Improved Taxonomy of the Genus Streptomyces

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Improved Taxonomy of the Genus Streptomyces UNIVERSITEIT GENT Faculteit Wetenschappen Vakgroep Biochemie, Fysiologie & Microbiologie Laboratorium voor Microbiologie Improved taxonomy of the genus Streptomyces Benjamin LANOOT Scriptie voorgelegd tot het behalen van de graad van Doctor in de Wetenschappen (Biochemie) Promotor: Prof. Dr. ir. J. Swings Co-promotor: Dr. M. Vancanneyt Academiejaar 2004-2005 FACULTY OF SCIENCES ____________________________________________________________ DEPARTMENT OF BIOCHEMISTRY, PHYSIOLOGY AND MICROBIOLOGY UNIVERSITEIT LABORATORY OF MICROBIOLOGY GENT IMPROVED TAXONOMY OF THE GENUS STREPTOMYCES DISSERTATION Submitted in fulfilment of the requirements for the degree of Doctor (Ph D) in Sciences, Biochemistry December 2004 Benjamin LANOOT Promotor: Prof. Dr. ir. J. SWINGS Co-promotor: Dr. M. VANCANNEYT 1: Aerial mycelium of a Streptomyces sp. © Michel Cavatta, Academy de Lyon, France 1 2 2: Streptomyces coelicolor colonies © John Innes Centre 3: Blue haloes surrounding Streptomyces coelicolor colonies are secreted 3 4 actinorhodin (an antibiotic) © John Innes Centre 4: Antibiotic droplet secreted by Streptomyces coelicolor © John Innes Centre PhD thesis, Faculty of Sciences, Ghent University, Ghent, Belgium. Publicly defended in Ghent, December 9th, 2004. Examination Commission PROF. DR. J. VAN BEEUMEN (ACTING CHAIRMAN) Faculty of Sciences, University of Ghent PROF. DR. IR. J. SWINGS (PROMOTOR) Faculty of Sciences, University of Ghent DR. M. VANCANNEYT (CO-PROMOTOR) Faculty of Sciences, University of Ghent PROF. DR. M. GOODFELLOW Department of Agricultural & Environmental Science University of Newcastle, UK PROF. Z. LIU Institute of Microbiology Chinese Academy of Sciences, Beijing, P.R. China DR. D. LABEDA United States Department of Agriculture National Center for Agricultural Utilization Research Peoria, IL, USA PROF. DR. R.M. KROPPENSTEDT Deutsche Sammlung von Mikroorganismen & Zellkulturen (DSMZ) Braunschweig, Germany DR. F. MARINELLI Vicuron Pharmaceuticals, Italy PROF. DR. F. GOSSELE Visiting Professor in Microbiology Phibro Animal Health, Belgium DR. G. HUYS Faculty of Sciences, University of Ghent PROF. DR. E. VANDAMME Faculty of Applied Biological Sciences, University of Ghent 5 6 Dankwoord - Acknowledgements Hierbij wens ik een aantal personen in het zonnetje te plaatsen die elk hun steentje hebben bijgedragen tot het succesvol beëindigen van deze thesis. Tijdens mijn studentenjaren maakten Moniek G en Joris M me wegwijs in de wondere wereld van de bacteriën in het practicum Microbiologie en konden me overtuigen op de WE10V-trein te stappen voor mijn licentiaatsthesis. Een samenwerking met het International Rice Research Institute te Manilla (Filippijnen) deed me echter finaal kiezen voor doctoraal onderzoek. Dankzij Karel K, Danny J, Philippe D en Marc V werd een Belgo-Chinese samenwerking afgesloten voor initieel 2 jaar in 1998, dat 2 maal verlengd zou worden met 2 jaar. Moniek D van het Federaal Wetenschapsbeleid zorgde voor de financiering van dit project. Zonder jullie inzet en (federale) centen zou dit boekje er niet geweest zijn ! Jean, je gaf me als promotor de nodige ruimte om me te verdiepen in dit onderzoek. Ik wil je tevens bedanken voor je eindeloze steun en vertrouwen, de talloze kritische opmerkingen en verbeteringen. Kortom je was de noodzakelijke, krachtige locomotief die de zware “Streptomyces-project trein” gedurende 6 jaar door soms krappe bochten sleurde naar zijn finale bestemming. Marc V, je was een uitstekend co-promotor die me van begin tot einde telkenmale overstelpte met inspirerende inzichten, soms zeer ontnuchterende maar terechte opmerkingen (krijg ik het slachtbankje mee naar huis?) maar die steeds voldoende energie kon overbrengen om niet de moed op te geven. Zonder jouw hulp zou dit werk moeilijk verlopen zijn. Hartelijk dank hiervoor Marc ! Verder wil ik ook alle collega’s van het Laboratorium voor Microbiologie inclusief Moudjahid, Maja (mijn chaperon), Medana, Pavel en Teresa bedanken voor de aangename werksfeer. In het bijzonder wil ik mijn “Streptomyces-reisgezellen” Margo, Ann VS, en de thesisstudentes Annick P, Nancy en Lieve bedanken voor hun inspanningen bij het screenen van de honderden isolaten en referentiestammen. Karen L, Cindy, Fabiano, Renata, Liesje, Ilse C, Evie en An B voor hun raad bij DNA-bereidingen, DNA-DNA hybridisaties en % GC bepalingen; Vinie en Anne W voor de hulp tijdens de tDNA-PCR experimenten, Roberto om het Italiaanse temperament binnengebracht te hebben in ons labo; Antoine voor het oplossen van de technische probleempjes; Paul S voor de vele schouderklopjes en de leuke antiquiteiten; Urbain voor je opbeurende gesprekken en sympathie; Stefanie V voor het frivole pasta-diner (...) tijdens een congres in Parijs; Dirk D voor je uitstekende hulp bij eiwitprofilering, me de knepen van de fascinerende fotografiewereld bijgebracht te hebben en je levenswijsheid; Annemie en “baron” Fernand voor de administratieve steun; snelle Jeanine voor het leveren van allerhande producten; Peter D voor de geapprecieerde hulp bij de numerieke verwerking van de massa’s data en Geert K voor het oplossen van de onvermijdelijke computerproblemen, zijn talloze mopjes en het mede sponsoren van de jaarlijks georganiseerde mattetaartenslag (het waren echte Geraardsbergse !). Last but not least dien ik de collega’s van het LMG zelf te bedanken voor het verzamelen van de referentiestammen (dank u Claudine), lyofiliseren, etc van de toch moeilijk handelbare streptomyceten. Er zijn er ongetwijfeld die grijze haren gekregen hebben... This work was performed in collaboration with the Institute of Microbiology of the Chinese Academy of Sciences in Beijing (P.R. China). I would like to thank Prof Liu Zhiheng, Li Wei, Jianli Zhang, Liming Wang, Huang Ying and Miss Zhang to help constructing the reference frameworks, the isolation of hundreds of strains from different soil samples 7 collected around China and to communicate all their knowledge on traditional streptomycete systematics which is needed to understand properly the classification of these soil bacteria. Besides the professional side, I will not forget the hospitality shown by the whole group, the many nice dinners, the efforts made to get a European boy acquainted with the fascinating old Chinese culture and to always provide me with a guide when visiting Beijing… It has been an unforgettable experience despite the sometimes-encountered language frontier. I am also very grateful to the group of Michael Goodfellow (Newcastle, UK) in helping to interpret our data and how these might fit the overall picture. The members of the examination committee are acknowledged for their critical reading and comments. Deel van dit werk kwam tot stand dmv een samenwerking met de industrie namelijk PHIBRO Animal Health te Rixensart. In het bijzonder Francis en Martine wens ik te bedanken voor hun kritische commentaren en inzichten. Bedankt beste ouders, broertje en zusje voor jullie onvoorwaardelijke steun en de nodige ontspanning die jullie boden. Dit is werkelijk onbetaalbaar ! Ten slotte zijn er nog de de enthousiaste supporters Marie-Jeanne VDB (Europacollege), Hans S, Jean Pierre L, Geert M, Rudy DB, Arlette DD en het personeel van FGRA. Heidi, kusjes om me steeds op de sporen gehouden te hebben tijdens de laatste maanden van het schrijven. Gent, 9 december 2004 8 CONTENTS Dankwoord - Acknowledgements Contents List of non-standard abbreviations List of Figures List of Tables PART 1: Literature review Chapter 1: The genus Streptomyces 23 1.1 Introduction 1.2 Circumscription 1.3 Phylogenetic allocation 1.4 Cultivation 1.5 Occurrence 1.6 Importance 1.6.1 Human pathogens 1.6.2 Plant pathogens 1.6.3 Humification and biodegradation 1.6.4 Biodeterioration 1.6.5 Bioremediation 1.6.6 Production of enzymes 1.6.7 Production of natural products 1.6.8 Bioconversion of chemical compounds Chapter 2: Taxonomy of the genus Streptomyces 51 2.1 Introduction 2.2 The early chaotic years 2.3 Numerical taxonomy 2.4 Chemotaxonomy 2.5 DNA-DNA hybridisations 2.6 Genotyping screening methods 2.7 Phylogeny 2.7.1 16S rDNA sequence analysis 2.7.2 Analysis of other stretches DNA 2.7.2.1 Sequencing of protein coding genes 2.7.2.2 16S rDNA – 23S rDNA ITS analysis 2.7.3 Whole genome sequencing and analysis Chapter 3: Aims & Conceptual framework 85 3.1 General aims 3.2 Specific aims 9 PART 2: Experimental work Chapter 4: Detection and reclassification of synonymous species 91 4.1 The search for synonyms among streptomycetes using SDS-PAGE of whole-cell proteins. Emendation of the species Streptomyces aurantiacus, Streptomyces cacaoi subsp. cacaoi, Streptomyces caeruleus and Streptomyces violatus. Lanoot B, Vancanneyt M, Cleenwerck I, Wang L, Li W, Liu Z & Swings J. (2002). Int J Syst Evol Microbiol 52, 823-829. 4.2 BOX-PCR fingerprinting as a powerful tool to reveal synonymous names in the genus Streptomyces. Emended descriptions are proposed for the species Streptomyces cinereorectus, S. fradiae, S. tricolor, S. colombiensis, S. filamentosus, S. vinaceus and S. phaeopurpureus. Lanoot B, Vancanneyt M, Dawyndt P, Cnockaert MC, Zhang J, Huang Y, Liu Z & Swings J. (2004). System Appl Microbiol 27, 84-92. 4.3 Reclassification of Streptomyces nigrifaciens as a later synonym of Streptomyces flavovirens; Streptomyces citreofluorescens, Streptomyces chrysomallus subsp. chrysomallus and Streptomyces fluorescens as later synonyms of Streptomyces anulatus; Streptomyces chibaensis as later synonym of Streptomyces corchorusii; Streptomyces flaviscleroticus as later synonym of S. minutiscleroticus;
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