forests vs. lowland forests. Our results suggest that using host taxonomy and forest type to tailor plant collections, and selecting endophytes from specific orders or families for cultivation, will markedly increase the efficiency and efficacy of discovering bioactive metabolites for particular pharmaceutical targets. PMID:24066037

  19. Bacillus odysseyi isolate

    NASA Technical Reports Server (NTRS)

    Venkateswaran, Kasthuri (Inventor); La Duc, Myron Thomas (Inventor)

    2007-01-01

    The present invention relates to discovery and isolation of a biologically pure culture of a Bacillus odysseyi isolate with high adherence and sterilization resistant properties. B. odysseyi is a round spore forming Bacillus species that produces an exosporium. This novel species has been characterized on the basis of phenotypic traits, 16S rDNA sequence analysis and DNA-DNA hybridization. According to the results of these analyses, this strain belongs to the genus Bacillus and the type strain is 34hs-1.sup.T (=ATCC PTA-4993.sup.T=NRRL B-30641.sup.T=NBRC 100172.sup.T). The GenBank accession number for the 16S rDNA sequence of strain 34hs-1.sup.T is AF526913.

  20. Endophytes as in vitro production platforms of high value plant secondary metabolites.

    PubMed

    Venugopalan, Aarthi; Srivastava, Smita

    2015-11-01

    Many reports have been published on bioprospecting of endophytic fungi capable of producing high value bioactive molecules like, paclitaxel, vincristine, vinblastine, camptothecin and podophyllotoxin. However, commercial exploitation of endophytes for high value-low volume plant secondary metabolites remains elusive due to widely reported genomic instability of endophytes in the axenic culture. While most of the endophyte research focuses on screening endophytes for novel or existing high value biomolecules, very few reports seek to explore the possible mechanisms of production of host-plant associated or novel secondary metabolites in these organisms. With an overview of host-endophyte relationship and its possible impact on the secondary metabolite production potential of endophytes, the review highlights the evidence reported for and against the presence of host-independent biosynthetic machinery in endophytes. The review aims to address the question, why should and how can endophytes be exploited for large scale in vitro production of high value phytochemicals? In this regard, various bioprocess optimization strategies that have been applied to sustain and enhance the product yield from the endophytes have also been described in detail. Further, techniques like mixed fermentation/co-cultivation and use of epigenetic modifiers have also been discussed as potential strategies to activate cryptic gene clusters in endophytes, thereby aiding in novel metabolite discovery and overcoming the limitations associated with axenic culture of endophytes. PMID:26225453

  1. Tall fescue management: Pasture and cattle responses to endophyte and fertilization

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Yearling heifers grazing tall fescue pastures had greatest performance in winter and spring on endophyte-free and novel endophyte associations, because of high forage quality and lack of ergot alkaloids produced by a common “wild” tall fescue-endophyte association. Pasture and cattle responses were...

  2. Alkaloid variation among epichloid endophytes of sleepygrass (Achnatherum robustum) and consequences for resistance to insect herbivores.

    PubMed

    Shymanovich, Tatsiana; Saari, Susanna; Lovin, Mary E; Jarmusch, Alan K; Jarmusch, Scott A; Musso, Ashleigh M; Charlton, Nikki D; Young, Carolyn A; Cech, Nadja B; Faeth, Stanley H

    2015-01-01

    Epichloid endophytes are well known symbionts of many cool-season grasses that may alleviate environmental stresses for their hosts. For example, endophytes produce alkaloid compounds that may be toxic to invertebrate or vertebrate herbivores. Achnatherum robustum, commonly called sleepygrass, was aptly named due to the presence of an endophyte that causes toxic effects to livestock and wildlife. Variation in alkaloid production observed in two A. robustum populations located near Weed and Cloudcroft in the Lincoln National Forest, New Mexico, suggests two different endophyte species are present in these populations. Genetic analyses of endophyte-infected samples revealed major differences in the endophyte alkaloid genetic profiles from the two populations, which were supported with chemical analyses. The endophyte present in the Weed population was shown to produce chanoclavine I, paspaline, and terpendoles, so thus resembles the previously described Epichloë funkii. The endophyte present in the Cloudcroft population produces chanoclavineI, ergonovine, lysergic acid amide, and paspaline, and is an undescribed endophyte species. We observed very low survival rates for aphids feeding on plants infected with the Cloudcroft endophyte, while aphid survival was better on endophyte infected plants in the Weed population. This observation led to the hypothesis that the alkaloid ergonovine is responsible for aphid mortality. Direct testing of aphid survival on oat leaves supplemented with ergonovine provided supporting evidence for this hypothesis. The results of this study suggest that alkaloids produced by the Cloudcroft endophyte, specifically ergonovine, have insecticidal properties. PMID:25501262

  3. Tall fescue endophyte effects on tolerance to water-deficit stress

    Technology Transfer Automated Retrieval System (TEKTRAN)

    To understand more about the enhanced drought tolerance conferred by the endophyte Neotyphodium coenophialum in tall fescue, we analyzed the effect of the endophyte on genetically identical tall fescue clones with (E+) and without the endophyte (E-), by generating E- plants through fungicide trea...

  4. Increased milk production by Holstein cows consuming endophyte-infected fescue seed during the dry period.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Ergot alkaloids in endophyte-infected grasses inhibit prolactin (PRL) secretion and may reduce milk production of cows consuming endophyte-infected grasses. We hypothesized that consumption of endophyte-infected fescue during the dry period inhibits mammary differentiation and subsequent milk produ...

  5. Host suitability of an endophyte-friendly tall fescue grass to Mesocriconema xenoplax and Pratylenchus vulnus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Tall fescue grass cultivars with or without endophytes were evaluated for their susceptibility to Mesocriconema xenoplax and Pratylenchus vulnus in the greenhouse. Tall fescue cultivars evaluated included, i) wild-type Jesup (E+, ergot-producing endophyte present), ii) endophyte-free Jesup (E-, no ...

  6. Leaf endophyte load and fungal garden development in leaf-cutting ants

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Previous work has shown that leaf-cutting ants prefer to cut leaf material that is relatively low in fungal endophyte content. Such a preference suggests that fungal endophytes exact a cost on the ants or on the development of their colonies. We hypothesized that endophytes may play a role in thei...

  7. Endophytic fungi: a reservoir of antibacterials

    PubMed Central

    Deshmukh, Sunil K.; Verekar, Shilpa A.; Bhave, Sarita V.

    2015-01-01

    Multidrug drug resistant bacteria are becoming increasingly problematic particularly in the under developed countries of the world. The most important microorganisms that have seen a geometric rise in numbers are Methicillin resistant Staphylococcus aureus, Vancomycin resistant Enterococcus faecium, Penicillin resistant Streptococcus pneumonia and multiple drug resistant tubercule bacteria to name a just few. New drug scaffolds are essential to tackle this every increasing problem. These scaffolds can be sourced from nature itself. Endophytic fungi are an important reservoir of therapeutically active compounds. This review attempts to present some data relevant to the problem. New, very specific and effective antibiotics are needed but also at an affordable price! A Herculean task for researchers all over the world! In the Asian subcontinent indigenous therapeutics that has been practiced over the centuries such as Ayurveda have been effective as “handed down data” in family generations. May need a second, third and more “in-depth investigations?” PMID:25620957

  8. A novel cephalosporin deacetylating acetyl xylan esterase from Bacillus subtilis with high activity toward cephalosporin C and 7-aminocephalosporanic acid.

    PubMed

    Tian, Qianqian; Song, Ping; Jiang, Ling; Li, Shuang; Huang, He

    2014-03-01

    A cephalosporin deacetylating acetyl xylan esterase was cloned from the genomic DNA of Bacillus subtilis CICC 20034 and functionally expressed in Escherichia coli. Its gene contained an open reading frame of 957 bp encoding 318 amino acids with a calculated mass of 35,607 Da, and it displayed significant identity to acetyl xylan esterases from Bacillus sp. 916, B. subtilis 168, and Bacillus pumilus Cect5072. The enzyme was a native homohexamer but a trimer under the condition of 1% sodium dodecyl sulfate (SDS); both forms were active and could transit to each other by incubating in or removing SDS. The enzyme belongs to carbohydrate esterase family 7 and had a double specificity on both the acetylated oligosaccharide and cephalosporin C (CPC) and 7-aminocephalosporanic acid (7-ACA). The activity of this purified enzyme toward CPC and 7-ACA was highest among all the acetyl xylan esterase from CE family 7, which were 484 and 888 U/mg, respectively, and endowed itself with great industrial interest on semi-synthetic ?-lactam antibiotics. The optimum pH of the purified enzyme was 8.0, and the optimum temperature was 50 °C, and the enzyme had high thermal stability, broad range of pH tolerance, and extremely organic solvent tolerance. PMID:23828600

  9. Isolation and identification of a lipase producing Bacillus sp. from soil.

    PubMed

    Heravi, Kambiz Morabbi; Eftekhar, Fereshteh; Yakhchali, Bagher; Tabandeh, Fatemeh

    2008-03-01

    Lipase production in an indigenous lipolytic Bacillus sp. was detected in media containing Tributyrin-Tween 80 and Rhodamine B-Olive oil. The statistical Taguchi model was used to predict the optimum experimental conditions for bacterial growth and lipase production. Partial optimization was carried out for selection of salt base, oil, glucose, NH4Cl and yeast extract concentrations, inoculum density, pH and agitation. Maximum lipase activity was detected in the cell free supernatants of cultures grown in a medium containing 10 g L(-1) yeast extract, 15 g L(-1) NH4Cl, 3 g L(-1) K2HPO4, 1 g L(-1) KH2PO4, 0.1 g L(-1) MgSO4 x 7H2O, 2 g L(-1) glucose, 0.6 mM MgCl2 and 15 ml L(-1) olive oil, pH 8.5 at 30 degrees C for 24 h and low agitation. The amount oflipase produced in the designed medium was in agreement with the predicted values by the statistical method. 16S rRNA cloning and sequencing identified the test organism as Bacillus pumilus. PMID:18819570

  10. Identification of Bacillus Probiotics Isolated from Soil Rhizosphere Using 16S rRNA, recA, rpoB Gene Sequencing and RAPD-PCR.

    PubMed

    Mohkam, Milad; Nezafat, Navid; Berenjian, Aydin; Mobasher, Mohammad Ali; Ghasemi, Younes

    2016-03-01

    Some Bacillus species, especially Bacillus subtilis and Bacillus pumilus groups, have highly similar 16S rRNA gene sequences, which are hard to identify based on 16S rDNA sequence analysis. To conquer this drawback, rpoB, recA sequence analysis along with randomly amplified polymorphic (RAPD) fingerprinting was examined as an alternative method for differentiating Bacillus species. The 16S rRNA, rpoB and recA genes were amplified via a polymerase chain reaction using their specific primers. The resulted PCR amplicons were sequenced, and phylogenetic analysis was employed by MEGA 6 software. Identification based on 16S rRNA gene sequencing was underpinned by rpoB and recA gene sequencing as well as RAPD-PCR technique. Subsequently, concatenation and phylogenetic analysis showed that extent of diversity and similarity were better obtained by rpoB and recA primers, which are also reinforced by RAPD-PCR methods. However, in one case, these approaches failed to identify one isolate, which in combination with the phenotypical method offsets this issue. Overall, RAPD fingerprinting, rpoB and recA along with concatenated genes sequence analysis discriminated closely related Bacillus species, which highlights the significance of the multigenic method in more precisely distinguishing Bacillus strains. This research emphasizes the benefit of RAPD fingerprinting, rpoB and recA sequence analysis superior to 16S rRNA gene sequence analysis for suitable and effective identification of Bacillus species as recommended for probiotic products. PMID:26898909

  11. Vertical transmission of fungal endophytes is widespread in forbs

    PubMed Central

    Hodgson, Susan; Cates, Catherine; Hodgson, Joshua; Morley, Neil J; Sutton, Brian C; Gange, Alan C

    2014-01-01

    To date, it has been thought that endophytic fungi in forbs infect the leaves of their hosts most commonly by air-borne spores (termed “horizontal transmission”). Here, we show that vertical transmission from mother plant to offspring, via seeds, occurs in six forb species (Centaurea cyanus, C. nigra,Papaver rhoeas,Plantago lanceolata,Rumex acetosa, and Senecio vulgaris), suggesting that this may be a widespread phenomenon. Mature seeds were collected from field-grown plants and endophytes isolated from these, and from subsequent cotyledons and true leaves of seedlings, grown in sterile conditions. Most seeds contain one species of fungus, although the identity of the endophyte differs between plant species. Strong evidence for vertical transmission was found for two endophyte species, Alternaria alternata and Cladosporium sphaerospermum. These fungi were recovered from within seeds, cotyledons, and true leaves, although the plant species they were associated with differed. Vertical transmission appears to be an imperfect process, and germination seems to present a bottleneck for fungal growth. We also found that A. alternata and C. sphaerospermum occur on, and within pollen grains, showing that endophyte transmission can be both within and between plant generations. Fungal growth with the pollen tube is likely to be the way in which endophytes enter the developing seed. The fact that true vertical transmission seems common suggests a more mutualistic association between these fungi and their hosts than has previously been thought, and possession of endophytes by seedling plants could have far-reaching ecological consequences. Seedlings may have different growth rates and be better protected against herbivores and pathogens, dependent on the fungi that were present in the mother plant. This would represent a novel case of trans-generational resistance in plants. PMID:24834319

  12. Vertical transmission of fungal endophytes is widespread in forbs.

    PubMed

    Hodgson, Susan; de Cates, Catherine; Hodgson, Joshua; Morley, Neil J; Sutton, Brian C; Gange, Alan C

    2014-04-01

    To date, it has been thought that endophytic fungi in forbs infect the leaves of their hosts most commonly by air-borne spores (termed "horizontal transmission"). Here, we show that vertical transmission from mother plant to offspring, via seeds, occurs in six forb species (Centaurea cyanus, C. nigra,Papaver rhoeas,Plantago lanceolata,Rumex acetosa, and Senecio vulgaris), suggesting that this may be a widespread phenomenon. Mature seeds were collected from field-grown plants and endophytes isolated from these, and from subsequent cotyledons and true leaves of seedlings, grown in sterile conditions. Most seeds contain one species of fungus, although the identity of the endophyte differs between plant species. Strong evidence for vertical transmission was found for two endophyte species, Alternaria alternata and Cladosporium sphaerospermum. These fungi were recovered from within seeds, cotyledons, and true leaves, although the plant species they were associated with differed. Vertical transmission appears to be an imperfect process, and germination seems to present a bottleneck for fungal growth. We also found that A. alternata and C. sphaerospermum occur on, and within pollen grains, showing that endophyte transmission can be both within and between plant generations. Fungal growth with the pollen tube is likely to be the way in which endophytes enter the developing seed. The fact that true vertical transmission seems common suggests a more mutualistic association between these fungi and their hosts than has previously been thought, and possession of endophytes by seedling plants could have far-reaching ecological consequences. Seedlings may have different growth rates and be better protected against herbivores and pathogens, dependent on the fungi that were present in the mother plant. This would represent a novel case of trans-generational resistance in plants. PMID:24834319

  13. Diverse bacteria inhabit living hyphae of phylogenetically diverse fungal endophytes.

    PubMed

    Hoffman, Michele T; Arnold, A Elizabeth

    2010-06-01

    Both the establishment and outcomes of plant-fungus symbioses can be influenced by abiotic factors, the interplay of fungal and plant genotypes, and additional microbes associated with fungal mycelia. Recently bacterial endosymbionts were documented in soilborne Glomeromycota and Mucoromycotina and in at least one species each of mycorrhizal Basidiomycota and Ascomycota. Here we show for the first time that phylogenetically diverse endohyphal bacteria occur in living hyphae of diverse foliar endophytes, including representatives of four classes of Ascomycota. We examined 414 isolates of endophytic fungi, isolated from photosynthetic tissues of six species of cupressaceous trees in five biogeographic provinces, for endohyphal bacteria using microscopy and molecular techniques. Viable bacteria were observed within living hyphae of endophytic Pezizomycetes, Dothideomycetes, Eurotiomycetes, and Sordariomycetes from all tree species and biotic regions surveyed. A focus on 29 fungus/bacterium associations revealed that bacterial and fungal phylogenies were incongruent with each other and with taxonomic relationships of host plants. Overall, eight families and 15 distinct genotypes of endohyphal bacteria were recovered; most were members of the Proteobacteria, but a small number of Bacillaceae also were found, including one that appears to occur as an endophyte of plants. Frequent loss of bacteria following subculturing suggests a facultative association. Our study recovered distinct lineages of endohyphal bacteria relative to previous studies, is the first to document their occurrence in foliar endophytes representing four of the most species-rich classes of fungi, and highlights for the first time their diversity and phylogenetic relationships with regard both to the endophytes they inhabit and the plants in which these endophyte-bacterium symbiota occur. PMID:20435775

  14. Characterization of cellulases of fungal endophytes isolated from Espeletia spp.

    PubMed

    Cabezas, Luisa; Calderon, Carolina; Medina, Luis Miguel; Bahamon, Isabela; Cardenas, Martha; Bernal, Adriana Jimena; Gonzalez, Andrés; Restrepo, Silvia

    2012-12-01

    Endophytes are microorganisms that asymptomatically invade plant tissues. They can stimulate plant growth and/or provide defense against pathogen attacks through the production of secondary metabolites. Most endophyte species are still unknown, and because they may have several applications, the study of their metabolic capabilities is essential. We characterized 100 endophytes isolated from Espeletia spp., a genus unique to the paramo ecosystem, an extreme environment in the Andean mountain range. We evaluated the cellulolytic potential of these endophytes on the saccharification of the oil palm empty fruit bunch (OPEFB). The total cellulolytic activity was measured for each endophyte on filter paper (FPA). In addition, the specific carboxymethyl cellulase (CMCase), exoglucanase, and β-glucosidase activities were determined. We found four fungi positive for cellulases. Of these fungi, Penicillium glabrum had the highest cellulolytic activity after partial purification, with maximal CMCase, exoglucanase and β-glucosidase enzyme activities of 44.5, 48.3, and 0.45 U/ml, respectively. Our data showed that the bioprospection of fungi and the characterization of their enzymes may facilitate the process of biofuel production. PMID:23274988

  15. Quorum quenching is an antivirulence strategy employed by endophytic bacteria.

    PubMed

    Kusari, Parijat; Kusari, Souvik; Lamshöft, Marc; Sezgin, Selahaddin; Spiteller, Michael; Kayser, Oliver

    2014-08-01

    Bacteria predominantly use quorum sensing to regulate a plethora of physiological activities such as cell-cell crosstalk, mutualism, virulence, competence, biofilm formation, and antibiotic resistance. In this study, we investigated how certain potent endophytic bacteria harbored in Cannabis sativa L. plants use quorum quenching as an antivirulence strategy to disrupt the cell-to-cell quorum sensing signals in the biosensor strain, Chromobacterium violaceum. We used a combination of high-performance liquid chromatography high-resolution mass spectrometry (HPLC-ESI-HRMS(n)) and matrix-assisted laser desorption ionization imaging high-resolution mass spectrometry (MALDI-imaging-HRMS) to first quantify and visualize the spatial distribution of the quorum sensing molecules in the biosensor strain, C. violaceum. We then showed, both quantitatively and visually in high spatial resolution, how selected endophytic bacteria of C. sativa can selectively and differentially quench the quorum sensing molecules of C. violaceum. This study provides fundamental insights into the antivirulence strategies used by endophytes in order to survive in their ecological niches. Such defense mechanisms are evolved in order to thwart the plethora of pathogens invading associated host plants in a manner that prevents the pathogens from developing resistance against the plant/endophyte bioactive secondary metabolites. This work also provides evidence towards utilizing endophytes as tools for biological control of bacterial phytopathogens. In continuation, such insights would even afford new concepts and strategies in the future for combating drug resistant bacteria by quorum-inhibiting clinical therapies. PMID:24846733

  16. Endophytic Fungi Isolated from Coleus amboinicus Lour Exhibited Antimicrobial Activity

    PubMed Central

    Astuti, Puji; Sudarsono, Sudarsono; Nisak, Khoirun; Nugroho, Giri Wisnu

    2014-01-01

    Purpose: Coleus amboinicus is a medicinal plant traditionally used to treat various diseases such as throat infection, cough and fever, diarrhea, nasal congestion and digestive problems. The plant was explored for endophytic fungi producing antimicrobial agents. Methods: Screening for endophytic fungi producing antimicrobial agents was conducted using agar plug method and antimicrobial activity of promising ethyl acetate extracts was determined by disc diffusion assay. Thin layer chromatography (TLC) - bioautography was performed to localize the bioactive components within the extract. TLC visualization detection reagents were used to preliminary analyze phytochemical groups of the bioactive compounds. Results: Three endophytic fungi were obtained, two of them showed promising potential. Agar diffusion method showed that endophytic fungi CAL-2 exhibited antimicrobial activity against P. aeruginosa, B. subtilis, S. aureus and S. thypi, whilst CAS-1 inhibited the growth of B. subtilis. TLC bioautography of ethyl acetate extract of CAL-2 revealed at least three bands exhibited antimicrobial activity and at least two bands showed inhibition of B. subtilis growth. Preliminary analysis of the crude extracts suggests that bioactive compounds within CAL-2 extract are terpenoids, phenolics and phenyl propanoid compounds whilst the antimicrobial agents within CAS-1 extract are terpenoids, propylpropanoids, alkaloids or heterocyclic nitrogen compounds. Conclusion: These data suggest the potential of endophytic fungi of C. amboinicus as source for antimicrobial agents. PMID:25671195

  17. Endohyphal Bacterium Enhances Production of Indole-3-Acetic Acid by a Foliar Fungal Endophyte

    PubMed Central

    Hoffman, Michele T.; Gunatilaka, Malkanthi K.; Wijeratne, Kithsiri; Gunatilaka, Leslie; Arnold, A. Elizabeth

    2013-01-01

    Numerous plant pathogens, rhizosphere symbionts, and endophytic bacteria and yeasts produce the important phytohormone indole-3-acetic acid (IAA), often with profound effects on host plants. However, to date IAA production has not been documented among foliar endophytes -- the diverse guild of primarily filamentous Ascomycota that live within healthy, above-ground tissues of all plant species studied thus far. Recently bacteria that live within hyphae of endophytes (endohyphal bacteria) have been detected, but their effects have not been studied previously. Here we show not only that IAA is produced in vitro by a foliar endophyte (here identified as Pestalotiopsis aff. neglecta, Xylariales), but that IAA production is enhanced significantly when the endophyte hosts an endohyphal bacterium (here identified as Luteibacter sp., Xanthomonadales). Both the endophyte and the endophyte/bacterium complex appear to rely on an L-tryptophan dependent pathway for IAA synthesis. The bacterium can be isolated from the fungus when the symbiotic complex is cultivated at 36°C. In pure culture the bacterium does not produce IAA. Culture filtrate from the endophyte-bacterium complex significantly enhances growth of tomato in vitro relative to controls and to filtrate from the endophyte alone. Together these results speak to a facultative symbiosis between an endophyte and endohyphal bacterium that strongly influences IAA production, providing a new framework in which to explore endophyte-plant interactions. PMID:24086270

  18. Fungal endophytes as prolific source of phytochemicals and other bioactive natural products: A review.

    PubMed

    Nisa, Humeera; Kamili, Azra N; Nawchoo, Irshad A; Shafi, Sana; Shameem, Nowsheen; Bandh, Suhaib A

    2015-05-01

    Endophytic fungi are those that live internally in apparently healthy and asymptomatic hosts. Endophytic fungi appear to be ubiquitous; indeed, no study has yet shown the existence of a plant species without endophytes. High species diversity is another characteristic of endophytic mycobiota which is depicted by the fact that it is quite common for endophyte surveys to find assemblages consisting of more than 30 fungal species per host plant species. Medicinal plants had been used to isolate and characterize directly the bioactive metabolites. However, the discovery of fungal endophytes inside these plants with capacity to produce the same compounds shifted the focus of new drug sources from plants to fungi. Bioactive natural products from endophytic fungi, isolated from different plant species, are attracting considerable attention from natural product chemists and biologists alike which is clearly depicted by the steady increase of publications devoted to this topic during the recent years. This review will highlight the chemical potential of endophytic fungi with focus on the detection of pharmaceutically valuable plant constituents as products of fungal biosynthesis. In addition, it will cover newly discovered endophytic fungi and also new bioactive metabolites reported in recent years from fungal endophytes. It summarizes the up-to-date and comprehensive information on bioactive compounds from endophytic fungi by having done a thorough survey of literature. PMID:25865953

  19. Endophytic fungus improves growth and metal uptake of Lolium arundinaceum Darbyshire ex. Schreb.

    PubMed

    Ren, Anzhi; Li, Chuan; Gao, Yubao

    2011-03-01

    The effect of endophyte infection on plant growth, cadmium (Cd) uptake, and Cd translocation was investigated using tall fescue (Lolium arundinaceum) grown in greenhouses in contaminated solution. Endophyte infection significantly increased tiller number and biomass of the host grass under both control and Cd-stress conditions. Endophyte infection not only enhanced Cd accumulation in tall fescue, but also improved Cd transport from the root to the shoot. Under 20 mg L(-1) Cd stress, the phytoextraction efficiency of endophyte-infected (EI) tall fescue was 2.41-fold higher than endophyte-free plants. Although the total Cd accumulation in EI tall fescue was insufficient for practical phytoextraction applications, the observed high biomass production and tolerance of stress from abiotic factors including heavy metals, gives endophyte/plant associations the potential to be a model for endophyte-assisted phytoremediation of metal-polluted soils. PMID:21598789

  20. Bioprospecting for Microbial Endophytes and Their Natural Products

    PubMed Central

    Strobel, Gary; Daisy, Bryn

    2003-01-01

    Endophytic microorganisms are to be found in virtually every plant on earth. These organisms reside in the living tissues of the host plant and do so in a variety of relationships, ranging from symbiotic to slightly pathogenic. Because of what appears to be their contribution to the host plant, the endophytes may produce a plethora of substances of potential use to modern medicine, agriculture, and industry. Novel antibiotics, antimycotics, immunosuppressants, and anticancer compounds are only a few examples of what has been found after the isolation, culture, purification, and characterization of some choice endophytes in the recent past. The potential prospects of finding new drugs that may be effective candidates for treating newly developing diseases in humans, plants, and animals are great. PMID:14665674

  1. Fungal endophytes characterization from four species of Diplazium Swartz

    NASA Astrophysics Data System (ADS)

    Affina-Eliya, A. A.; Noraini, T.; Nazlina, I.; Ruzi, A. R.

    2014-09-01

    Four species on genus Diplazium namely Diplazium tomentosum, D. sorzogonense, D. asperum and D. accedens of Peninsular Malaysia were studied for presence of fungal endophyte. The objective of this study is to characterize fungal endophytes in the rhizome of four Diplazium species. The rhizome was surface sterilized and incubated to isolate fungal endophytes. Characterization of the colonies was performed by macroscopic morphological, microscopic identification, types of hyphae and mycelium, and spore structure. For isolation that produces spores, the structure of conidiophores and conidia were identified. From this study, four fungal have been isolated and determined as Aspergillus sp. (isolates AE 1), Aspergillus fumigatus (isolates AE 2), Aspergillus versicolor (isolates AE 3) and Verticillium sp. (isolates AE 4). The fungal isolates from this study were classified from the same family Moniliaceae.

  2. Construction of a shuttle vector for protein secretory expression in Bacillus subtilis and the application of the mannanase functional heterologous expression.

    PubMed

    Guo, Su; Tang, Jia-jie; Wei, Dong-zhi; Wei, Wei

    2014-04-01

    We report the construction of two Bacillus subtilis expression vectors, pBNS1/pBNS2. Both vectors are based on the strong promoter P43 and the ampicillin resistance gene expression cassette. Additionally, a fragment with the Shine-Dalgarno sequence and a multiple cloning site (BamHI, SalI, SacI, XhoI, PstI, SphI) were inserted. The coding region for the amyQ (encoding an amylase) signal peptide was fused to the promoter P43 of pBNS1 to construct the secreted expression vector pBNS2. The applicability of vectors was tested by first generating the expression vectors pBNS1-GFP/pBNS2-GFP and then detecting for green fluorescent protein gene expression. Next, the mannanase gene from B. pumilus Nsic-2 was fused to vector pBNS2 and we measured the mannanase activity in the supernatant. The mannanase total enzyme activity was 8.65 U/ml, which was 6 times higher than that of the parent strain. Our work provides a feasible way to achieve an effective transformation system for gene expression in B. subtilis and is the first report to achieve B. pumilus mannanase secretory expression in B. subtilis. PMID:24375416

  3. In planta reduction of maize seedling stalk lesions by the bacterial endophyte Bacillus mojavensis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Due to autoinfection and alloinfection, maize is susceptible to infection by Fusarium verticillioides resulting in diseases and contamination of maize kernels with the fumonisin mycotoxins. Attempts at controlling this fungus are currently being done with biocontrol agents such as bacteria and this...

  4. Ruling out Bacillus anthracis.

    PubMed

    Papaparaskevas, Joseph; Houhoula, Dimitra P; Papadimitriou, Maria; Saroglou, Georgios; Legakis, Nicholas J; Zerva, Loukia

    2004-04-01

    Optimization of methods for ruling out Bacillus anthracis leads to increased yields, faster turnaround times, and a lighter workload. We used 72 environmental non-B. anthracis bacilli to validate methods for ruling out B. anthracis. Most effective were the use of horse blood agar, motility testing after isolates had a 2-h incubation in trypticase soy broth, and screening isolates with a B. anthracis-selective agar. PMID:15200872

  5. Isolation, purification and characterization of a surfactants-, laundry detergents- and organic solvents-resistant alkaline protease from Bacillus sp. HR-08.

    PubMed

    Moradian, Fatemeh; Khajeh, Khosro; Naderi-Manesh, Hossein; Sadeghizadeh, Majid

    2009-10-01

    Bacillus sp. HR-08 screened from soil samples of Iran, is capable of producing proteolytic enzymes. 16S rDNA analysis showed that this strain is closely related to Bacillus subtilis, Bacillus licheniformis, Bacillus pumilus, Bacillus mojavensis, and Bacillus atrophaeus. The zymogram analysis of the crude extract revealed the presence of five extracellular proteases. One of the proteases was purified in three steps procedure involving ammonium sulfate precipitation, DEAE-Sepharose ionic exchange and Sephacryl S-200 gel filtration chromatography. The molecular mass of the enzyme on SDS-PAGE was estimated to be 29 kDa. The protease exhibited maximum activity at pH 10.0 and 60 degrees C and was inhibited by PMSF but it was not affected by cysteine inhibitors, suggesting that the enzyme is a serine alkaline protease. Irreversible thermoinactivation of enzyme was examined at 50, 60, and 70 degrees C in the presence of 10 mM CaCl(2). Results showed that the protease activity retains more than 80% and 50% of its initial activity after incubation for 30 min at 60 and 70 degrees C, respectively. This enzyme had good stability in the presence of H(2)O(2), nonionic surfactant, and local detergents and its activity was enhanced in the presence of 20% of dimethyl sulfoxide (DMSO), dimethyl formamide (DMF) and isopropanol. The enzyme retained more than 90% of its initial activity after pre-incubation 1 h at room temperature in the presence of 20% of these solvents. Also, activation can be seen for the enzyme at high concentration (50%, v/v) of DMF and DMSO. PMID:19148779

  6. Functional characterization of salicylate hydroxylase from the fungal endophyte Epichloë festucae.

    PubMed

    Ambrose, Karen V; Tian, Zipeng; Wang, Yifei; Smith, Jordan; Zylstra, Gerben; Huang, Bingru; Belanger, Faith C

    2015-01-01

    Epichloë spp. are symbiotic fungal endophytes of many cool season grasses. The presence of the fungal endophytes often confers insect, drought, and disease tolerance to the host grasses. The presence of the fungal endophytes within the host plants does not elicit host defense responses. The molecular basis for this phenomenon is not known. Epichloë festucae, the endophyte of Festuca rubra, expresses a salicylate hydroxylase similar to NahG from the bacterium Pseudomonas putida. Few fungal salicylate hydroxylase enzymes have been reported. The in planta expression of an endophyte salicylate hydroxylase raised the possibility that degradation of plant-produced salicylic acid is a factor in the mechanism of how the endophyte avoids eliciting host plant defenses. Here we report the characterization of the E. festucae salicylate hydroxylase, designated Efe-shyA. Although the fungal enzyme has the expected activity, based on salicylic acid levels in endophyte-free and endophyte-infected plants it is unlikely that expression of the endophyte salicylate hydroxylase is a factor in the lack of a host defense response to the presence of the fungal endophyte. PMID:26055188

  7. Functional characterization of salicylate hydroxylase from the fungal endophyte Epichloë festucae

    PubMed Central

    Ambrose, Karen V.; Tian, Zipeng; Wang, Yifei; Smith, Jordan; Zylstra, Gerben; Huang, Bingru; Belanger, Faith C.

    2015-01-01

    Epichloë spp. are symbiotic fungal endophytes of many cool season grasses. The presence of the fungal endophytes often confers insect, drought, and disease tolerance to the host grasses. The presence of the fungal endophytes within the host plants does not elicit host defense responses. The molecular basis for this phenomenon is not known. Epichloë festucae, the endophyte of Festuca rubra, expresses a salicylate hydroxylase similar to NahG from the bacterium Pseudomonas putida. Few fungal salicylate hydroxylase enzymes have been reported. The in planta expression of an endophyte salicylate hydroxylase raised the possibility that degradation of plant-produced salicylic acid is a factor in the mechanism of how the endophyte avoids eliciting host plant defenses. Here we report the characterization of the E. festucae salicylate hydroxylase, designated Efe-shyA. Although the fungal enzyme has the expected activity, based on salicylic acid levels in endophyte-free and endophyte-infected plants it is unlikely that expression of the endophyte salicylate hydroxylase is a factor in the lack of a host defense response to the presence of the fungal endophyte. PMID:26055188

  8. [Diversity and community structure of endophytic fungi from Taxus chinensis var. mairei].

    PubMed

    2014-07-01

    A total of 628 endophytic fungi were isolated from 480 tissue segments of needles and branches of Taxus chinensis var. mairei. According to morphological characteristics and ITS sequences, they represented 43 taxa in 28 genera, of which 10 Hyphomycetes, 20 Coelomycetes, 12 Ascomycetes and 1 unknown fungus. Phomopsis mali was confirmed as the dominant species. In accordance with relative frequency, Alternaria alternata, Aureobasidium pullulans, Colletotrichum boninense, C. gloeosporioides, Epicoccum nigrum , Fungal sp., Fusarium lateritium, Glomerella cingulata, Magnaporthales sp. , Nigrospora oryzae, Pestalotiopsis maculiformans, P. microspora, Peyronellaea glomerata and Xylaria sp. 1 were more common in T. chinensis var. mairei. T. chinensis var. mairei were severely infected by endophytic fungi. Endophytic fungi were found in 81 percent of plant tissues with a high diversity. Distribution ranges of endophytic fungi were influenced by tissue properties. The colonization rate, richness, diversity of endophytic fungi in needles were obviously lower than in branches, and kinds of endophytic fungi between branches were more similar than those in needles, thus endophytic fungi had tissue preference. In addition, tissue age influenced the community structure of endophytic fungi. The elder branch tissues were, the higher colonization rate, richness, diversity of endophytic fungi were. Systematic studying the diversity and community structure of endophytic fungi in T. chinensis var. mairei and clarifying their distribution regularity in plant tissues would offer basic data and scientific basis for their development and utilization. Discussing the presence of fungal pathogens in healthy plant tissues would be of positive significance for source protection of T. chinensis var. mairei. PMID:25345060

  9. Pervasive Effects of Wildfire on Foliar Endophyte Communities in Montane Forest Trees.

    PubMed

    Huang, Yu-Ling; Devan, M M Nandi; U'Ren, Jana M; Furr, Susan H; Arnold, A Elizabeth

    2016-02-01

    Plants in all terrestrial ecosystems form symbioses with endophytic fungi that inhabit their healthy tissues. How these foliar endophytes respond to wildfires has not been studied previously, but is important given the increasing frequency and intensity of severe wildfires in many ecosystems, and because endophytes can influence plant growth and responses to stress. The goal of this study was to examine effects of severe wildfires on endophyte communities in forest trees, with a focus on traditionally fire-dominated, montane ecosystems in the southwestern USA. We evaluated the abundance, diversity, and composition of endophytes in foliage of Juniperus deppeana (Cupressaceae) and Quercus spp. (Fagaceae) collected contemporaneously from areas affected by recent wildfire and paired areas not affected by recent fire. Study sites spanned four mountain ranges in central and southern Arizona. Our results revealed significant effects of fires on endophyte communities, including decreases in isolation frequency, increases in diversity, and shifts in community structure and taxonomic composition among endophytes of trees affected by recent fires. Responses to fire were similar in endophytes of each host in these fire-dominated ecosystems and reflect regional fire-return intervals, with endophytes after fire representing subsets of the regional mycoflora. Together, these findings contribute to an emerging perspective on the responses of diverse communities to severe fire, and highlight the importance of considering fire history when estimating endophyte diversity and community structure for focal biomes. PMID:26370111

  10. Fungal Endophytes from Three Cultivars of Panax ginseng Meyer Cultivated in Korea.

    PubMed

    Park, Sang Un; Lim, Hyoun-Sub; Park, Kee-Choon; Park, Young-Hwan; Bae, Hanhong

    2012-01-01

    In order to investigate the diversity of endophytes, fungal endophytes in Panax ginseng Meyer cultivated in Korea were isolated and identified using internal transcribed spacer (ITS) sequences of ribosomal DNA. Three cultivars of 3-year-old ginseng roots (Chunpoong, Yunpoong, and Gumpoong) were used to isolate fungal endophytes. Surface sterilized ginseng roots were placed on potato dextrose agar plates supplemented with ampicilin and streptomycin to inhibit bacterial growth. Overall, 38 fungal endophytes were isolated from 12 ginseng roots. According to the sequence analysis of the ITS1-5.8S-ITS2, 38 fungal isolates were classified into 4 different fungal species, which were Phoma radicina, Fusarium oxysporum, Setophoma terrestris and Ascomycota sp. 2-RNK. The most dominant fungal endophyte was P. radicina in 3 cultivars. The percentage of dominant endophytes of P. radicina was 65.8%. The percentage of colonization frequency of P. radicina was 80%, 52.9%, and 75% in Chunpoong, Yunpoong, and Gumpoong, respectively. The second most dominant fungal endophyte was F. oxysporum. The diversity of the fungal endophytes was low and no ginseng cultivar specificity among endophytes was detected in this study. The identified endophytes can be potential fungi for the production of bioactive compounds and control against ginseng pathogens. PMID:23717111

  11. Endophytic bacteria: prospects and applications for the phytoremediation of organic pollutants.

    PubMed

    Afzal, Muhammad; Khan, Qaiser M; Sessitsch, Angela

    2014-12-01

    Recently, there has been an increased effort to enhance the efficacy of phytoremediation of contaminated environments by exploiting plant-microbe interactions. The combined use of plants and endophytic bacteria is an emerging approach for the clean-up of soil and water polluted with organic compounds. In plant-endophyte partnerships, plants provide the habitat as well as nutrients to their associated endophytic bacteria. In response, endophytic bacteria with appropriate degradation pathways and metabolic activities enhance degradation of organic pollutants, and diminish phytotoxicity and evapotranspiration of organic pollutants. Moreover, endophytic bacteria possessing plant growth-promoting activities enhance the plant's adaptation and growth in soil and water contaminated with organic pollutants. Overall, the application of endophytic bacteria gives new insights into novel protocols to improve phytoremediation efficiency. However, successful application of plant-endophyte partnerships for the clean-up of an environment contaminated with organic compounds depends on the abundance and activity of the degrading endophyte in different plant compartments. Although many endophytic bacteria have the potential to degrade organic pollutants and improve plant growth, their contribution to enhance phytoremediation efficiency is still underestimated. A better knowledge of plant-endophyte interactions could be utilized to increase the remediation of polluted soil environments and to protect the foodstuff by decreasing agrochemical residues in food crops. PMID:25078615

  12. Differential detection of a surrogate biological threat agent (Bacillus globigii) with a portable surface plasmon resonance biosensor.

    PubMed

    Adducci, Benjamin A; Gruszewski, Hope A; Khatibi, Piyum A; Schmale, David G

    2016-04-15

    New methods and technology are needed to quickly and accurately detect potential biological warfare agents, such as Bacillus anthracis, causal agent of anthrax in humans and animals. Here, we report the detection of a simulant of B. anthracis (B. globigii) alone and in a mixture with a different species of Bacillus to test non-specific interference using a portable surface plasmon resonance (SPR) biosensor (SPIRIT 4.0, Seattle Sensor Systems). Both direct capture and antibody amplification were used to determine the limit of detection for spores of B. globigii, and to detect spores of B. globigii in a mixed sample containing another Bacillus spp. Spores of B. globigii were detected by anti-B. globigii (anti-Bg) coated sensors by direct capture at a concentration of 10(7)spores/mL, and with a secondary antibody amplification at a concentration of 10(5)spores/mL. Spores of B. globigii were differentially detected in a 1:1 mixture with B. pumilus spores from equal concentrations (10(7)spores/mL) with a secondary antibody amplification. To our knowledge, this is the first report of the differential detection of B. globigii with SPR in a mixed sample containing at least one additional Bacillus spp., highlighting the potential for SPR to detect any target bacterium in a mixed sample of closely related species. With the availability of portable instrumentation to accurately detect biological warfare agents such as B. anthracis, emergency responders can implement protocols in a timely fashion, limiting the amount of exposed individuals. PMID:26606307

  13. Detection of Uncultured Seed Borne Endophytes in Atriplex canesens: Methods used in: “Seed Borne Endophyte Microbiomes are Associated with Vascular Plants”

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Micropropagated systems provide controlled habitats in which complex plant-endophyte interactions can be studied with minimal interference from superficial microbes. The series of protocols described herein can be used to examine seed borne endophyte consortia associated with woody plant species. ...

  14. Using Orchardgrass and Endophyte-Free Fescue Versus Endophyte-Infected Fescue Overseeded on Bermudagrass for Cow Herds: I. Four-Year Summary of Forage Characteristics

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A systems trial was designed to evaluate forage characteristics within mixed-species pastures consisting of: i) endophyte-infected tall fescue (Festuca arundinacea Schreb.; E+) mixed with common bermudagrass [Cynodon dactylon (L.) Pers.] and other forages; ii) endophyte-free tall fescue (E-) oversee...

  15. Defensive functions and responsible metabolites of microbial endophytes

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Increasing evidence indicates that plant microbiomes are influence by ecological successes of plant hosts. Further, endophytic microbes such as bacteria and fungi greatly affect plant stress tolerance and are responsible for defensive reaction to several forms of herbivory. What is not yet clear i...

  16. Bacterial endophytic communities in the grapevine depend on pest management.

    PubMed

    Campisano, Andrea; Antonielli, Livio; Pancher, Michael; Yousaf, Sohail; Pindo, Massimo; Pertot, Ilaria

    2014-01-01

    Microbial plant endophytes are receiving ever-increasing attention as a result of compelling evidence regarding functional interaction with the host plant. Microbial communities in plants were recently reported to be influenced by numerous environmental and anthropogenic factors, including soil and pest management. In this study we used automated ribosomal intergenic spacer analysis (ARISA) fingerprinting and pyrosequencing of 16S rDNA to assess the effect of organic production and integrated pest management (IPM) on bacterial endophytic communities in two widespread grapevines cultivars (Merlot and Chardonnay). High levels of the dominant Ralstonia, Burkholderia and Pseudomonas genera were detected in all the samples We found differences in the composition of endophytic communities in grapevines cultivated using organic production and IPM. Operational taxonomic units (OTUs) assigned to the Mesorhizobium, Caulobacter and Staphylococcus genera were relatively more abundant in plants from organic vineyards, while Ralstonia, Burkholderia and Stenotrophomonas were more abundant in grapevines from IPM vineyards. Minor differences in bacterial endophytic communities were also found in the grapevines of the two cultivars. PMID:25387008

  17. Influence of endophyte genotype on swainsonine concentrations in Oxytropis sericea

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Locoism is a toxic syndrome of livestock caused by the ingestion of a subset of legumes belonging to the Astragalus and Oxytropis genera known as “locoweeds”. Locoweeds contain the toxic indolizidine alkaloid swainsonine, which is produced by the endophytic fungi Undifilum species. Previously we r...

  18. Bioaugmentation with engineered endophytic bacteria improves contaminant fate in phytoremediation

    SciTech Connect

    Weyens, N.; van der Lelie, D.; Artois, T.; Smeets, K.; Taghavi, S.; Newman, L.; Carleer, R.; Vangronsveld, J.

    2009-12-01

    Phytoremediation of volatile organic contaminants often proves not ideal because plants and their rhizosphere microbes only partially degrade these compounds. Consequently, plants undergo evapotranspiration that contaminates the ambient air and, thus, undermines the merits of phytoremediation. Under laboratory conditions, endophytic bacteria equipped with the appropriate degradation pathways can improve in plant degradation of volatile organic contaminants. However, several obstacles must be overcome before engineered endophytes will be successful in field-scale phytoremediation projects. Here we report the first in situ inoculation of poplar trees, growing on a TCE-contaminated site, with the TCE-degrading strain Pseudomonas putida W619-TCE. In situ bioaugmentation with strain W619-TCE reduced TCE evapotranspiration by 90% under field conditions. This encouraging result was achieved after the establishment and enrichment of P. putida W619-TCE as a poplar root endophyte and by further horizontal gene transfer of TCE metabolic activity to members of the poplar's endogenous endophytic population. Since P. putida W619-TCE was engineered via horizontal gene transfer, its deliberate release is not restricted under European genetically modified organisms (GMO) regulations.

  19. Extracellular Hemicellulolytic Enzymes from the Maize Endophyte Acremonium zeae

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The maize endophyte Acremonium zeae was examined for production of extracellular enzymes that hydrolyze cellulose and hemicellulose. The most prominent enzyme activity in cell-free culture media from A. zeae NRRL 6415 was xylanase, with a specific activity of 60 U/mg from cultures grown on crude co...

  20. Elucidating biotic factors that influence assembly of fungal endophyte communities

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Most plants harbor a diverse assemblage of non-mycorrhizal fungal endophytes. These fungi can directly influence the host plant, and can instigate trophic cascades that affect surrounding communities of herbivores, plants, and animals. Despite this, biotic mechanisms that influence assembly of funga...

  1. TALL FESCUE AND ASSOCIATED MUTUALISTIC TOXIC FUNGAL ENDOPHYTES IN AGROECOSYSTEMS

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Bacon et al. (1977) proposed and Hoveland et al. (1980, 1986) substantiated that impaired health and performance of livestock grazing tall fescue [Lolium arundinaceum S. J. Darbyshire, formerly Festuca arundinacea Schreb.] were associated with the fungal endophyte Neotyphodium coenophialum Glenn, Ha...

  2. Potential roles for microbial endophytes in herbicide tolerance in plants.

    PubMed

    Tétard-Jones, Catherine; Edwards, Robert

    2016-02-01

    Herbicide tolerance in crops and weeds is considered to be monotrophic, i.e. determined by the relative susceptibility of the physiological process targeted and the plant's ability to metabolise and detoxify the agrochemical. A growing body of evidence now suggests that endophytes, microbes that inhabit plant tissues and provide a range of growth, health and defence enhancements, can contribute to other types of abiotic and biotic stress tolerance. The current evidence for herbicide tolerance being bitrophic, with both free-living and plant-associated endophytes contributing to tolerance in the host plant, has been reviewed. We propose that endophytes can directly contribute to herbicide detoxification through their ability to metabolise xenobiotics. In addition, we explore the paradigm that microbes can 'prime' resistance mechanisms in plants such that they enhance herbicide tolerance by inducing the host's stress responses to withstand the downstream toxicity caused by herbicides. This latter mechanism has the potential to contribute to the growth of non-target-site-based herbicide resistance in weeds. Microbial endophytes already contribute to herbicide detoxification in planta, and there is now significant scope to extend these interactions using synthetic biology approaches to engineer new chemical tolerance traits into crops via microbial engineering. © 2015 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry. PMID:26350619

  3. Endophytic bacteria in potato tubers affected by zebra chip disease

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Potato zebra chip disease (ZCD) could drastically reduce quality and value of all market classes of potato, costing growers and processors millions of dollars in losses in North America. Endophytic bacteria colonize the internal tissue and could have both positive and negative effects on host plants...

  4. Plant growth promotion induced by phosphate solubilizing endophytic Pseudomonas isolates.

    PubMed

    Oteino, Nicholas; Lally, Richard D; Kiwanuka, Samuel; Lloyd, Andrew; Ryan, David; Germaine, Kieran J; Dowling, David N

    2015-01-01

    The use of plant growth promoting bacterial inoculants as live microbial biofertilizers provides a promising alternative to chemical fertilizers and pesticides. Inorganic phosphate solubilization is one of the major mechanisms of plant growth promotion by plant associated bacteria. This involves bacteria releasing organic acids into the soil which solubilize the phosphate complexes converting them into ortho-phosphate which is available for plant up-take and utilization. The study presented here describes the ability of endophytic bacteria to produce gluconic acid (GA), solubilize insoluble phosphate, and stimulate the growth of Pisum sativum L. plants. This study also describes the genetic systems within three of these endophyte strains thought to be responsible for their effective phosphate solubilizing abilities. The results showed that many of the endophytic strains produced GA (14-169 mM) and have moderate to high phosphate solubilization capacities (~400-1300 mg L(-1)). When inoculated into P. sativum L. plants grown in soil under soluble phosphate limiting conditions, the endophytes that produced medium-high levels of GA displayed beneficial plant growth promotion effects. PMID:26257721

  5. Impact of endophytic microorganisms on plants, environment and humans.

    PubMed

    Nair, Dhanya N; Padmavathy, S

    2014-01-01

    Endophytes are microorganisms (bacteria or fungi or actinomycetes) that dwell within robust plant tissues by having a symbiotic association. They are ubiquitously associated with almost all plants studied till date. Some commonly found endophytes are those belonging to the genera Enterobacter sp., Colletotrichum sp., Phomopsis sp., Phyllosticta sp., Cladosporium sp., and so forth. Endophytic population is greatly affected by climatic conditions and location where the host plant grows. They produce a wide range of compounds useful for plants for their growth, protection to environmental conditions, and sustainability, in favour of a good dwelling place within the hosts. They protect plants from herbivory by producing certain compounds which will prevent animals from further grazing on the same plant and sometimes act as biocontrol agents. A large amount of bioactive compounds produced by them not only are useful for plants but also are of economical importance to humans. They serve as antibiotics, drugs or medicines, or the compounds of high relevance in research or as compounds useful to food industry. They are also found to have some important role in nutrient cycling, biodegradation, and bioremediation. In this review, we have tried to comprehend different roles of endophytes in plants and their significance and impacts on man and environment. PMID:24587715

  6. Biosynthetic Potential of Phylogenetically Unique Endophytic Actinomycetes from Tropical Plants? †

    PubMed Central

    Janso, Jeffrey E.; Carter, Guy T.

    2010-01-01

    The culturable diversity of endophytic actinomycetes associated with tropical, native plants is essentially unexplored. In this study, 123 endophytic actinomycetes were isolated from tropical plants collected from several locations in Papua New Guinea and Mborokua Island, Solomon Islands. Isolates were found to be prevalent in roots but uncommon in leaves. Initially, isolates were dereplicated to the strain level by ribotyping. Subsequent characterization of 105 unique strains by 16S rRNA gene sequence analysis revealed that 17 different genera were represented, and rare genera, such as Sphaerisporangium and Planotetraspora, which have never been previously reported to be endophytic, were quite prevalent. Phylogenetic analyses grouped many of the strains into clades distinct from known genera within Thermomonosporaceae and Micromonosporaceae, indicating that they may be unique genera. Bioactivity testing and liquid chromatography-mass spectrometry (LC-MS) profiling of crude fermentation extracts were performed on 91 strains. About 60% of the extracts exhibited bioactivity or displayed LC-MS profiles with spectra indicative of secondary metabolites. The biosynthetic potential of 29 nonproductive strains was further investigated by the detection of putative polyketide synthase (PKS) and nonribosomal peptide synthetase (NRPS) genes. Despite their lack of detectable secondary metabolite production in fermentation, most were positive for type I (66%) and type II (79%) PKS genes, and all were positive for NRPS genes. These results suggest that tropical plants from New Guinea and the adjacent archipelago are hosts to unique endophytic actinomycetes that possess significant biosynthetic potential. PMID:20472734

  7. Endophytic Phomopsis species: host range and implications for diversity estimates.

    PubMed

    Murali, T S; Suryanarayanan, T S; Geeta, R

    2006-07-01

    Foliar endophyte assemblages of teak trees growing in dry deciduous and moist deciduous forests of Nilgiri Biosphere Reserve were compared. A species of Phomopsis dominated the endophyte assemblages of teak, irrespective of the location of the host trees. Internal transcribed spacer sequence analysis of 11 different Phomopsis isolates (ten from teak and one from Cassia fistula) showed that they fall into two groups, which are separated by a relatively long branch that is strongly supported. The results showed that this fungus is not host restricted and that it continues to survive as a saprotroph in teak leaf, possibly by exploiting senescent leaves as well as the litter. Although the endophyte assemblage of a teak tree growing about 500 km from the forests was also dominated by a Phomopsis sp., it separated into a different group based on internal transcribed spacer sequence analysis. Our results with an endophytic Phomopsis sp. reinforce the earlier conclusions reached by others for pathogenic Phomopsis sp., i.e., that this fungus is not host specific, and the species concept of Phomopsis needs to be redefined. PMID:16917524

  8. Impact of Endophytic Microorganisms on Plants, Environment and Humans

    PubMed Central

    Nair, Dhanya N.; Padmavathy, S.

    2014-01-01

    Endophytes are microorganisms (bacteria or fungi or actinomycetes) that dwell within robust plant tissues by having a symbiotic association. They are ubiquitously associated with almost all plants studied till date. Some commonly found endophytes are those belonging to the genera Enterobacter sp., Colletotrichum sp., Phomopsis sp., Phyllosticta sp., Cladosporium sp., and so forth. Endophytic population is greatly affected by climatic conditions and location where the host plant grows. They produce a wide range of compounds useful for plants for their growth, protection to environmental conditions, and sustainability, in favour of a good dwelling place within the hosts. They protect plants from herbivory by producing certain compounds which will prevent animals from further grazing on the same plant and sometimes act as biocontrol agents. A large amount of bioactive compounds produced by them not only are useful for plants but also are of economical importance to humans. They serve as antibiotics, drugs or medicines, or the compounds of high relevance in research or as compounds useful to food industry. They are also found to have some important role in nutrient cycling, biodegradation, and bioremediation. In this review, we have tried to comprehend different roles of endophytes in plants and their significance and impacts on man and environment. PMID:24587715

  9. Bacterial Endophytic Communities in the Grapevine Depend on Pest Management

    PubMed Central

    Campisano, Andrea; Antonielli, Livio; Pancher, Michael; Yousaf, Sohail; Pindo, Massimo; Pertot, Ilaria

    2014-01-01

    Microbial plant endophytes are receiving ever-increasing attention as a result of compelling evidence regarding functional interaction with the host plant. Microbial communities in plants were recently reported to be influenced by numerous environmental and anthropogenic factors, including soil and pest management. In this study we used automated ribosomal intergenic spacer analysis (ARISA) fingerprinting and pyrosequencing of 16S rDNA to assess the effect of organic production and integrated pest management (IPM) on bacterial endophytic communities in two widespread grapevines cultivars (Merlot and Chardonnay). High levels of the dominant Ralstonia, Burkholderia and Pseudomonas genera were detected in all the samples We found differences in the composition of endophytic communities in grapevines cultivated using organic production and IPM. Operational taxonomic units (OTUs) assigned to the Mesorhizobium, Caulobacter and Staphylococcus genera were relatively more abundant in plants from organic vineyards, while Ralstonia, Burkholderia and Stenotrophomonas were more abundant in grapevines from IPM vineyards. Minor differences in bacterial endophytic communities were also found in the grapevines of the two cultivars. PMID:25387008

  10. Plant growth promotion induced by phosphate solubilizing endophytic Pseudomonas isolates

    PubMed Central

    Oteino, Nicholas; Lally, Richard D.; Kiwanuka, Samuel; Lloyd, Andrew; Ryan, David; Germaine, Kieran J.; Dowling, David N.

    2015-01-01

    The use of plant growth promoting bacterial inoculants as live microbial biofertilizers provides a promising alternative to chemical fertilizers and pesticides. Inorganic phosphate solubilization is one of the major mechanisms of plant growth promotion by plant associated bacteria. This involves bacteria releasing organic acids into the soil which solubilize the phosphate complexes converting them into ortho-phosphate which is available for plant up-take and utilization. The study presented here describes the ability of endophytic bacteria to produce gluconic acid (GA), solubilize insoluble phosphate, and stimulate the growth of Pisum sativum L. plants. This study also describes the genetic systems within three of these endophyte strains thought to be responsible for their effective phosphate solubilizing abilities. The results showed that many of the endophytic strains produced GA (14–169 mM) and have moderate to high phosphate solubilization capacities (~400–1300 mg L−1). When inoculated into P. sativum L. plants grown in soil under soluble phosphate limiting conditions, the endophytes that produced medium-high levels of GA displayed beneficial plant growth promotion effects. PMID:26257721

  11. An endophytic Coniochaeta velutina producing broad spectrum antimycotics.

    PubMed

    Xie, Jie; Strobel, Gary A; Feng, Tao; Ren, Huishuang; Mends, Morgan T; Zhou, Zeyang; Geary, Brad

    2015-06-01

    An endophyte (PC27-5) was isolated from stem tissue of Western hemlock (Tsuga heterophylla) in a Pacific Northwest temperate rainforest. Phylogenetic analyses, based on ITS-5.8S rDNA and 18S rDNA sequence data, combined with cultural and morphological analysis showed that endophyte PC27-5 exhibited all characteristics of a fungus identical to Coniochaeta velutina. Furthermore, wide spectrum antimycotics were produced by this endophyte that were active against such plant pathogens as Sclerotinia sclerotiorum, Pythium ultimum, and Verticillium dahliae and lethal to Phythophthora cinnamomi, Pythium ultimum, and Phytophthora palmivora in plate tests. The bioactive components were purified through organic solvent extraction, followed by silica column chromatography, and finally preparative HPLC. The minimum inhibitory concentration of the active fraction to Pythium ultimum, which was gained from preparative HPLC, was 11 ?g/ml. UPLC-HRMS analysis showed there were two similar components in the antimycotic fraction. Their molecular formulae were established as C30H22O11 (compound I) and C30H22O10 (compound II) respectively, and preliminary spectral results indicate that they are anthroquinone glycosides. Other non-biologically active compounds were identified in culture fluids of this fungus by spectral means as emodin and chrysophanol--anthroquinone derivatives. This is the first report that Coniochaeta velutina as an endophyte produces bioactive antifungal components. PMID:26025171

  12. Effects of natural hybrid and non-hybrid Epichloë endophytes on the response of Hordelymus europaeus to drought stress.

    PubMed

    Oberhofer, Martina; Güsewell, Sabine; Leuchtmann, Adrian

    2014-01-01

    Interspecific hybrid endophytes of the genus Epichloë (Ascomycota, Clavicipitaceae) are prevalent in wild grass populations, possibly because of their larger gene variation, resulting in increased fitness benefits for host plants; however, the reasons are not yet known. We tested hypotheses regarding niche expansion mediated by hybrid endophytes, population-dependent interactions and local co-adaptation in the woodland grass Hordelymus europaeus, which naturally hosts both hybrid and non-hybrid endophyte taxa. Seedlings derived from seeds of four grass populations made endophyte free were re-inoculated with hybrid or non-hybrid endophyte strains, or left endophyte free. Plants were grown in the glasshouse with or without drought treatment. Endophyte infection increased plant biomass and tiller production by 10-15% in both treatments. Endophyte types had similar effects on growth, but opposite effects on reproduction: non-hybrid endophytes increased seed production, whereas hybrid endophytes reduced or prevented it completely. The results are consistent with the observation that non-hybrid endophytes in H. europaeus prevail at dry sites, but cannot explain the prevalence of hybrid endophytes. Thus, our results do not support the hypothesis of niche expansion of hybrid-infected plants. Moreover, plants inoculated with native relative to foreign endophytes yielded higher infections, but both showed similar growth and survival, suggesting weak co-adaptation. PMID:24102453

  13. Leaf endophyte load influences fungal garden development in leaf-cutting ants

    PubMed Central

    2012-01-01

    Background Previous work has shown that leaf-cutting ants prefer to cut leaf material with relatively low fungal endophyte content. This preference suggests that fungal endophytes exact a cost on the ants or on the development of their colonies. We hypothesized that endophytes may play a role in their host plants’ defense against leaf-cutting ants. To measure the long-term cost to the ant colony of fungal endophytes in their forage material, we conducted a 20-week laboratory experiment to measure fungal garden development for colonies that foraged on leaves with low or high endophyte content. Results Colony mass and the fungal garden dry mass did not differ significantly between the low and high endophyte feeding treatments. There was, however, a marginally significant trend toward greater mass of fungal garden per ant worker in the low relative to the high endophyte treatment. This trend was driven by differences in the fungal garden mass per worker from the earliest samples, when leaf-cutting ants had been foraging on low or high endophyte leaf material for only 2 weeks. At two weeks of foraging, the mean fungal garden mass per worker was 77% greater for colonies foraging on leaves with low relative to high endophyte loads. Conclusions Our data suggest that the cost of endophyte presence in ant forage material may be greatest to fungal colony development in its earliest stages, when there are few workers available to forage and to clean leaf material. This coincides with a period of high mortality for incipient colonies in the field. We discuss how the endophyte-leaf-cutter ant interaction may parallel constitutive defenses in plants, whereby endophytes reduce the rate of colony development when its risk of mortality is greatest. PMID:23140096

  14. Effects of the Epichloë fungal endophyte symbiosis with Schedonorus pratensis on host grass invasiveness.

    PubMed

    Shukla, Kruti; Hager, Heather A; Yurkonis, Kathryn A; Newman, Jonathan A

    2015-07-01

    Initial studies of grass-endophyte mutualisms using Schedonorus arundinaceus cultivar Kentucky-31 infected with the vertically transmitted endophyte Epichloë coenophiala found strong, positive endophyte effects on host-grass invasion success. However, more recent work using different cultivars of S. arundinaceus has cast doubt on the ubiquity of this effect, at least as it pertains to S. arundinaceus-E. coenophiala. We investigated the generality of previous work on vertically transmitted Epichloë-associated grass invasiveness by studying a pair of very closely related species: S. pratensis and E. uncinata. Seven cultivars of S. pratensis and two cultivars of S. arundinaceus that were developed with high- or low-endophyte infection rate were broadcast seeded into 2 × 2-m plots in a tilled, old-field grassland community in a completely randomized block design. Schedonorus abundance, endophyte infection rate, and co-occurring vegetation were sampled 3, 4, 5, and 6 years after establishment, and the aboveground invertebrate community was sampled in S. pratensis plots 3 and 4 years after establishment. Endophyte infection did not enable the host grass to achieve high abundance in the plant community. Contrary to expectations, high-endophyte S. pratensis increased plant richness relative to low-endophyte cultivars. However, as expected, high-endophyte S. pratensis marginally decreased invertebrate taxon richness. Endophyte effects on vegetation and invertebrate community composition were inconsistent among cultivars and were weaker than temporal effects. The effect of the grass-Epichloë symbiosis on diversity is not generalizable, but rather specific to species, cultivar, infection, and potentially site. Examining grass-endophyte systems using multiple cultivars and species replicated among sites will be important to determine the range of conditions in which endophyte associations benefit host grass performance and have subsequent effects on co-occurring biotic communities. PMID:26257873

  15. Effects of the Epichloë fungal endophyte symbiosis with Schedonorus pratensis on host grass invasiveness

    PubMed Central

    Shukla, Kruti; Hager, Heather A; Yurkonis, Kathryn A; Newman, Jonathan A

    2015-01-01

    Initial studies of grass–endophyte mutualisms using Schedonorus arundinaceus cultivar Kentucky-31 infected with the vertically transmitted endophyte Epichloë coenophiala found strong, positive endophyte effects on host-grass invasion success. However, more recent work using different cultivars of S. arundinaceus has cast doubt on the ubiquity of this effect, at least as it pertains to S. arundinaceus–E. coenophiala. We investigated the generality of previous work on vertically transmitted Epichloë-associated grass invasiveness by studying a pair of very closely related species: S. pratensis and E. uncinata. Seven cultivars of S. pratensis and two cultivars of S. arundinaceus that were developed with high- or low-endophyte infection rate were broadcast seeded into 2 × 2-m plots in a tilled, old-field grassland community in a completely randomized block design. Schedonorus abundance, endophyte infection rate, and co-occurring vegetation were sampled 3, 4, 5, and 6 years after establishment, and the aboveground invertebrate community was sampled in S. pratensis plots 3 and 4 years after establishment. Endophyte infection did not enable the host grass to achieve high abundance in the plant community. Contrary to expectations, high-endophyte S. pratensis increased plant richness relative to low-endophyte cultivars. However, as expected, high-endophyte S. pratensis marginally decreased invertebrate taxon richness. Endophyte effects on vegetation and invertebrate community composition were inconsistent among cultivars and were weaker than temporal effects. The effect of the grass–Epichloë symbiosis on diversity is not generalizable, but rather specific to species, cultivar, infection, and potentially site. Examining grass–endophyte systems using multiple cultivars and species replicated among sites will be important to determine the range of conditions in which endophyte associations benefit host grass performance and have subsequent effects on co-occurring biotic communities. PMID:26257873

  16. Identification and characterization of the Sudanese Bacillus thuringiensis and related bacterial strains for their efficacy against Helicoverpa armigera and Tribolium castaneum.

    PubMed

    Gorashi, N E; Tripathi, M; Kalia, V; Gujar, G T

    2014-06-01

    Forty-four isolates of Bacillus thuringiensis like bacteria from various sources in different locations from Sudan were tested for their insecticidal activity. The toxicity of these isolates ranged from 6.6 to 70% to the neonates of cotton bollworm, Helicoverpa armigera at 10 ppm concentration. The most effective ones are Kb-29, St-6 and Wh-1 comparable with HD-1. Toxicity of isolates to larvae of the red flour beetle, Tribolium castaneum ranged from 20 to 100%. Isolates St-2 and St-23 gave 100% larval mortality within 15 days of exposure and were at par with Ab-8, Ab-12, Kb-26, Kb-30, Om-4, Po-2, Po-5, Po-7, Sa-8 and Wh-5 and were also comparable with E. coli clone expressing Cry3 toxin. The most effective five isolates viz., Kb-29, St-2, St-6, St-23 and Wh-1 belonged to B. thuringiensis. The St-6 isolate, which also showed high toxicity to T. castaneum larvae, had cry1 genes along with coleopteran active cry28 genes, but not cry3 genes. Of the 25 isolates characterized with 16s DNA sequencing, seven belonged to Paenibacillus spp., one Lysinibacillus sphaericus, one Bacillus pumilus, four Bacillus spp., and rest 12 belonged to B. thuringiensis. Biochemical characterization in each species showed variation. The present study shows potential of some isolates like Kb-29, St-2, St-6, St-23 and Wh-1 as promising bioinsecticides. PMID:24956895

  17. Development of a versatile cassette for directional genome walking using cassette ligation-mediated PCR and its application in the cloning of complete lipolytic genes from Bacillus species.

    PubMed

    Nthangeni, Mulalo B; Ramagoma, Faranani; Tlou, Matsobane G; Litthauer, Derek

    2005-05-01

    Since the invention of the PCR technology, adaptation techniques to clone DNA fragments flanking the known sequence continue to be developed. We describe a perfectly annealed cassette available in almost unlimited quantities with variable sticky-and blunt-end restriction enzyme recognition sites for efficient restriction and ligation with the restricted target genomic DNA. The cassette provides a 200-bp sequence, which is used to design a variety of cassette-specific primers. The dephosphorylation prevents cassette self-ligation and creates a nick at the cassette: target genome DNA ligation site suppressing unspecific PCR amplifications. We introduce the single-strand amplification PCR (SSA-PCR) technique where a lone known locus-specific primer is firstly used to enrich the targeted template DNA strand resulting in significant PCR product specificity during the second round conventional nested PCR. The distance between the known locus-specific primer and the nearest location of the restriction enzyme used determined the length of the obtained PCR product. We used this technique to walk downstream into the isochorismatase and upstream into the hypothetical conserved genes flanking the mature extracellular lipase gene from Bacillus licheniformis. We further demonstrated the potential of the technique as a cost-effective method during PCR-based prospecting for novel genes by designing "universal" degenerate primers that detected homologues of Family VII bacterial lipolytic genes in Bacillus species. The cassette ligation-mediated PCR was used to clone complete nucleotide sequences encoding functional lipolytic genes from B. licheniformis and Bacillus pumilus. PMID:15722149

  18. Investigation of spore surface antigens in the genus Bacillus by the use of polyclonal antibodies in immunofluorescence tests.

    PubMed

    Phillips, A P; Martin, K L

    1988-01-01

    Fluorescein-conjugated rabbit antibodies to formalized spores of Bacillus anthracis were tested against strains of B. anthracis and other Bacillus species in a subjective immunofluorescence test. The lack of reaction of B. anthracis Vollum spores with conjugated antibody raised against B. anthracis Sterne spores indicated that spores of the Vollum strain lacked a major surface antigen present in most of the other anthrax strains tested, including the non-encapsulated strains Sterne and the Soviet ST1, variants cured of the pX01 plasmid that codes for the toxin, and several virulent strains. Four other antibody preparations, raised against B. anthracis Vollum, New Hampshire, Ames and Strain 15, reacted to an approximately similar degree with spores of all four strains and of Sterne, indicating that Vollum has at least one spore antigen in common with these other strains. The anti-Sterne and anti-Vollum conjugates both displayed cross-reactions with spores of strains of B. cereus, B. coagulans, B. subtilis, B. megaterium, B. polymyxa, B. pumilus and B. thuringiensis. Absorption of the anti-anthrax conjugates with B. cereus NCTC 8035 and NCTC 10320 removed all these cross-reactions, demonstrating the existence of spore antigens specific for anthrax. PMID:3127370

  19. Genetic diversity and characterization of heavy metal-resistant-endophytic bacteria from two copper-tolerant plant species on copper mine wasteland.

    PubMed

    Sun, Le-Ni; Zhang, Yan-Feng; He, Lin-Yan; Chen, Zhao-Jin; Wang, Qing-Ya; Qian, Meng; Sheng, Xia-Fang

    2010-01-01

    The diversity of endophytic bacteria from the copper-tolerant species Elsholtzia splendens and Commelina communis was evaluated by using cultivation and cultivation-independent techniques. Phylogenetic analysis based on 16S rDNA sequences showed that the Cu-resistant isolates belonged to three major phylogenetic groups: Firmicutes, Actinobacteria and Proteobacteria. Bacillus and Acinetobacter were predominant among the Cu-resistant isolates. Sequence analysis from the 16S rDNA clone libraries of the two plant roots revealed sequences of Alpha-, Beta-, Gamma-Proteobacteria, Firmicutes,Actinobacteria and Bacteroidetes. Gammaproteobacteria was predominant in the two 16S rDNA clone libraries. Thirty-two endophytic bacteria were isolated and characterized with respect to heavy metal resistance and production of plant growth-promoting factors. In experiments involving rape plants grown in vermiculite containing 4 mg kg(-1) of Cu, inoculation with the isolates was found to increase the dry weights of roots (ranging from 132% to 155%) and above-ground tissues (ranging from 71% to 83%) compared to the uninoculated control. Increase in above-ground tissue Cu content varied from 63% to 125% in inoculated-rape plants cultivated in Cu-contaminated substrate compared to the uninoculated control. PMID:19762232

  20. Isolation and characterization of endophytic plant growth-promoting bacteria from date palm tree (Phoenix dactylifera L.) and their potential role in salinity tolerance.

    PubMed

    Yaish, Mahmoud W; Antony, Irin; Glick, Bernard R

    2015-06-01

    Endophytic bacteria were isolated from date palm (Phoenix dactylifera L.) seedling roots, characterized and tested for their ability to help plants grow under saline conditions. Molecular characterization showed that the majority of these strains belonged to the genera Bacillus and Enterobacter and had different degrees of resistance to various antibiotics. Some of these strains were able to produce the enzyme 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase and the plant growth regulatory hormone indole-3-acetic acid (IAA). Some strains were also able to chelate ferric iron (Fe(3+)) and solubilize potassium (K(+)), phosphorus (PO 4 (3-) ) and zinc (Zn(2+)), and produce ammonia. The results also showed that ACC deaminase activity and IAA production was slightly increased in some strains in response to an increase in NaCl concentration in the growth media. Consistent with these results, selected strains such as PD-R6 (Paenibacillus xylanexedens) and PD-P6 (Enterobacter cloacae) were able to enhance canola root elongation when grown under normal and saline conditions as demonstrated by a gnotobiotic root elongation assay. These results suggest that the isolated and characterized endophytic bacteria can alter ethylene and IAA levels and also facilitate nutrient uptake in roots and therefore have the potential role to promote the growth and development of date palm trees growing under salinity stress. PMID:25860542