Glycerol ether lipids in sediments: sources, diversity and implications Dissertation zur Erlangung des Doktorgrades der Naturwissenschaften Dr. rer. nat. Am Fachbereich Geowissenschaften Der Universität Bremen vorgelegt von Xiao-Lei Liu Bremen Mai 2011 Supervisor: Prof. Dr. Kai-Uwe Hinrichs (MARUM, University of Bremen, Germany) Co-supervisor: Prof. Dr. Roger E. Summons (EAPS, Massachusetts Institute of Technology, USA) Date of colloquium: 27 May, 2011 This study was funded by Deutsche Forschungsgemeinschaft (DFG, Germany) through the international graduate college EUROPROX for a scholarship to Xiao-Lei Liu. Works and thesis were finished between November 2007 and May 2011 within the Organic Geochemistry Group of the MARUM (Center for Marine Environmental Sciences) and Department of Geosciences, University of Bremen, Leobener Str., D-28359 Bremen, Germany. ‘I know that nothing good lives in me, that is, in my sinful nature.’ --- Romans 7:18 Table of Contents Abastract..........................................................................................................................I Zusammenfassung ..................................................................................................... III List of Figures ............................................................................................................VII List of Tables ............................................................................................................... IX Chapter 1. Introduction .................................................................................................. 1 1. Molecular proxies..................................................................................... 1 2. Deep biosphere in marine subsurface sediments: another source of organic molecule .......................................................................................... 4 3. Glycerol ethers as distinctive membrane lipids........................................ 6 4. Glycerol tetraethers in marine sediment................................................... 8 5. Methods.................................................................................................. 11 6. Objectives and thesis outline.................................................................. 12 7. Contributions to publications ................................................................. 14 Reference.................................................................................................... 17 Chapter 2. Identification of polar lipid precursors of the ubiquitous branched GDGT orphan lipids in a peat bog in Northern Germany ...................................... 23 Chapter 3. Distribution of intact and core GDGTs in marine sediments..................... 43 Chapter 4. Isoprenoid glycerol dibiphytanol diethers: a series of novel archaeal lipids in marine sediments.................................................................................... 65 Chapter 5. Novel mono- and dihydroxy-glycerol dibiphytanyl glycerol tetraethers and their intact polar analogues in marine sediments ....................................... 81 Chapter 6. Distribution of glycerol ether lipids in marine sediment: structural diversity and biogeological implications................................................................. 105 Chapter 7. Conclusions and perspectives................................................................... 137 1. Conclusions .......................................................................................... 137 2. Perspectives.......................................................................................... 140 Reference.................................................................................................. 143 Acknowledgements.................................................................................................. 145 Abstract Abstract Glycerol ether lipids are prominent membrane constituents in Archaea and Bacteria that are characterized by high potential for preservation in geological settings. During the past decade they were increasingly used in molecular proxies. For example, selected glycerol dialkyl glycerol tetraethers (GDGT) are used in ratios such as the TEX86 and BIT index for reconstructing past sea surface temperature (SST) and terrestrial input, respectively. However, the distribution and structural diversity of glycerol ether lipids in marine sediments has not yet been fully explored. In order to obtain a better understanding of the origin and fate of these lipids and to evaluate the potential impact on molecular proxies, a globally distributed set of samples was analyzed in this PhD thesis project. More than forty novel compounds were revealed and it could be shown that the diversity of glycerol ether lipids in marine sediments is much higher than previously recognized. Among the studied lipids, isoprenoid GDGTs were shown as the most dominant component in all analyzed sediments with over 70% of total ether lipids. A comparison of the lipid composition of core and intact polar isoprenoid GDGTs pointed to a potential impact of live benthic archaea and their intact lipids on the application of the TEX86 SST proxy. The same study suggested that recycling of fossil GDGTs from planktonic archaea by the benthic archaeal community could be an important process. In a set of peat sediments, two intact polar lipids of the orphan branched GDGTs were for the first time observed. These branched GDGTs that constitute the BIT index have in their intact form glucuronosyl and glucosyl headgroups. The two compounds accounted for 4-7% of total IPLs, suggesting that their producers represent a sizeable but not dominant component of the microbial community. The identification of these intact polar precursors of these orphan lipids provides important constraints for the search of their microbial sources. The presence of newly identified glycerol ether lipids with distinctive structures and ubiquitous distribution in all analyzed marine sediments provided potential phylogenetic biomarkers and a large reservoir for novel molecular proxies to be developed. A series of novel ether core lipids coexisting with corresponding isoprenoid GDGTs was identified in all analyzed sediments. The first series of compounds, I Zusammenfassung accounting on average for 7% of total ether lipids, was identified as glycerol dibiphytanol diethers (GDDs) and is considered to represent either biosynthetic intermediates or degradation products of GDGTs. A second series was identified as hydroxylated GDGTs based on nuclear magnetic resonance (NMR) and mass spectral interpretation. Accordingly, a series of unknown IPLs that had been previously reported as a major component in samples of archaeal cultures, sediments, and the water column were then recognized to be glycosidic hydroxy-GDGTs. The widespread occurrence of IPLs of hydroxy-GDGTs suggests an important contribution of specific archaeal species with high activity in a wide range of geological settings. Several other groups of novel compounds were also tentatively identified based on interpretation of mass spectra. Specifically, extended H-shaped GDGTs, hybrid isoprenoid/branched GDGTs, and overly and sparsely branched GDGTs are characterized with a common feature of methylation series, containing compounds with one -CH2- unit difference between each other. It is expected that the information encoded in the distribution of the novel lipids can be used to develop molecular proxies indicating past environmental factors such as temperature and salinity and/or information pertaining to the composition and activity of extant microbial communities. An overall distribution of detected glycerol ether lipids in marine subsurface sediments was shown by the estimated relative abundance of 11 structural groups, and therefore reflects a general composition of ether lipid producing microbes contributing to the sedimentary record. II Abstract Zusammenfassung Glyzerolbasierte Etherlipide sind bedeutende Membranbestandteile von Archaeen und Bakterien und zeichnen sich durch ein hohes Erhaltungspotential im geologischen Kontext aus. In den letzten zehn Jahren erhielten sie eine zunehmende Bedeutung für molekulare Proxies. Bestimmte Glyzerol-dialkyl-glyzerol-tetraetherlipide (GDGT) kommen z.B. zur Anwendung im TEX86 und BIT Index welche zur Rekonstruktion von Oberflächenwassertemperaturen beziehungsweise terrestrischem Eintrag in Sedimente in der Vergangenheit benutzt werden können. Allerdings ist die Verteilung und strukturelle Vielfalt von glyzerolbasierten Etherlipiden in marinen Sedimenten immernoch nicht vollständig erforscht. Um einen besseren Überblick über die Herkunft und das Schicksal dieser Lipide zu bekommen und um einen potentiellen Einfluss auf molekulare Proxies abschätzen zu können, wurde in der vorliegenden Doktorarbeit ein global verteilter Probensatz analysiert. Mehr als vierzig neue Komponenten wurden entdeckt und es konnte gezeigt werden dass die Diversität von glyzerolbasierten Etherlipiden in marinen Sedimenten sehr viel höher ist als bisher angenommen. Den grössten Anteil an den erforschten Lipiden haben die isoprenoiden GDGTs, welche die dominante Komponente mit einem relativen Anteil von mehr als 70% von allen Etherlipiden ausmachten. Ein Vergleich der Lipidzusammensetzung von GDGT Kernlipiden und intakten polaren isoprenoiden GDGTs deutete auf einen potentiellen Einfluss von lebenden benthischen Archaeen und deren intakten Lipiden auf den TEX86 Oberflächenwassertemperatur Proxy