Struktur Und Dynamik Heterotropher Bakteriengemeinschaften Im Wattenmeer Und Der Deutschen Bucht

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Struktur Und Dynamik Heterotropher Bakteriengemeinschaften Im Wattenmeer Und Der Deutschen Bucht Struktur und Dynamik heterotropher Bakteriengemeinschaften im Wattenmeer und der Deutschen Bucht Structure and dynamics of heterotrophic bacterial communities in the German Wadden Sea and the German Bight Dissertation zur Erlangung des akademischen Grades einer Doktorin der Naturwissenschaften (Dr. rer. nat.) der Fakultät V Mathematik und Naturwissenschaften der Carl von Ossietzky Universität Oldenburg vorgelegt von Beate Rink geboren am 23.01.1974 in Bremerhaven Erstgutachter : Prof. Dr. Meinhard Simon Zweitgutachter: Prof. Dr. Heribert Cypionka Eingereicht am: Disputation am: Für Rosemarie Erklärung Teilergebnisse dieser Arbeit sind als Beiträge bei den genannten Fachzeitschriften eingereicht oder werden eingereicht. Mein Beitrag an der Erstellung der verschiedenen Manuskripte wird im Folgenden erläutert: Rink, B. , Seeberger, S., Martens, T., Duerselen, C. D., Simon, M., und Brinkhoff, T. (2006) Effects of a phytoplankton bloom in a coastal ecosystem on the composition of bacterial communities (Eingereicht bei Aquat. Microb. Ecol. ) Etablierung und Spezifitätstest der Roseobacter spezifischen PCR durch S. S. unter Anleitung von B. R. und T. B (Diplomarbeit, 2003). Durchführung der spezifischen PCR und DGGE, der Klonierung und Sequenzierung durch B. R. Statistische Auswertung und Erstellung der phylogenetischen Stammbäume durch B. R. Erstellung der ersten Fassung des Manuskripts durch B. R., Überarbeitung durch T. B., B. R. und M. S. Rink, B., Martens, T., Fischer, D., Lemke, A., Grossart, H. P., Simon, M., und Brinkhoff, T. (2006) Tidal effects on coastal bacterioplankton (In Vorbereitung zum Einreichen bei Limnol. Oceanogr. ) Planung und Durchführung der Probenahme 2005 durch B. R. Durchführung der spezifischen PCR und DGGE sowie der RNA Untersuchungen und CARD-FISH durch B. R. Statistische Auswertung und Erstellung der phylogenetischen Stammbäume durch B. R. Erstellung der ersten Fassung des Manuskripts durch B. R., Überarbeitung durch T. B., B. R. und M. S. Stevens, H., Brinkhoff, T., Rink, B. , Vollmers, J., und Simon, M. (2006) Diversity and abundance of Gram-positive bacteria in a tidal flat ecosystem (Eingereicht bei Environ. Microbiol. ) Durchführung der spezifischen CARD-FISH und DGGE Untersuchungen von J. V. unter Anleitung von B. R. und T. B (Leistungsnachweis, 2005). Überarbeitung des Manuskriptes von B. R., T. B. und M. S. Rink, B. , Brinkhoff, T., Ziegelmüller, K., und Simon, M. (2006) High regional variability of bacterial communities in the German Bight, North Sea (Eingereicht bei Aquat. Microb. Ecol. ) Planung und Durchführung der Probenahme 2002 von Mirko Lunau und B. R. Planung und Durchführung der Probenahme 2003 von B. R. Molekularbiologische Untersuchungen (PCR, DGGE, Klonierung, Sequenzierung), statistische Auswertung und Erstellung der phylogenetischen Stammbäume durch B. R. Erstellung der ersten Fassung des Manuskripts durch B. R., Überarbeitung durch T. B., B. R. und M. S. Tagungsbeiträge Rink B , Stevens H, Simon M, Brinkhoff T (2006) Stability of Microbial Communities Within Different Time Scales in a Tidal Flat Ecosystem. Posterbeitrag, International Symposium Microbial Ecology (ISME-11), Wien, Österreich, 20-25 August Rink B , Brinkhoff T, Simon M (2004) Bacterial communities reflect different regional properties of the German Bight. Vortrag, VAAM-Meeting Braunschweig, 28-31 March Rink B , Kruse M, Seeberger S, Stevens H, Brinkhoff T, Simon M (2004) Seasonal and spatial differences in the composition and abundance of bacterial communities in the German Bight of the North Sea. Posterbeitrag, International Symposium Microbial Ecology (ISME-10), Cancun, Mexico, 22-27 August Simon M, Selje N, Schledjewski R, Rink B , Grossart HP (2004) Diversity and substrate turnover of bacterioplankton communities in the Gulf of Aqaba, Red Sea. Posterbeitrag, International Symposium Microbial Ecology (ISME-10), Cancun, Mexico, 22-27 August Rink B , Lunau M, Seeberger S, Stevens H, Brinkhoff T, Grossart H-P, Simon M (2003) Diversity patterns of aggregate-associated and free-living bacterial communities in the German Wadden Sea. In Rullkötter J. (ed.), BioGeoChemistry of Tidal Flats - Proceedings of a Workshop held at the Hanse Institute of Advanced Study, Delmenhorst (Germany), 14- 17 May. Forschungszentrum Terramare, Wilhelmshaven, Berichte Nr. 12, 96-98. ISSN 1432-797X. Lunau M, Rink B , Grossart H-P, Simon M (2003) How to sample marine microaggregates in shallow and turbid environments? - Problems and solutions. In Rullkötter J. (ed.), BioGeoChemistry of Tidal Flats - Proceedings of a Workshop held at the Hanse Institute of Advanced Study, Delmenhorst (Germany), 14-17 May. Forschungszentrum Terramare, Wilhelmshaven, Berichte Nr. 12, 85-88. ISSN 1432-797X. Rink B , Brinkhoff T, Simon M (2002) Completing the picture of natural habitats: The use of specific Primersets in DGGE. Posterbeitrag, VAAM Meeting, Berlin, 23-26 March Zusammenfassung Im Wattenmeer unterliegen die Organismen hochdynamischen Prozessen. Eine flache Wassersäule und der Einfluß der Gezeiten sorgen für starke Strömungen und hohe Resuspensionsraten. Auch der tidale Ein- und Ausstrom von Wassermassen aus der Nordsee in das Rückseitenwatt beeinflusst das System. Während in Herbst- und Wintermonaten sedimentologische Faktoren überwiegen, ist im Frühjahr und Sommer ein deutlicher Einfluss biologischer Größen nachweisbar. Im Rahmen des interdisziplinären Forschungsprojekts „Biogeochemie des Watts“, in das diese Arbeit eingebunden ist, wurden große Varianzen innerhalb des Schwebstoffaufkommens sowie in bakterieller Aktivität und Abundanz auf saisonaler Ebene sowie im Tidenzyklus beschrieben. In der vorliegenden Arbeit wurde untersucht, inwiefern tidale und saisonale Faktoren die Struktur der ansässigen Bakteriengemeinschaften in der Wassersäule beeinflussen. Weiterführend wurde untersucht, ob die im Wattenmeer detektierten Phylotypen standortspezifisch oder auch in anderen Gebieten der Deutschen Bucht nachweisbar sind. Im Wattenmeer fand die Beprobung in der Otzumer Balje im Rückseitenwatt von Spiekeroog statt. Im ersten Teil dieser Arbeit wurden zur Untersuchung des Zusammenhangs von Bakteriengemeinschaften und Phytoplankton wöchentlich Proben genommen und mittels gruppenspezifischer DGGE (Denaturierende Gradienten Gelelektrophorese) und statistischer Methoden untersucht. Im zweiten Teil wurden neben saisonalen auch tidale Vorgänge beleuchtet. Die Probenahme fand im Herbst, Frühjahr und Sommer in einstündigem und dreistündigem Probenahmeraster statt. Die Bakteriengemeinschaften wurden mittels gruppenspezifischer DGGE für alpha-Proteobakterien , Bacteroidetes und Roseobacter sowohl DNA- als auch RNA basiert untersucht. Zusätzlich wurden FISH (Fluoreszens in situ Hybridisierung) und die hoch sensitive CARD-FISH (Catalyzed Reporter Deposition-FISH) eingesetzt und somit erstmalig die Abundanzen einzelner Bakteriengruppen in der Wassersäule des Wattenmeeres dargestellt. In einer vorangegangenen Arbeit wurden im Wattenmeer bemerkenswert viele gram-positive Bakterien isoliert, was zu der Annahme führte, dass diese Bakteriengruppe eine besondere Stellung in diesem Habitat einnimmt. Zur Vervollständigung der Daten wurde im dritten Teil dieser Arbeit die CARD-FISH eine Actinobakterien -spezifische Sonde eingesetzt und zusätzlich eine spezifische DGGE entwickelt, um Abundanz und phylogenetische Vielfalt der Actinobakterien im Watt zu untersuchen. Die Probenahme hierzu wurde an verschiedenen Standorten im Spiekerooger Rückseitenwatt durchgeführt. Im vierten Teil wurden im Sommer 2002 und 2003 verschiedene Standorte der Deutschen Bucht an der Küstenzone, vor Helgoland und in der offenen Nordsee beprobt. Die Bakteriengemeinschaften wurden mit spezifischer DGGE für alpha-Proteobakterien und Bacteroidetes untersucht. Zur weiteren Beschreibung der Ökologie an den untersuchten Standorten wurden zusätzlich hydrologische, mikrobiologische und partikuläre Parameter bestimmt. Zusammenfassend ergaben sich aus diesen Arbeiten folgende Hauptaussagen: • Im Wattenmeer sind die Bakteriengemeinschaften in der Wassersäule im Wesentlichen aus alpha- und gamma-Proteobakterien sowie Bacteroidetes zusammengesetzt. Darüber hinaus sind beta-Proteobakterien abundant auf Aggregaten. Hierbei bilden frei lebende und Aggregat-assoziierte Bakterien distinkte Gemeinschaften sowohl im Wattenmeer als auch in der Deutschen Bucht. Die Struktur der frei lebender Bakteriengemeinschaften besteht hauptsächlich aus wenigen dominanten Phylotypen der Roseobacter Gruppe. Ihre Zusammensetzung ist saisonal und räumlich stabil. Die Struktur der Aggregat-assoziierten Bakterien zeigt grössere Artenvielfalt als bei frei lebenden Bakterien und unterliegt deutlicher räumlich- zeitlichen Einflussfaktoren. Hier dominieren Phylotypen innerhalb der Bacteroidetes , gamma- und delta-Proteobakterien . • Saisonale Einflüsse auf die Bakteriengemeinschaften sind in den produktiven Frühjahrs- und Sommermonaten erkennbar. Insbesondere Aggregat-assoziierte Bakterien der Roseobacter -Gruppe und Bacteroidetes unterliegen biologischen Einflussfaktoren wie Phytoplanktonblüten. Tidale Einflüsse auf bakterielle Aktivität und Abundanz werden nur geringfügig und nicht systematisch durch Änderungen in der Zusammensetzung der Bakteriengemeinschaften reflektiert. • Actinobakterien stellen knapp 5% des Bakterioplanktons im Wattenmeer. Ihre Abundanz und Zusammensetzung im Süßwasserbereich unterscheidet sich von den marinen Standorten, wobei frei lebende und Aggregat-assoziierte Actinobakterien distinkte Gemeinschaften bilden. Aus
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