Antimicrobial Properties of Pseudomonas Strains Producing
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Evolutionary Patchwork of an Insecticidal Toxin
Ruffner et al. BMC Genomics (2015) 16:609 DOI 10.1186/s12864-015-1763-2 RESEARCH ARTICLE Open Access Evolutionary patchwork of an insecticidal toxin shared between plant-associated pseudomonads and the insect pathogens Photorhabdus and Xenorhabdus Beat Ruffner1, Maria Péchy-Tarr2, Monica Höfte3, Guido Bloemberg4, Jürg Grunder5, Christoph Keel2* and Monika Maurhofer1* Abstract Background: Root-colonizing fluorescent pseudomonads are known for their excellent abilities to protect plants against soil-borne fungal pathogens. Some of these bacteria produce an insecticidal toxin (Fit) suggesting that they may exploit insect hosts as a secondary niche. However, the ecological relevance of insect toxicity and the mechanisms driving the evolution of toxin production remain puzzling. Results: Screening a large collection of plant-associated pseudomonads for insecticidal activity and presence of the Fit toxin revealed that Fit is highly indicative of insecticidal activity and predicts that Pseudomonas protegens and P. chlororaphis are exclusive Fit producers. A comparative evolutionary analysis of Fit toxin-producing Pseudomonas including the insect-pathogenic bacteria Photorhabdus and Xenorhadus, which produce the Fit related Mcf toxin, showed that fit genes are part of a dynamic genomic region with substantial presence/absence polymorphism and local variation in GC base composition. The patchy distribution and phylogenetic incongruence of fit genes indicate that the Fit cluster evolved via horizontal transfer, followed by functional integration of vertically transmitted genes, generating a unique Pseudomonas-specific insect toxin cluster. Conclusions: Our findings suggest that multiple independent evolutionary events led to formation of at least three versions of the Mcf/Fit toxin highlighting the dynamic nature of insect toxin evolution. -
Pseudomonas Chlororaphis PA23 Biocontrol of Sclerotinia Sclerotiorum on Canola
Pseudomonas chlororaphis PA23 Biocontrol of Sclerotinia sclerotiorum on Canola: Understanding Populations and Enhancing Inoculation LORI M. REIMER A Thesis Submitted to the Faculty of Graduate Studies In Partial Fulfillment of the Requirements For the Degree of MASTER OF SCIENCE Department of Plant Science University of Manitoba Winnipeg, Manitoba © Copyright by Lori M. Reimer 2016 GENERAL ABSTRACT Reimer, Lori M. M.Sc., The University of Manitoba, June 2016. Pseudomonas chlroraphis PA23 Biocontrol of Sclerotinia sclerotiorum on Canola: Understanding Populations and Enhancing Inoculation. Supervisor, Dr. Dilantha Fernando. Pseudomonas chlororaphis strain PA23 has demonstrated biocontrol of Sclerotinia sclerotiorum (Lib.) de Bary, a fungal pathogen of canola (Brassica napus L.). This biocontrol is mediated through the production of secondary metabolites, of which the antibiotics pyrrolnitrin and phenazine are major contributers. The objectives of this research were two-fold: to optimize PA23 phyllosphere biocontrol and to investigate PA23’s influence in the rhizosphere. PA23 demonstrated longevity, both in terms of S. sclerotiorum biocontrol and by having viable cells after 7 days, when inoculated on B. napus under greenhouse conditions. Carbon source differentially effected growth rate and antifungal metabolite production of PA23 in culture. PA23 grew fasted in glucose and glycerol, while mannose lead to the greatest inhibition of S. sclerotiorum mycelia and fructose lead to the highest levels of antibiotic production relative to cell density. Carbon source did not have a significant effect on in vivo biocontrol. PA23 demonstrated biocontrol ability of the fungal root pathogens Rhizoctonia solani J.G. Kühn and Pythium ultimum Trow in radial diffusion assays. PA23’s ability to promote seedling root growth was demonstrated in sterile growth pouches, but in a soil system these results were reversed. -
Management of Tomato Diseases Caused by Fusarium Oxysporum
Crop Protection 73 (2015) 78e92 Contents lists available at ScienceDirect Crop Protection journal homepage: www.elsevier.com/locate/cropro Management of tomato diseases caused by Fusarium oxysporum R.J. McGovern a, b, * a Chiang Mai University, Department of Entomology and Plant Pathology, Chiang Mai 50200, Thailand b NBD Research Co., Ltd., 91/2 Rathburana Rd., Lampang 52000, Thailand article info abstract Article history: Fusarium wilt (FW) and Fusarium crown and root rot (FCRR) of tomato (Solanum lycopersicum) caused by Received 12 November 2014 Fusarium oxysporum f. sp. lycopersici and F. oxysporum f. sp. radicis-lycopersici, respectively, continue to Received in revised form present major challenges for production of this important crop world-wide. Intensive research has led to 21 February 2015 an increased understanding of these diseases and their management. Recent research on the manage- Accepted 23 February 2015 ment of FW and FCRR has focused on diverse individual strategies and their integration including host Available online 12 March 2015 resistance, and chemical, biological and physical control. © 2015 Elsevier Ltd. All rights reserved. Keywords: Fusarium oxysporum f. sp. lycopersici F. oxysporum f. sp. radicis-lycopersici Fusarium wilt Fusarium crown and root rot Solanum lycopersicum Integrated disease management Host resistance Biological control Methyl bromide alternatives 1. Background production. Losses from FW can be very high given susceptible host- virulent pathogen combinations (Walker,1971); yield losses of up to Fusarium oxysporum represents a species complex that includes 45% were recently reported in India (Ramyabharathi et al., 2012). many important plant and human pathogens and toxigenic micro- Losses from FCRR in greenhouse tomato have been estimated at up organisms (Nelson et al., 1981; Laurence et al., 2014). -
Università Degli Studi Di Padova Dipartimento Di Biomedicina Comparata Ed Alimentazione
UNIVERSITÀ DEGLI STUDI DI PADOVA DIPARTIMENTO DI BIOMEDICINA COMPARATA ED ALIMENTAZIONE SCUOLA DI DOTTORATO IN SCIENZE VETERINARIE Curriculum Unico Ciclo XXVIII PhD Thesis INTO THE BLUE: Spoilage phenotypes of Pseudomonas fluorescens in food matrices Director of the School: Illustrious Professor Gianfranco Gabai Department of Comparative Biomedicine and Food Science Supervisor: Dr Barbara Cardazzo Department of Comparative Biomedicine and Food Science PhD Student: Andreani Nadia Andrea 1061930 Academic year 2015 To my family of origin and my family that is to be To my beloved uncle Piero Science needs freedom, and freedom presupposes responsibility… (Professor Gerhard Gottschalk, Göttingen, 30th September 2015, ProkaGENOMICS Conference) Table of Contents Table of Contents Table of Contents ..................................................................................................................... VII List of Tables............................................................................................................................. XI List of Illustrations ................................................................................................................ XIII ABSTRACT .............................................................................................................................. XV ESPOSIZIONE RIASSUNTIVA ............................................................................................ XVII ACKNOWLEDGEMENTS .................................................................................................... -
(12) United States Patent (10) Patent No.: US 7476,532 B2 Schneider Et Al
USOO7476532B2 (12) United States Patent (10) Patent No.: US 7476,532 B2 Schneider et al. (45) Date of Patent: Jan. 13, 2009 (54) MANNITOL INDUCED PROMOTER Makrides, S.C., "Strategies for achieving high-level expression of SYSTEMIS IN BACTERAL, HOST CELLS genes in Escherichia coli,” Microbiol. Rev. 60(3):512-538 (Sep. 1996). (75) Inventors: J. Carrie Schneider, San Diego, CA Sánchez-Romero, J., and De Lorenzo, V., "Genetic engineering of nonpathogenic Pseudomonas strains as biocatalysts for industrial (US); Bettina Rosner, San Diego, CA and environmental process.” in Manual of Industrial Microbiology (US) and Biotechnology, Demain, A, and Davies, J., eds. (ASM Press, Washington, D.C., 1999), pp. 460-474. (73) Assignee: Dow Global Technologies Inc., Schneider J.C., et al., “Auxotrophic markers pyrF and proC can Midland, MI (US) replace antibiotic markers on protein production plasmids in high cell-density Pseudomonas fluorescens fermentation.” Biotechnol. (*) Notice: Subject to any disclaimer, the term of this Prog., 21(2):343-8 (Mar.-Apr. 2005). patent is extended or adjusted under 35 Schweizer, H.P.. "Vectors to express foreign genes and techniques to U.S.C. 154(b) by 0 days. monitor gene expression in Pseudomonads. Curr: Opin. Biotechnol., 12(5):439-445 (Oct. 2001). (21) Appl. No.: 11/447,553 Slater, R., and Williams, R. “The expression of foreign DNA in bacteria.” in Molecular Biology and Biotechnology, Walker, J., and (22) Filed: Jun. 6, 2006 Rapley, R., eds. (The Royal Society of Chemistry, Cambridge, UK, 2000), pp. 125-154. (65) Prior Publication Data Stevens, R.C., “Design of high-throughput methods of protein pro duction for structural biology.” Structure, 8(9):R177-R185 (Sep. -
Classification of Isolates from the Pseudomonas Fluorescens
ORIGINAL RESEARCH published: 15 March 2017 doi: 10.3389/fmicb.2017.00413 Classification of Isolates from the Pseudomonas fluorescens Complex into Phylogenomic Groups Based in Group-Specific Markers Daniel Garrido-Sanz, Eva Arrebola, Francisco Martínez-Granero, Sonia García-Méndez, Candela Muriel, Esther Blanco-Romero, Marta Martín, Rafael Rivilla and Miguel Redondo-Nieto* Departamento de Biología, Facultad de Ciencias, Universidad Autónoma de Madrid, Madrid, Spain The Pseudomonas fluorescens complex of species includes plant-associated bacteria with potential biotechnological applications in agriculture and environmental protection. Many of these bacteria can promote plant growth by different means, including modification of plant hormonal balance and biocontrol. The P. fluorescens group is Edited by: currently divided into eight major subgroups in which these properties and many other Martha E. Trujillo, ecophysiological traits are phylogenetically distributed. Therefore, a rapid phylogroup University of Salamanca, Spain assignment for a particular isolate could be useful to simplify the screening of putative Reviewed by: Youn-Sig Kwak, inoculants. By using comparative genomics on 71 P. fluorescens genomes, we have Gyeongsang National University, identified nine markers which allow classification of any isolate into these eight South Korea subgroups, by a presence/absence PCR test. Nine primer pairs were developed for David Dowling, Institute of Technology Carlow, Ireland the amplification of these markers. The specificity and sensitivity of these primer pairs *Correspondence: were assessed on 28 field isolates, environmental samples from soil and rhizosphere and Miguel Redondo-Nieto tested by in silico PCR on 421 genomes. Phylogenomic analysis validated the results: [email protected] the PCR-based system for classification of P. -
Hydrogen Cyanide, Which Contributes to Pseudomonas Chlororaphis Strain PA23 Biocontrol, Is Upregulated in the Presence of Glycine ⇑ Munmun Nandi A, Carrie Selin B, G
Biological Control 108 (2017) 47–54 Contents lists available at ScienceDirect Biological Control journal homepage: www.elsevier.com/locate/ybcon Hydrogen cyanide, which contributes to Pseudomonas chlororaphis strain PA23 biocontrol, is upregulated in the presence of glycine ⇑ Munmun Nandi a, Carrie Selin b, G. Brawerman c, W.G. Dilantha Fernando b, Teresa de Kievit a, a Department of Microbiology, University of Manitoba, Winnipeg, MB R3T 2N2, Canada b Department of Plant Science, University of Manitoba, Winnipeg, MB R3T 2N2, Canada c Department of Pharmacology and Therapeutics, University of Manitoba, Winnipeg, MB R3T 2N2, Canada highlights graphical abstract HCN contributes to Pseudomonas chlororaphis PA23 biocontrol in vitro and planta. Glycine increases production of HCN, phenazine and pyrrolnitrin. Glycine upregulates the PhzRI quorum-sensing system. In quorum-sensing deficient strains, FeCl3 increases hcn gene expression. article info abstract Article history: Pseudomonas chlororaphis strain PA23 is a biocontrol agent capable of suppressing the pathogenic fungus Received 5 December 2016 Sclerotinia sclerotiorum. This bacterium secretes the antibiotics pyrrolnitrin (PRN) and phenazine (PHZ), Revised 10 February 2017 together with degradative enzymes and siderophores. Strain PA23 also produces hydrogen cyanide Accepted 21 February 2017 (HCN); however, the role of this compound in PA23 antifungal (AF) activity remains unknown. The Available online 24 February 2017 aim of the current study was to characterize an hcn mutant and determine whether HCN contributes to biocontrol. Analysis of an hcn mutant revealed decreased AF activity both in vitro and in a newly estab- Keywords: lished model of S. sclerotiorum root-rot infection in lettuce. When glycine (20 mM) and ferric chloride Antifungal (100 mM) were included as media amendments, elevated AF activity was observed. -
1 Molecular and Environmental Plant Sciences Texas A&M University
MEPS academic program review 2014 Molecular and Environmental Plant Sciences Texas A&M University External Review Molecular and Environmental Plant Sciences Graduate Program Self Study March 2014 Molecular and Environmental Plant Sciences 1 MEPS academic program review 2014 Molecular and Environmental Plant Sciences Texas A&M University Molecular and Environmental Plant Sciences Graduate Program Self Study March 2014 College of Agriculture and Life Sciences College of Geosciences College of Science Molecular and Environmental Plant Sciences 2 MEPS academic program review 2014 TABLE OF CONTENTS EXECUTIVE SUMMARY 5 1. INTRODUCTION 8 1.1 Welcome 1.2 Charge to the Review Team 2. TEXAS A&M UNIVERSITY SYSTEM 10 2.1 Texas A&M University 2.2 The College of Agriculture and Life Sciences (COALS) 2.3 Aggie Traditions 2.4 Graduate Interdisciplinary Programs 3. MEPS PROGRAM STRUCTURE 18 3.1 Program History 3.2 Program Description 3.3 University Administration of MEPS Program 3.4 Administrative Structure of the MEPS Program 3.4.1 Executive Committee and Chair 3.4.2 Program Coordinator 3.4.3 Admissions Committee 3.4.4 Symposium Committee 3.4.5 Nomination and Awards Committee 3.5 Budget Allocations and Program Expenditures 3.5.1 Funding for current graduate students 4. THE MEPS GRADUATE PROGRAM 31 4.1 Admission Requirements and Procedures 4.2 The Curriculum 4.2.1 Master of Molecular and Environmental Plant Sciences 4.2.2 Doctoral Degree 4.2.3 Degree Plan 4.2.4 Advisory Committee 4.2.5 Designated MEPS Courses 4.2.6 MEPS funded Assistantships 4.2.7 MEPS Graduate Student Engagement 5. -
Antibiotic Resistant Pseudomonas Spp. Spoilers in Fresh Dairy Products: an Underestimated Risk and the Control Strategies
foods Review Antibiotic Resistant Pseudomonas Spp. Spoilers in Fresh Dairy Products: An Underestimated Risk and the Control Strategies Laura Quintieri , Francesca Fanelli * and Leonardo Caputo Institute of Sciences of Food Production, National Research Council of Italy, Via G. Amendola 122/O, 70126 Bari, Italy * Correspondence: [email protected]; Tel.: +39-0805929317 Received: 19 July 2019; Accepted: 23 August 2019; Published: 1 September 2019 Abstract: Microbial multidrug resistance (MDR) is a growing threat to public health mostly because it makes the fight against microorganisms that cause lethal infections ever less effective. Thus, the surveillance on MDR microorganisms has recently been strengthened, taking into account the control of antibiotic abuse as well as the mechanisms underlying the transfer of antibiotic genes (ARGs) among microbiota naturally occurring in the environment. Indeed, ARGs are not only confined to pathogenic bacteria, whose diffusion in the clinical field has aroused serious concerns, but are widespread in saprophytic bacterial communities such as those dominating the food industry. In particular, fresh dairy products can be considered a reservoir of Pseudomonas spp. resistome, potentially transmittable to consumers. Milk and fresh dairy cheeses products represent one of a few “hubs” where commensal or opportunistic pseudomonads frequently cohabit together with food microbiota and hazard pathogens even across their manufacturing processes. Pseudomonas spp., widely studied for food spoilage effects, are instead underestimated for their possible impact on human health. Recent evidences have highlighted that non-pathogenic pseudomonads strains (P. fluorescens, P. putida) are associated with some human diseases, but are still poorly considered in comparison to the pathogen P. aeruginosa. -
Characterisation of Pseudomonas Spp. Isolated from Foods
07.QXD 9-03-2007 15:08 Pagina 39 Annals of Microbiology, 57 (1) 39-47 (2007) Characterisation of Pseudomonas spp. isolated from foods Laura FRANZETTI*, Mauro SCARPELLINI Dipartimento di Scienze e Tecnologie Alimentari e Microbiologiche, sezione Microbiologia Agraria Alimentare Ecologica, Università degli Studi di Milano, Via Celoria 2, 20133 Milano, Italy Received 30 June 2006 / Accepted 27 December 2006 Abstract - Putative Pseudomonas spp. (102 isolates) from different foods were first characterised by API 20NE and then tested for some enzymatic activities (lipase and lecithinase production, starch hydrolysis and proteolytic activity). However subsequent molecular tests did not always confirm the results obtained, thus highlighting the limits of API 20NE. Instead RFLP ITS1 and the sequencing of 16S rRNA gene grouped the isolates into 6 clusters: Pseudomonas fluorescens (cluster I), Pseudomonas fragi (cluster II and V) Pseudomonas migulae (cluster III), Pseudomonas aeruginosa (cluster IV) and Pseudomonas chicorii (cluster VI). The pectinolytic activity was typical of species isolated from vegetable products, especially Pseudomonas fluorescens. Instead Pseudomonas fragi, predominantly isolated from meat was characterised by proteolytic and lipolytic activities. Key words: Pseudomonas fluorescens, enzymatic activity, ITS1. INTRODUCTION the most frequently found species, however the species dis- tribution within the food ecosystem remains relatively The genus Pseudomonas is the most heterogeneous and unknown (Arnaut-Rollier et al., 1999). The principal micro- ecologically significant group of known bacteria, and bial population of many vegetables in the field consists of includes Gram-negative motile aerobic rods that are wide- species of the genus Pseudomonas, especially the fluores- spread throughout nature and characterised by elevated cent forms. -
Marta Dora Phd Thesis
MOLECULAR MONITORING OF MEAT SPOILING PSEUDOMONAS SPECIES AND ANALYSIS OF STAPHYLOCOCCAL ENTEROTOXIN EXPRESSION AND FORMATION Dóra Márta Doctoral Thesis Corvinus University of Budapest Faculty of Food Science Department of Microbiology and Biotechnology Budapest, 2011 ii According to the Doctoral Council of Life Sciences of Corvinus University of Budapest on 29 th November 2011, the following committee was designated for defence. Committee: Chair: Tibor Deák, D.Sc. Members: Péter Biacs, D.Sc. László Abrankó, Ph.D. Judit Tornai-Lehoczki, Ph.D. Judit Beczner, C.Sc. Opponents: Adrienn Micsinai, Ph.D. László Varga, Ph.D. Secretary: László Abrankó, Ph.D. iii “I am among those who think that science has great beauty. A scientist in his laboratory is not only a technichian: he is also a child placed before natural phenomena which impress him like a fairy tale.” Marie Curie (1867-1934) iv CONTENTS LIST OF ABBREVIATION............................................................................................................vii 1. INTRODUCTION..........................................................................................................................1 2. LITERATURE REVIEW..............................................................................................................3 2.1. Characterization of red meat.....................................................................................................3 2.1.1. Microbiological aspects of food spoilage especially on pork............................................4 2.2. Food -
Aquatic Microbial Ecology 80:15
The following supplement accompanies the article Isolates as models to study bacterial ecophysiology and biogeochemistry Åke Hagström*, Farooq Azam, Carlo Berg, Ulla Li Zweifel *Corresponding author: [email protected] Aquatic Microbial Ecology 80: 15–27 (2017) Supplementary Materials & Methods The bacteria characterized in this study were collected from sites at three different sea areas; the Northern Baltic Sea (63°30’N, 19°48’E), Northwest Mediterranean Sea (43°41'N, 7°19'E) and Southern California Bight (32°53'N, 117°15'W). Seawater was spread onto Zobell agar plates or marine agar plates (DIFCO) and incubated at in situ temperature. Colonies were picked and plate- purified before being frozen in liquid medium with 20% glycerol. The collection represents aerobic heterotrophic bacteria from pelagic waters. Bacteria were grown in media according to their physiological needs of salinity. Isolates from the Baltic Sea were grown on Zobell media (ZoBELL, 1941) (800 ml filtered seawater from the Baltic, 200 ml Milli-Q water, 5g Bacto-peptone, 1g Bacto-yeast extract). Isolates from the Mediterranean Sea and the Southern California Bight were grown on marine agar or marine broth (DIFCO laboratories). The optimal temperature for growth was determined by growing each isolate in 4ml of appropriate media at 5, 10, 15, 20, 25, 30, 35, 40, 45 and 50o C with gentle shaking. Growth was measured by an increase in absorbance at 550nm. Statistical analyses The influence of temperature, geographical origin and taxonomic affiliation on growth rates was assessed by a two-way analysis of variance (ANOVA) in R (http://www.r-project.org/) and the “car” package.